(Advances in Parasitology 31) J.R. Baker and R. Muller (Eds.) - Academic Press (1992)

Editorial Board
C. Bryant Department of Zoology, Australian National University, G.P.O.

Box 4,Canberra, A.C.T. 2601, Australia
C. Combes Laboratoire de Biologie Animale, UniversitC de Perpignan,

Avenue de Villeneuve, 66860 Perpignan Cedex, France
M. Coluzzi Director, Instituto di Parassitologia, Universita Delgi Studi di

Roma “La Sapienza”, P. le A. Moro 5 , 00185 Roma, Italy
W.H.R. Lumsden 17A Merchiston Crescent, Edinburgh EHlO 5AX, UK
E.J.L. Soulsby Department of Clinical Veterinary Medicine, University of

Cambridge, Madingley Road, Cambridge CB3 OES, UK
K.Tanabe Laboratory of Biology, Osaka Institute of Technology, Ohmiya,

Asahi-Ku, Osaka 535, Japan
K.S. Warren Comprehensive Medical Systems, Inc., 46 1 Fifth Avenue,

New York, N.Y. 10017, USA
P. Wenk Tropenmedizinisches Institut, Universitat Tubingen, D7400

Tubingen 1, Wilhelmstrasse 3 1, Germany
Advances in
PARASITOLOGY
Edited by
J.R. BAKER
Royal Society of Tropical Medicine and Hygiene,
London, England
R. MULLER
International Institute of Parasitology,
St Albans, England
and
D. ROLLINSON
The Natural History Museum,
London, England
VOLUME 35
ACADEMIC PRESS
Harcourt Brace & Company, Publishers
London San Diego New York Boston
Sydney Tokyo Toronto
ACADEMIC PRESS LIMITED
24/28 Oval Road
LONDON NWI 7DX
United States Edition published by

ACADEMIC PRESS INC.
San Diego CA 92101
Copyright 0 1995, by
ACADEMIC PRESS LIMITED
This book is printed on acid-free paper
All Rights Reserved

No part of this book may be reproduced in any form by photostat,
microfilm, or any other means, without wiitten permission
from the publishers
A catalogue record for this book is available from the British Library
ISBN 0-12-031735-4
Typeset by J&L Composition Ltd, Filey, North Yorkshire

Printed in Great Britain by T.J. Press (Padstow) Ltd, Padstow, Comwall
F. ATHIAS-BINCHE, Laboratoire Arago, Universitk Paris VIICNRS UA I 17,
,' 66650 Banyuls sur Mer, France
W.C. CAMPBELL, The Charles A. Dana Research Institute for Scientists
Emeriti, Drew University, Madison, New Jersey 07940, USA
G.A. CONDER,Upjohn Laboratories, The Upjohn Company, Kalamazoo,
Michigan 490014199, USA
C.C. CONSTANTINE, WHO Collaborating Centre for the Molecular
Epidemiology of Parasitic Infections and Institute for Molecular
Genetics and Animal Disease, School of Veterinary Studies, Murdoch
University, Western Australia 6150, Australia
M . DE JONG-BRINK, Graduate School of Neurosciences Amsterdam,
Research Institute of Neurosciences Vrije Universiteit, Faculty of
Biology, Vrije Universiteit, De Boelelaan 1087, 1081 HV Amsterdam,
The Netherlands
M . HALL,Department of Entomology, The Natural History Museum,
Cromwell Road, London SW7 5BD, UK
t M . KALISZEWSKI, Brigham Young University, Provo, Utah 84602, USA
E.E. LINDQUIST, Centre for Land and Biological Resources Research,
Agriculture Canada, K. W. Neatby Building, CEF, Ottawa, Ontario
KIA OC6, Canada
A.J. LYMBERY, WHO Collaborating Centre for the Molecular Epidemiology
of Parasitic Infections and Institute for Molecular Genetics and Animal
Disease, Western Australian Department of Agriculture, Bunbury,
Western Australia 6230, Australia
K. MACKENZIE, SOAFD Marine Laboratory, PO Box 101, Victoria Road,
Aberdeen AB9 8DB, UK
A.H. MCVICAR, SOAFD Marine Laboratory, PO Box 101, Victoria Road,
Aberdeen AB9 8DB, UK
R. SIDDALL,Department of Biology, University of Jyvaskyla, Seminaarinkatu
15, SF-40100 Jyvaskyla, Finland
R.C.A. THOMPSON, WHO Collaborating Centre for the Molecular
Epidemiology of Parasitic Infections and Institute for Molecular
Deceased.
V
vi CONTRIBUTORS TO VOLUME 35
Genetics and Animal Disease, School of Veterinary Studies, Murdoch

University, Western Australia 61.50, Australia
R. WALL,School of Biological Sciences, University of Bristol, Bristol BS8
I TH, UK
B. WILLIAMS, Department of Zoology, The National Museum of Wales,
Cathays Park, Cardiff, CFI 3NP, UK
H.H. WILLIAMS, Department of Zoology, The National Museum of Wales,
Cathays Park, Cardiff CFI 3NP and School of Pure and Applied
Biology, University of Wales College of Cardiff, Cardiff CFI 3TL, UK
The volume opens with an authoritative and comprehensive review of
cliemotherapy of nematode infections of veterinary importance by George
Conder of Upjohn Laboratories and William Campbell of Drew University.
Currently available drugs which have been approved for use against
nematode parasites in various animal species are considered. Particular
attention is given to the macrocyclic lactones, including ivermectin, the
first of these anthelmintics, as well as abamectin, moxidectin, milbemycin
B-41D, milbemycin oxime and doramectin. Drug resistance is an increas-
ingly important problem and is often encountered in the treatment of
domestic animals. This aspect of chemotherapy is examined in detail
with emphasis being given to the experience gained from the three classes
of modem broad-spectrum anthelmintics. Resistance is an important issue
but unfortunately is generally not recognized until it becomes a problem.
The authors provide a considered view concerning the impact and spread of
resistance, the factors influencing the spread, mechanisms of resistance,
monitoring of resistance and strategies that could be adopted to limit
resistance.
Ken MacKenzie from the SOAFD Marine Laboratory, Aberdeen and his
colleagues draw our attention to a possible useful role for parasites of fish.
They examine the potential use of parasites, especially helminths, as
indicators of water quality. Three specific categories of pollution -
hydrocarbon, heavy metal and thermal - are known to influence the
numbers and distribution of marine parasites and a detailed account of
the effects of each form of pollution is given. Examples are also provided
of the ways in which pollution may influence populations of fish parasites
via the invertebrate intermediate hosts and non-piscine vertebrate hosts.
The relationship between pollution and marine parasite ecology is clearly
complex and the influence of other biotic and abiotic factors must be
understood before pollution can be shown to be the cause of population
changes. The free-living transmission stages of helminth parasites may be
sensitive to environmental change and monitoring of appropriate species
may prove to be a measure of water quality. Guidelines and procedures for
the selection of the most appropriate host-parasite systems are detailed.
vii
viii PREFACE
Clearly, if parasites are to be useful as indicators of water quality a sound

knowledge of their biology and ecology will be a necessary prerequisite.
Hydatid disease (echinococcosis) is one of the most important parasitic
zoonoses and yet there has been a great deal of uncertainty concerning the
taxonomic status and relationships of various species and strains. A sound
taxonomic framework is desirable in order to define wild and domestic
cycles of transmission and those parasites responsible for disease. There
are currently four recognized species of Echinococcus but the phylogenetic
relationships among species and strains are far from clear. Andrew
Thompson (Murdoch University) and his colleagues review the substantial
body of work that has been carried out concerned with elucidating the
’ identity and relationships within the genus. Primarily due to the application
of molecular characterization methods, new data have been generated over
the last six years which are not in total agreement with the current sub-
generic classification. In this review the authors outline how a taxonomic
revision of the genus should proceed. As their starting point they consider
how a species of Echinococcus might be defined and develop their ideas by
considering the various species and strains. The authors put forward seven
species and suggest that detailed comparative studies should now be
undertaken in a number of endemic areas in order to determine the
geographic distribution and uniformity of the species so that formal
taxonomic designations can be confirmed.
An exciting and somewhat neglected area of research concerns the
interaction between the immunological response of an organism and its
neuroendocrine system as well as the physiological effects resulting from
such interactions. Experimental studies on host-parasite interactions tend
to focus on the interplay between either the immune system and the
parasite or the neuroendocrine system and the parasite but rarely on path-
ways linking the two regulatory systems. In this comprehensive review
Marijke De Jong-Brink (Vrije University) provides a fascinating insight
into such interactions by considering the schistosome-host model. Careful
experimentation with Trichobilharzia ocellata and its snail host Lymnaea
stagnalis has shown how the parasite affects both reproduction and growth
of its snail host. The important role of the humoral factor schistosomin is
considered in detail. The review explores the wider issue of whether it is a
general survival strategy for parasites to profit from the stress responses
they evoke in their host and provides an impetus for further research on
other host-parasite interactions.
Martin Hall of the Natural History Museum, London and Richard Wall
of the University of Bristol have fully reviewed recent advances in medical
and veterinary myiasis. Undoubtedly the most momentous event in this
field in recent years has been the accidental introduction of the American
screwworm, Cochliomyia hominivorax, into Libya in 1988 with potentially
PREFACE ix
catastrophic effects if it reached countries south of the Sahara. Fortunately,

after a determined (and expensive) campaign described by the authors, the
fly appears to have been eradicated. In recent years there has been increas-
ing emphasis on quantitative analytical studies rather than the traditional
descriptive, case orientated, accounts and the authors provide a very useful
and fascinating resume of new monitoring, modelling and forecasting
methods.
The volume ends with an account by Marek Kaliszewski (Brigham
Young University, Provo, Utah), Franqoise Athias-Binche (Universitd
Paris VI, Banyuls sur Mer) and Evert Lindquist (Centre for Land and
Biological Research, Agriculture Canada) of what is probably a rather
little-known group, the tarsonemine mites. These very interesting small
animals (less than OSmm long) are all associated in one way or another
with other arthropods or with vertebrates. The authors trace the evolu-
tionary.development within the group from simple phoresy to parasitoidism
and (by a separate route) to true parasitism, and consider the possibility that
the curious phenomenon of physogastry - massive enlargement of the
female’s body to permit the simultaneous development of a large number
of embryos - may be a preadaptation to parasitoidism.
Sadly, the first author of this paper (Dr Marek Kaliszewski) died as a
result of a car accident in October 1992, during the initial stages of
planning the paper. The third author very kindly agreed to take over his
role in its completion. The paper, therefore, represents a truly international
collaboration between scientists from the USA, France and Canada, and is
dedicated to the memory of Dr Kaliszewski.
JOHN BAKER
RALPH MULLER
DAVID ROLLINSON
Chemotherapy of Nematode Infections of
Veterinary Importance. with Special Reference
to Drug Resistance
George A Conder.
Upjohn Laboratories. The Upjohn Company. Kalamazoo. Michigan. USA
and
.
William C Campbell
The Charles A . Dana Research Institute for Scientists Emeriti. Drew

University. Madison. New Jersey. USA
1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
2. TheDrugs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
3. New Methods of Drug Delivery . . . . . . . . . . . . . . . . . . . . . . 4
4. Macrocyclic Lactones . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
4.1. Mechanism of Action . . . . . . . . . . . . . . . . . . . . . . . . . 5
4.2. lvermectin .............................. 7
4.3. Abamectin .............................. 14
4.4. Moxidectin .............................. 15
4.5. Milbemycin B-41D .......................... 19
4.6. Milbemycin oxime .......................... 19
4.7. Doramectin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20
5. Prospective Drugs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22
5.1. Paraherquamide ........................... 22
5.2. PF1022A ............................... 25
5.3. Dioxapyrrolomycin .......................... 25
5.4. Clonostachydiol ............................ 26
. ....................
6 Resistance to Antinematodal Drugs 26
6.1. Is resistance an issue? ...
..................... 26
....................
6.2. Extent of resistance worldwide 29
ADVANCES IN PARASITOLOOY VOL 35 Copyright Q 1995 Academic Press Limited

ISBN 0-12-0317354 AN rights ofreprodurtion in any form resewed
2 GEORGE A. CONDER AND WILLIAM C. CAMPBELL
.
6.3. Causes of treatment failure . . . . . . . .............. 38
6.4. Continuing spread of resistance . . . . .
. .............. 42
6.5. Factors contributing-to resistance . . . .
. .............. 43
6.6. Mechanisms of resistance ......... .............. 45
6.7. Monitoringheportingof resistance . ... .............. 49
6.8. Strategies to limit resistance development .............. 54
6.9. Epilogue ................ ............... 57
Acknowledgements .............................. 57
Note added in proof .............. ............... 57
References . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . 58
+ 1. INTRODUCTION
The present time seems especially appropriate for a review of antinematode

chemotherapy, because we have entered what can only be described as “a
new era”. It is an era characterized by drugs more effective than ever
before and by treatment failures more menacing than ever before.
In the 1980s the macrocyclic lactone class of drugs was introduced to the
anthelmintic armamentarium. Their impact has been enormous for two
reasons. First, they quickly dominated the scene, reaching significant
proportions of the world population of livestock and companion animals
(to say nothing of capturing an unprecedented segment of the animal health
market). Second, because these drugs were effective against many ecto-
parasitic arthropods, they caused a fusion of the chemotherapy of nematode
and ectoparasite infections. Because of this, they have been somewhat
infelicitously characterized as “endectocides”. It should be borne in
mind that efficacy against acarines and insects is a factor in the demarca-
tion of the macrocyclic lactones as a category.of drugs quite distinct from
those of the past (such demarcation does not necessarily imply superiority,
of course, but acknowledges the succession to power of a new ruling class).
In this chapter, we limit our consideration of resistance to the realm of
nematodes; but we have thought it necessary to include the arthropod
parasites in our consideration of the efficacy of the newer drugs.
The historical succession of one drug class after another has resulted in
part from sequential improvement in drug characteristics (spectrum,
safety, convenience, etc.), but also in part from the emergence of drug
resistance. The phenomenon of resistance, so characteristic of the treatment
of infectious diseases in general, is a prominent feature of the new era in
antinematodal therapy, and we here devote major attention to it. Because of
the space devoted to this topic and to the macrocyclic drugs, there is no
discussion of other aspects such as the economics of treatment or the
methods by which anthelmintic drugs are discovered and evaluated. We
CHEMOTHERAPY OF NEMATODE INFECTIONS OF VETERINARY IMPORTANCE 3
give short shrift to older drugs and to new drug candidates that have not
reached the marketplace. We would emphasize, however, that whereas
some of the old standbys have been dropped from regular use others still
play important roles in the control of nematodiases.
2. THE DRUGS
Anthelmintic use may vary to some extent in various parts of the world, but
it :rests largely on a common set of chemical compounds. It may be
instructive, therefore, to list the compounds currently approved by the
United States Government for use against nematode parasites in various
animal species:
In beef cattle: albendazole, coumaphos, fenbendazole, haloxon, ivermectin,
levamisole, morantel tartrate, oxfendazole, phenothiazine, thiabendazole.
In non-lactating dairy cattle: albendazole, fenbendazole, haloxon,
ivermectin, levamisole, oxfendazole, phenothiazine, thiabendazole.
In lactating dairy cattle: coumaphos, morantel tartrate, thiabendazole.
In sheep: ivermectin, levamisole, phenothiazine, thiabendazole.
In goats: phenothiazine, thiabendazole.
In swine: dichlorvos, fenbendazole, hygromycin B, ivermectin, levamisole,
piperazine, pyrantel tartrate, thiabendazole.
In horses: butonate, cambendazole, dichlorvos, febantel, fenbendazole,
ivermectin, levamisole, mebendazole, oxfendazole, oxibendazole, pheno-
thiazine, piperazine, pyrantel pamoate, pyrantel tartrate, thiabendazole,
tioxidazole, trichlorfon.
In dogs: arsenamide sodium, butamisole hydrochloride, dichlorvos,
diethylcarbamazine citrate, disophenol sodium, diathiazanine iodide, feban-
tel, fenbendazole, glycobiarsol, ivermectin, mebendazole, milbemycin
oxime, oxibendazole, piperazine, pyrantel pamoate, styrylpyridinium
chloride, thenium closylate, ticarbodine, toluene.
In cats: dichlorvos, diethylcarbamazine citrate, disophenol sodium,
febantel, piperazine, toluene.
A few of these compounds (haloxon, butonate, cambendazole, butami-
sole) are not currently marketed, but remain approved (Courtney and
Sundlof, 1991).
Compounds approved by other regulatory agencies are likely to be the
same as those listed above or of the same chemical class. Anthelmintics vary
widely in efficacy, safety, and cost; selection of a compound for a particular
use is usually governed by such factors. Decisions on the strategy of
treatment (frequency and timing) will also depend on the epidemiological
circumstances. As discussed below, known or anticipated drug resistance

may also affect treatment decisions.
Apart from the macrocyclic lactones, the compounds listed on page 3 (and
others of their kind) will not be discussed in detail because they are so well
known. The older agents have been thoroughly documented (Bard, 1972;
Gibson, 1975). More recent reviews, in English, include the following:
Use in all major species: Blair and Klei (1986), Campbell (1986), Marriner
and Armour (1986), Raether (1988), Courtney and Sundlof (1991).
Strategy of use in all major species: Michel (1985).
Use in cattle, sheep, and goats: Boersema,(l985a), Coles (1986a), Prichard
(1986).
Strategy of use in cattle: Williams et al. (1986).
Strategy of use in sheep and goats: Craig (1986), Herd (1986d), Wescott
(1986a).
Chemistry, pharmacology, and mode of action: Fisher (1986), Rew and
Fetterer (1986), Vanden Bossche (1985).
Use in horses: Mirck (1989, Wescott (1 986b).
Use in swine: Biehl (1986), Rochette (1985a).
Use in dogs: Rochette (1985b).
Use in birds: Boersema (1985b).
Benzimidazoles: this class of anthelmintics contains so many derivatives
and is so widely used that reviews of this group have recently been
published side-by-side: Campbell (1990), Cook (1990), Denham (1990),
Gottschall et al. (1990), Horton (1990), Lacey (1990), Roos (1990),
Townsend and Wise (1990), Waller (1990).
3. NEW METHODS OF DRUG DELIVERY
An aspect of the older drugs that warrants brief consideration here is the
development of new delivery technology. As livestock production becomes
more intensified, labor-saving methods are eagerly sought. Devices have
been developed to provide sustained chemoprophylaxis or periodic
chemotherapy without the need for repeated and costly handling of animals.
The incorporation of anthelmintics into salt “licks”, molasses
“blocks”, and the like, is an old technique for getting livestock to ingest
drug over a prolonged period. Studies are being done on the application of
fenbendazole (one of the newer benzimidazoles) in this way (Miller et al.,
1992).
Morantel has been incorporated into two sustained-release devices for
cattle. The first was a ruminal bolus designed to provide delivery of drug
for 60-90 days. The second, more recent, device is a flexible plastic sheet
consisting of three layers, with morantel tartrate incorporated in the middle

layer. When the rolled-up sheet is administered orally, by balling-gun, the
sheet opens up in the rumen-reticulum and releases drug over a period of
about 90 days.
An albendazole device is administered to sheep, by balling-gun, and is
held in the rumen-reticulum by plastic “wings” that open out when it
reaches that destination. Over the next three months, a spring moves a
succession of drug tablets into place so that dissolution of drug can occur at
the surface of an exposed tablet. A device for cattle releases albendazole
three times at monthly intervals. Tiny batteries power an electronic system
that controls the gas-driven release of drug at the prescribed intervals.
Another device for cattle releases oxfendazole into the rumen-reticulum
at intervals of about three weeks over a period of approximately four
months. The device contains a magnesium alloy rod with spaced annular
tablets of drug. Continuous galvanic erosion of the rod in the rumen fluid
exposes each tablet in turn to provide pulsatile release of drug.
The great potency of the macrocyclic lactones has encouraged the
development of novel systems of delivery, and these will be described in
the section on these drugs.
4. MACROCYCLIC LACTONES
4.1. Mechanism of Action
Although the mechanism of action for the macrocyclic lactones is not fully
understood, it appears these compounds exert their effect by irreversibly
opening chloride channels in muscle membranes (Martin and Pennington,
1989). Contrary to early reports (reviewed by Turner and Schaeffer, 1989)
suggesting these chloride channels were associated with y-aminobutyric
acid (GABA) receptors, more recent evidence (reviewed by Geary et al.,
1992b) seems to indicate that there is no GABA association. Arena el al.
(1991, 1992) have proposed that the anthelmintic activity of the macro-
cyclic lactones is mediated by an interaction with a glutamate-gated
chloride channel; this conclusion is based on electrophysiological examina-
tion of membrane currents recorded from Xenopus laevis oocytes injected
with Caenorhabditis elegans RNA. It is unclear where receptors relevant
for anthelmintic activity for the macrocyclic lactones are located. Geary et
al. (1993) speculate that altered pharynx function (e.g. nutrient ingestion,
excretion, or regulation of turgor pressure) may be the actual site of
anthelmintic action as opposed to somatic musculature function, since
ivermectin inhibits pharyngeal pumping more potently than motility in
Haemonchus contortus. A similar pattern of ivermectin sensitivity in

pharynx versus somatic muscle has been demonstrated for Trichostrongylus
colubriformis (Bottjer arid Bone, 1985; Thomas as reported in Geary et al.,
1993). Further, laser ablation of all neurons in the circumesophageal
ganglion of C. elegans does not block pharyngeal pumping action, which
suggests the macrocyclic lactone receptors are located on the muscle
membrane, as opposed to neurons that innervate it.
It is likely that all macrocyclic lactone anthelmintics share the same
mechanism of action, although most of the work outlined above was done
using ivermectin. As shown in Figure 1, ivermectin, milbemycin D, and
moxidectin induce qualitatively similar ionotropic effects on membrane
'
conductance (in a shore crab muscle fiber preparation described by
Bowman et al., 1991b) which were reversed by subsequent addition of
picrotoxinin, a potent blocker of chloride channels. These and similar data
for several other macrocyclic lactones (Bowman et al., 1991b) seem to
support the notion of a common mechanism of action for these compounds.
An early report (Calcott and Fatig, 1984) suggested the activity of
V V
0 I I I I I 1 1
Figure 1 Time-dependent effects of 0.1 p~ ivennectin (0).avermectin B1 (m),

milbemycin D (o),and moxidectin (0)on shore crab muscle membrane resistance.
The anthelmintics or 0.1% dimethyl sulphoxide vehicle alone (A) were added at 0
time (first arrow). After 15 min (second arrow), the medium was replaced with
medium containing 10 p~ picrotoxinin, a C1-channel blocker. Data points are the
means 2 se of the percentage change in membrane resistance (Rm)recorded in at
least three separate preparations. (Data are courtesy of Dr David P. Thompson.)
macrocyclic lactones against nematodes was due to inhibition of chitin

metabolism, based on studies done with a fungus, Mucor miehei, and the
brine shrimp, Artemia salina. This possibility was later proven to be invalid
when it was demonstrated avermectin B1, of commercial grade did not
inhibit chitin metabolism of fungi (Onishi and Miller, 1985) or insects
(Gordnier et al., 1987). Similar grade milbemycin D also had no effect on
chitin metabolism (Gordnier et al., 1987). Further, Onishi and Miller
(1985) demonstrated an avermectin sample prepared in an identical
manner to that used by Calcott and Fatig (1984) contained oligomycin
and a polyene which accounted for the effect on chitin metabolism.
4.2. Ivermectin
Ivermectin is the 22, 23-dihydro derivative of the natural product aver-

mectin B;, and its structure is shown in Figure 2. Avermectin B1 is
produced by the filamentous bacterium Streptomyces avermitilis. Because
ivermectin was the first of the new wave of macrocyclic lactone anthel-
mintics, many hundreds of papers have been written about it, and it has
been reviewed many times (e.g. Campbell et al., 1983; Campbell and Benz,
1984; Barragry, 1987; Baker and Swain, 1989; Campbell, 1989; Goa et al.,
1991).
4.2.1. General Properties

Ivermectin is an off-white powder, largely insoluble in water but very
soluble in organic solvents. With the use of detergents, micellar aqueous
solutions can be prepared, and are in commercial use.
Ivermectin has a high therapeutic index, not because of extraordinary
safety in mammals (in absolute terms) but because of extraordinary
potency against parasites. In rats and mice, the acute LDS0 is in the range
25-50 mg kg-I. In dogs, signs of acute toxicity are seen first at 2.5 mg
kg-I, are significant at 5 mg kg-’, and are severe at 10 mg kg-I. In
rodents, it is fetotoxic only at dosages that approximate the maternotoxic
dosage. Fetotoxicity was not seen in extensive trials done in target species
for registration purposes or in women who were treated during early
pregnancy. There is now an extensive record of the safety of ivermectin
when given to millions of domestic animals at dosages of 200-500 pg kg-’
and to millions of humans at dosages of about 150 pg kg-’. Idiosyncratic
toxicity is well known to occur in some dogs (especially long-haired collie
breeds) but not at dosages used in commerce. Because ivermectin is lethal
to many arthropods, including “good” and “bad” dung-breeding insects,
concern has been expressed about interference with the natural biodegra-
dation of dung on pastures and consequent adverse environmental impact.
R R A B X
J J
cn CH
x) Yw
n
Figure 2 Structures of macrocyclic lactones.
Ivermectin is absorbed when given orally or parenterally, the rate of

absorption depending on the route of administration and the formulation
used. With the injectable formulation for cattle, for example, the plasma
peak is reached in two days, but experimental formulations can give peaks
in as little as one day. When given orally to dogs, horses, or humans, the
plasma peak is reached after 2-5 h (longer in horses given a paste formula-
tion). Most of the administered ivermectin is excreted in the feces of
treated animals as intact drug. The drug is lipophilic; in cattle slaughtered
seven days after treatment, residues found in liver, bile, and fat were far
higher than in other tissues.
4.2.2. Antiparasitic Utility

The following formulations are approved for commercial use: the trade-
marks cited are those of Merck & Co., Inc., except for Zimecterin, which is
registered by Farnam Co., Inc.
(a) Cattle
1. IVOMEC Injection: sterile solution containing 1% ivermectin, to be
injected subcutaneously at a dosage of 200 pg ivermectin kg-’.
2. IVOMEC F Injection: a sterile solution as above, but containing suffi-
cient clorsulon to give a dosage of 2 mg clorsulon kg-’ for control of
Fasciola hepatica.
3. IVOMEC Oral Liquid: solution containing 0.4% ivermectin, to be
administered so as to give a dosage of 200 pg ivermectin kg-’.
4. IVOMEC Oral Paste: a paste containing 0.153% ivermectin, delivered
by special applicator so as to give a dosage of 200 pg ivermectin kg-’.
5. IVOMEC Pour-on Liquid: a liquid (often, but inaccurately, called a
“topical” formulation) containing 0.5% ivermectin, to be applied to
the hide in a narrow strip along the back of the animal. The recom-
mended application rate of 1 ml 10 kg-’ body weight will deliver a
dosage of 500 pg ivermectin kg-’.
6 . IVOMEC Sustained Release Bolus: a bolus designed to lodge in the
rumen-reticulum of calves following administration by balling-gun.
Ivermectin is released continuously at a rate of 12 mg day-’ for a
period of 135 days, and the bolus is thus intended to provide parasite
control for an entire grazing season in the Northern Hemisphere markets
to which it has been introduced.
Administration of these formulations results in efficacy against the
immature and mature stages of many nematodes, and these are listed in
Table 1*. Susceptible parasites include gastrointestinal and pulmonary
species, levamisole/pyrantel/morantel-and benzimidazole-resistant strains,
* The efficacy tables presented herein on the macrocyclic lactones list only those parasites
for which an “approved claim” has been granted by a regulatory agency of high standing in
the animal health field (examples include the official governmental agencies of the United
States of America, the United Kingdom, Brazil, South Africa, Australia, New Zealand).
There are several advantages to this approach. (1) Approval is based on data from multiple
trials and on data that are considered persuasive by experts other than the original investi-
gators or the developers of the drug. (2) The registration dossiers include safety data; and so,
regulatory approvals offer more practical information than, say, a scientific publication that
records striking efficacy against a particular parasite at a dosage that turns out to be only
marginally safe. (3) The official registrations are based on marketed formulations (or
formulations shown experimentally to be bioequivalent). In contrast, a given scientific
publication may record data obtained with a formulation that differs from marketed products
in pharmacokinetic properties, and thus, may differ in safety and efficacy. The discrepancy
may or may not be noted in the journal article, but the problem does not arise in the context
of claims approved for a particular product.
Table 1 Parasites of cattle for which the use of ivermectin has received governmental approval. Where treatment is cited as "aids
in the control of" that too reflects regulatory approval. Approvals are listed for oral and injectable formulations only; for efficacy of
cutaneous "pour-on" formulation, see text.
Gastrointestinal Other nematodes Grubs Lice Mites Ticks
nematodes
Haemonchus placei (adultab,L? Dictyocaulus Hypoderma bovis Damalinia Chorioptes bovisk Boophilus
4") viviparus (lst, 2nd & 3rd bovisbC microplusk
(adult & L4)"b instars)ab
Ostertagia ostertagi (adultab, Parafilaria Hypoderma Linognathus Psoroptes ovisb Boopbilus
L? & L 4 a ) bovicolab lineatum vituliab (Psoroptes decoloratusk
(1st, 2nd & 3rd communis)
instars)"b
Ostertagia lyrata (adult & L4)"b Thelazia spp. Dermatobia Haematopinus Sarcoptes scabieib Ornithodoros
(adulgb hominis" eurysternusab savignyib
Solenopotes
capillatusb
Trichostrongylus axei (adult & L4)ab
Trichostrongylus colubriformis
(adult & L4)"b
Cooperia oncophora (adult & L )"w
Cooperia punctata (adult & 4 ) "%
Cooperia pectinata (adult & L<b
Nematodirus helvetianis (adult" & L4")
Nematodirus spathiger (adult)b
Strongyloides papillosus (adult)"b
Oesophagostomum radiatum (adult &
L4)ab
Bunostomum phlebotomum (adult, L3
Jz L4)b
Trichuris sp. (adult)b
" Oral formulation. Injectable formulation. Treatment aids in control. Including hypobiotic larvae.
and inhibited (hypobiotic) stages. An unexpected feature of the efficacy

profile is the “persistent” effect, which appears to derive from the rela-
tively prolonged drug absorption associated with certain formulations. For
some period after treatment of cattle with these formulations, newly
ingested nematode larvae fail to become established. As a result of this
activity, the injectable product has approved claims for the control of
Osrertagia sp. and Cooperia spp. for up to 7 days after treatment of the
host, and Dictyocaulus viviparus for up to 14 days. The cutaneous “pour-
on” formulation has approved claims for the control of Ostertagia sp. for
up to 14 days, and Dictyocaulus viviparus for up to 28 days, after treatment
of the host. Efficacy is also achieved against a variety of arthropod parasites,
including not only the “ectoparasitic” stages of insects and acarines, but
also the internal stages (grubs, warbles) of certain fly species (Table 1). In
general, parenteral administration (subcutaneous or cutaneous) gives better
activity against ectoparasites than does oral treatment.
The pour-on (cutaneous) formulation has an efficacy profile similar to
the other formulations, but with some differences. It is not fully effective
against the mite Psoroptes ovis, but has an approved claim for the control
of Chorioptes bovis. It is the only formulation with an approved claim for
full control of the louse Damalinia bovis, and it shares with the injectable
formulation a claim against adult Trichuris sp. More remarkable is its
approval for control of the adult (non-parasitic) horn fly, Haematobia
irritans, for 35 days after treatment.
(b) Sheep
1. IVOMEC Injection: a sterile solution as for cattle, for use at 200 pg
ivermectin kg-’ (not approved for use in the USA).
2. IVOMEC or ORAMEC Oral Liquid: a solution containing 0.08%
ivermectin, to be administered so as to give a dosage of 200 pg
ivermectin kg- ’.
Treatment with these products provides efficacy against intestinal and
pulmonary nematodes, as shown in Table 2. The injectable formulation
also gives efficacy against the mites Sarcoptes scabiei, Psorergates ovis
and Psoroptes ovis. The oral formulation, although active against all stages
of the nasal bot, Oestrus ovis, is not effective against mites other than
Psoroptes ovis.
(c) Swine
1. IVOMEC Injection: a sterile solution containing 1% ivermectin, as for
cattle and sheep, but intended to be administered in amount sufficient to
give a dosage of 300 pg ivermectin kg-’. For young pigs a dilute
solution (0.27%)permits greater dosing accuracy.
2. IVOMEC Premix for Swine: a premix containing 0.6% ivermectin. It is
Table 2 Parasites of sheep for which the use of ivermectin has been approved.
Gastrointestinal Other Nasal Itch
nematodes nematodes bOtS mites
Haemonchus contortusabC Gaigeria pachyscelisab Dictyocaulus filariaab Oestrus ovisab Psorergates ovisab
(adult & immature) (adult & immature) (adult & immature) (all larval stages)
Haemonchus placei" Oesophagostomum columbianumab Psoroptes ovisab
(adult) (adult & immature)
Ostertagia circumcinctaabC Oesophagostomum venulosumab
(adult & immature) (adult)
Trichostrongylusaxei Nematodirus baftus"
(adultb& immatureab) (adult & immature)
Trichostrongyluscolubriformisab Nematodirus spathiger
(adult & immature? (adult" & immatureab)
Cooperia curticei" Strongyloides papillosus
(adult & immature) (adult" & immatureab)
Cooperia oncophora" Chabertia ovina
(adult) (adult & immature)
Trichuris ovisab
(adult)
" Oral formulation.

Injectable formulation.
Including hypobiotic larvae.
Table 3 Parasites of swine for which the use of ivermectin’ has been approved.
Gastrointestinal nematodes Other nematodes Lice Mites
Ascaris suum (adult & 4 ) Metastrongylus spp. Haemotopinus Sarcoptes
(adult) suis scabiei
Hyostrongylus rubidus
(adult & L4)
Oesophagostomum spp.
(adult & 4 )
x
Strong loides ransomi
(adult)
’ Injectable formulation.
Colostral transmission to piglets can be prevented by treatment of sow 7-14 days
before farrowing.
intended for incorporation into swine feed, so as to give a dosage of

100 pg ivermectin kg-’ day-’.
These formulations are effective against intestinal, pulmonary, and renal
worms, as indicated in Table 3. Ivermectin is also active against the louse
Haematopinus suis and the mite S . scabiei. A regular control program is
needed to achieve ectoparasite control on a herd basis.
(d) Horses
1. EQVALAN Liquid: a solution containing 1% ivermectin and intended
for use as an oral drench or by nasogastric intubation. The recom-
mended dosage is 200 pg ivermectin kg-I.
2. EQVALAN Paste and ZIMECTERIN Paste: a paste formulation con-
taining 1.87% ivermectin and sold in a pre-filled syringe for oral
delivery of ivermectin at 200 pg kg-I.
The use of these formulations gives efficacy against the parasites listed in
Table 4. The list includes the migratory larvae of the nematode Strongylus
vulgaris and the skin-dwelling larvae of Onchocerca cervicalis. Treatment
is highly effective against stomach bots, the parasitic larvae of the flies
Gastrophilus intestinalis and Gastrophilus nasalis.
(e) Dogs
1. HEARTGARD-30 and HEARTGARD-30 Plus: tablets containing iver-
mectin in amounts of 65-272 pg, to be dispensed by veterinarians-for
administration by clients to their dogs on a monthly basis. The “plus”
designation indicates the addition of pyrantel pamoate. Administration
of tablets appropriate in size to that of the dog being treated will provide
an ivermectin dose of 6 pg kg-I. Dogs given the “plus” tablets will
also receive pyrantel pamoate at a dosage of 5 mg kg-I.
Table 4 Parasites of horses for which the use of ivermectina has been approved.
Gastrointestinal nematodes Other nematodes Bots
Strongylus vulgaris (adult & immature) Dictyocaulus arnfreldi Gastrophilus spp.
(adult & immature) (oral & gastric
stages)
Strongylus edentatus (adult & immature) Onchocerca sp.
(microfilaria)
Strongylus equinus (adult)
Triodontophorus spp. (adult)
Cyuthostomum spp. (adult & immature)
Cylicocyclus spp. (adult & immature)
Cylicostephanus spp. (adult & immature)
Cylicodontophorus spp.
(adult & immature)
Gyafocephalus sp. (adult & immature)
Oxyuris equi (adult & immature)
Parascaris equorum (adult)
Trichostrongylus axei (adult)
Habronema muscae (adult)
Strongyloides westeri (adult)
a Oral formulation.
2. HEARTGARD-30 Chewable: disks of soft moist material containing

ivermectin, as above, but formulated so as to make them palatable to
dogs. They are eaten when proffered instead of being swallowed when
inserted into the mouth.
Administration of ivermectin on a monthly basis will prevent heartworm
disease. This results from destruction of larvae of Dirofilaria immitis when
they are in the extravascular tissues of the dog. Both the third-stage and
fourth-stage larvae appear to be susceptible to the drug, but the subadult
and adult worms in the vascular system are not. Monthly treatment is
effective, not because of persistence of the drug in the tissues, but because
each treatment kills the larvae that have been introduced by mosquitoes at
riny time during the preceding month.
The dosage used for the prevention of heartworm disease is not sufficient
for the control of the common intestinal worms of dogs. However, the
“plus” formulation, which includes pyrantel pamoate, is effective against
intestinal ascarids (Toxocara canis and Toxascaris leonina) and hookworms
(Ancylostoma caninum and Uncinaria stenocephala).
4.3. Abamectin
Abamectin or avermectin B, is the naturally occurring precursor of

ivermectin. It differs from the latter only in having a double-bond at
the C22-23 position, whereas that linkage has been reduced by selective
hydrogenation in the case of ivermectin. The only approved commercial
formulation of abamectin is AVOMEC Injection for Cattle (Merck & Co.,
Inc.). It is similar to the corresponding ivermectin product and is not
further discussed here.
4.4. Moxidectin
Moxidectin is a derivative of nemadectin, a naturally occurring product of

the filamentous bacterium Streptomyces cyaneogriseus noncyanogenus.
These compounds are structurally similar to the milbemycins, and both
classes differ from the avermectins in the absence of a side-chain at the
C- 13 position. Moxidectin and nemadectin differ from the milbemycins
in havidg an unsaturated (2-25 chain (Carter, 1989). The structure of
moxidectin is shown in Figure 2.
4.4.1. General Properties

Nemadectin is a white fluffy solid when obtained by freeze-drying from
t-butanol. It is soluble in organic solvents and nearly insoluble in water
(Carter, 1989).
Moxidectin, the derivative in commercial use, has a broad spectrum of
antiparasitic activity and a wide margin of safety. According to a technical
brochure issued by the manufacturer (American Cyanamid Co.), no deaths
occurred in cattle given moxidectin at dosages up to ten times the recom-
mended dosage. A dosage of twice the recommended dosage was given to
bulls and pregnant cows but did not impair reproductive performance. In
safety trials in sheep, dosages up to five times the recommended figure did
not cause adverse reactions. At the recommended dosage, the drug has been
used safely in rams and pregnant ewes. In the course of efficacy trials in
horses, adverse reactions were not seen in horses given moxidectin at
dosages up to 400 pg kg-'.
According to the same brochure, nemadectin is much less toxic than
abamectin against the dung beetle Onthophagus gazella. Thus the concerns
raised about the possible adverse environmental impact of the avermectins
might be less applicable, or non-applicable, in the case of moxidectin.
4.4.2. Antiparasitic Utility

(a) Cattle. A number of commercial preparations are either marketed or
under development. The trademark cited has been registered by American
Cyanamid Co.
Table 5 Parasites of cattle for which the use of moxidectin" has received governmental approval. Where treatment is cited as
"aids in the control of" that too reflects regulatory approval.
Gastrointestinal worms Pulmonary Ticks Lice Mites Cattle
(adult and immature stages) worms grubs
Haemonchus placei Dictyocaulus Boophilus Linognathus vituli Psoroptes communis Hypoderma bovis
viviparus microplus var. bovis
Haemonchus contortus Haematopinus Sarcoptes bovis Hypoderma
eurysternus lineatum
Ostertagia ostertagi Solenopotes
(including inhibited larvae) capillatus
Ostertagia lyrata Damalinia bovis
(aids in control)
Ostertagia leptospicularis
Trichostrongylus axei
Trichostrongylus colubriformis
Trichostrongylus longispicularis
Cooperia oncophora
Cooperia pectinata
Cooperia punctata
Cooperia mcmasteri
Cooperia spatulata
Nematodirus helvetianus
Nematodirus spathiger
Oesophagostomum radiatum
Oesophagostomum venulosum
Bunostomum phlebotomum
Trichuris ovis
Trichuris discolor
Strongyloides papillosus
Toxocara vitulorum
Paramphistomum spp.
a Injectable formulation.
1. CYDECTIN Injection: a sterile solution containing 1% moxidectin. It is

injected subcutaneously so as to give a dosage of 200 pg moxidectin
kg-’.
2. Other products: products currently under development include a
“pour-on” solution and a sustained-release ruminal bolus.
Administration of the injectable product gives high efficacy against
many nematode and arthropod parasites, and these are listed in Table 5.
It can be seen that the drug is active against gastrointestinal and pulmonary
nematodes, against immature as well as mature worms, and against
inhibited fourth-stage larvae. The susceptible arthropods include ticks,
lice, mites and grubs. As expected with blood-borne drugs, moxidectin is
substantially more effective against sucking lice than against biting lice.
Data in support of commercial registration are on file in regulatory
archives. Review articles such as those available for ivermectin have not
yet started to appear for moxidectin, so the following references are cited
as sources of specific information: broad-spectrum activity against gastro-
intestinal nematodes: Ranjan et al. (1992), Samson et al. (1992), Williams
et al. (1992a); lungworm: Eysker and Boersema (1992), Ranjan et al.
(1992), Williams et al. (1992b); grubs: Scholl et al. (1992); lice: Webb
et al. (1991).
Ectoparasite efficacy data for an investigational slow-release bolus have
been published (Webb et al., 1991). Ruminal boluses containing 375, 750
or 1125 mg moxidectin were administered to cattle by balling-gun (the
rate of release, nominal or actual, was not given). The boluses were of
identical gross weight. All three were effective against the sucking louse
Solenopotes capillatus, with efficacy being apparent at 4 weeks after
treatment and complete at 6 weeks. None of the boluses was effective
against the biting louse Bovicola bovis. Feces collected from the treated
(and non-treated) animals were tested for their ability to support the
development of the larval stages of the face-fly Musca autumnalis. Treat-
ment with the bolus containing the greatest amount of moxidectin
(1 125 mg) resulted in feces with sufficient excreted drug to suppress M .
autumnalis development by more than 90% during the first 2 weeks after
treatment. Feces collected at longer intervals, or feces from animals given
boluses with less drug, were progressively less effective.
The efficacy of an investigational “pour-on” formulation of moxidectin
has also been tested against ectoparasites on cattle (Lonneux and Losson,
1992). The formulation contained 0.5% moxidectin and was applied to the
hide, along the back of the animals, in an amount equivalent to 500 pg
kg-’. The animals had moderate to severe mange, caused by the mite P.
ovis. Treatment with the pour-on preparation (and with the standard
injectable preparation) resulted in 100% elimination of the mites and

resolution of clinical disease.
The efficacy of an experimental oral formulation has also been reported
(Zimmerman et al., 1992).
(b) Sheep. The commercial formulation now available is CYDECTIN
Oral Drench (American Cyanamid Co.), a formulation containing 0.1%
moxidectin. It is to be administered oraIly in amount sufficient to give a
dosage of 200 pg moxidectin kg-'. Treatment of sheep with this prepara-
tion provides efficacy against the broad spectrum of parasites listed in
Table 6. The list includes gastrointestinal and pulmonary nematodes and
the itch mite Psorergates ovis.
(c) Swine. Moxidectin products for swine are not available for commer-
cial use at this writing. Natural infections of S. scabiei have been controlled
by treatment of pigs with a pour-on (5%) solution of moxidectin, applied so
as to give dosages of 500 or 750 pg kg-' (Gundlach et al., 1992). Similar
efficacy was obtained with an injectable (1%) solution at a dosage of
400 pg kg-'. Examination of feces for helminth eggs indicated that
such treatments may also be effective in eliminating Ascaris suum and
Oesophagostomum dentatum.
(d) Horses. Moxidectin is not currently registered for commercial use in
horses, but an efficacy trial has been reported (Lyons et al., 1992b). The
Table 6 Parasites of sheep for which the use of moxidectin"has been approved.
Gastrointestinal parasites Pulmonary worms External parasites
Haemonchus contortus Dictyocaulus jilaria Psorergates ovis
Ostertagia circumcincta
(including inhibited larvae)
Ostertagia trifurcata
Trichostrongylusaxei
Trichostrongyluscolubriformis
Trichostrongylusvitrinus
Nematodirus battus
Nematodirus spathiger
Nematodirus filicollis
Cooperia curticei
Cooperia oncophora
Cooperia pectinata
Cooperia punctata
Oesophagostomum columbianum
Oesophagostomum venulosum
Chabertia ovina
Trichuris ovis
Strongyloides papillosus
" oral formulation.

findings of the study were considered preliminary because various batches,

formulations, and routes of administration were used in small numbers of
animals, the horses were not of uniform type, and two methods of parasito-
logical assessment were used. Nevertheless, it was clear that moxidectin at
200400 pg kg-' is active against the nematodes Parascuris equorum,
S. vulgaris, Strongylus edentatus, and Habronema muscae and against the
bot G. nasalis. There were lapses or inconsistencies in efficacy against
Oxyuris equi, the migrating larvae of S. vulgaris and the bot G. intestinalis.
4.5. Milbemycin B-41D
The milbemycins are produced by Streptomyces hygroscopicus aureola-

crimosus and are structurally equivalent to 13-deoxy-ivermectinaglycones.
Milbemycin B-41D (Figure 2) has been used commercially for control of
D. immitis, but it appears to have little if any current use. Its efficacy
against D . immitis was described by Tagawa et al. (1985).
4.6. Milbemycin Oxime
Dogs. Milbemycin oxime (Figure 2) is an 80:20 mixture of the & and

A3 oximes of milbemycin D. Its development arose from the discovery that
milbemycin D has prophylactic activity against D . immitis when given
orally at lo00 pg kg-', one or two months after inoculation (Tagawa et
al., 1985). The milbemycins, unlike the avermectins, are more potent
against intestinal nematodes than against heartworm. Commercial use of
milbemycin D was followed by the more widespread marketing of the
oxime, which was introduced by Ciba-Geigy Animal Health under the
tradename INTERCEPTOR.
When given monthly to dogs, milbemycin oxime was shown to be fully
effective against precardiac stages of D . immitis (Bater, 1989; Blagburn et
al., 1989; Grieve et al., 1989) and against adult A. caninum in dogs
(Bowman et al., 1991a). The dosage required for complete efficacy against
both parasites is approximately 500 pg kg-', given orally, once a month.
When given for three months, this dosage had little if any efficacy against
U. stenocephala, even though it was 100% effective against the other
hookworm (Bowman et al., 1991a). A single dose of up to 1.2 mg kg-'
also was shown to be ineffective against adult and immature U.steno-
cephala (Shoop et al., 1993a). Milbemycin oxime has been developed for
the strategic control of both D. immitis and A. caninum, and the efficacy
and safety data have been reviewed (Stansfield and Hepler, 1991). As in the
case of ivermectin, idiosyncratic toxicity in collies is not observed at the
dosage used commercially. The following product is approved for

commercial use in dogs.
INTERCEPTOR: color-coded tablets containing milbemycin oxime in
amounts of 2.3-23 mg. A dog given a tablet appropriate to its body
weight will receive a dosage of 500 pg milbemycin kg-I. The tablets are
to be administered orally, once a month, for control of D . immitis and adult
A . caninum.
4.7. Doramectin
*
Doramectin is a new macrocyclic lactone (Figure 2). It was discovered
as a result of studies in which a mutant strain of Streptomyces avermi-
tilis was used to produce avermectins with C-25 substituents different
from those found in avermectins produced by conventional strains of the
bacterium. After screening for antinematodal activity in vitro (Dutton et
al., 1991), selected compounds were evaluated in laboratory animals and
in cattle (Goudie et al., 1993). The compound chosen for development
was 25-cyclohexyl-5-0-demethyl-25-de( 1 -methylpropyl) avermectin A],,
or doramectin. It has been given the trade name DECTOMAX, and
commercialization by Pfizer Inc. is imminent.
Studies on formulation and pharmacokinetics of doramectin led to the
development of formulations for parenteral use in cattle (Wicks et al.,
1993). As with other members of this class, the plasma profile can be
modified by formulating the drug in vehicles that yield different rates of
absorption from subcutaneous tissue. Sesame oil was found to be a good
vehicle, especially when ethyl oleate was added (10%) to enhance persis-
tence of efficacy. Such oil-based formulations were well tolerated by cattle
and gave clinically significant plasma levels for 12 days or more after
injection.
Cattle. Data collected in 28 trials conducted in North America and
Europe showed that doramectin, given subcutaneously at 200 pg kg-I,
was highly effective against a broad spectrum of gastrointestinal nema-
todes and lungwoms (Jones et al., 1993). Both natural and experimental
infections were used in this series of trials. Doramectin was >99%
effective against Ostertagia ostertagi (including inhibited larvae), Oster-
tagia lyrata, Haemonchus placei, Trichostrongylus axei, T. colubriformis,
Cooperia oncophora (including inhibited larvae), Cooperia pectinata,
Cooperia punctata, Cooperia spatula ta, Cooperia surnabada, Bunostomum
phlebotomum, Strongyloides papillosus, Oesophagostomum radiatum, and
Dictyocaulus viviparus. The drug was 93-97% effective against Tricho-
strongylus longispicularis, Nematodirus spathiger and Trichuris spp. It was
approximately 75% effective against adult and immature Nematodirus
helvetianus. Trials against infections in cattle in Latin America confirmed

the broad-spectrum efficacy of doramectin when given subcutaneously at
200 pg kg-' (Eddi et al., 1993). In addition to the species previously
mentioned, Haemonchus similis, H . contortus and Trichuris discolor
were shown to be susceptible to treatment. Efficacy against adult N .
helvetianus was 98%. The authors point out that this is comparable to
the efficacy of ivermectin against this species (N. helvetianus being the
dose-limiting species for both drugs) and that it is greater efficacy than
expected on the basis of trials with experimentally induced infections.
Treatment was 100% effective against eyeworms, Thelazia spp. (Kennedy
aqd Phillips, 1993).
Doramectin, like other avermectins, has prophylactic as well as thera-
peutic activity when administered parenterally to cattle; that is, infective
nematode larvae may fail to establish in treated hosts, the degree of effect
depending on the parasite species, the elapsed time after treatment, and
unknown factors affecting efficacy in individual animals. In controlled
trials, worm counts showed that doramectin prevented establishment of
C. oncophora for 14-21 days (slightly less in some individuals); prevented
establishment of 0. ostertagi for 21-28 days; and establishment of D.
viviparus for at least 28 days. These intervals are longer than those
reported by other workers for ivermectin, and may permit strategic control
programs based on less-frequent treatment (Weatherley et al., 1993). The
prophylactic efficacy of doramectin against C. oncophora and 0. ostertagi
has also been demonstrated in trials in which efficacy was assessed by fecal
egg count (Vercruysse et al., 1993).
Doramectin, like other avermectins, is active against arthropod parasites.
In field trials against lice and mites, a single injection at a dosage of 200 pg
kg-' was found 100% effective against Psoroptes bovis, S. scabiei,
Haematopinus eurysternus, Linognathus vituli and S . capillatus and 82%
effective against Damalinia bovis (sharing with ivermectin a lesser efficacy
against biting lice than against sucking lice). The results of these trials were
reported by Logan et al. (1993).
The same standard dosage was reported 100% effective against first,
second and third instars of the warble fly Hypoderma bovis (Hendrickx et
al., 1993). Against the tropical warble fly, Dermatobia hominis, doramectin
was reported to be 100% effective in destroying established larvae in the
nodules of experimentally infected cattle, and treatment prevented the
establishment of new warbles for at least 35 days (Moya-Borja et al.,
1993a). The New World screwworm, Cochliomyia hominivorax, is also
susceptible to doramectin (Mo a Borja et al., 1993b). A single sub-
cutaneous dose, at 200 pg kg-r gave 100% protection against myiasis
in cattle in which first instar larvae had been placed in surgical incisions.
This remarkable efficacy was attributed to the intrinsic action of the drug
and the prolonged plasma profile obtained with the formulation used. In
comparison, a conventional dose of ivermectin protects against the Old
World screwworm, but not against the New (Benz et al., 1989).
Doramectin was highly active against the tick Boophilus microplus in
two laboratory studies using experimentally infected calves (Gonzales et
al., 1993). In one study, a single injection (200 pg kg-’) was given to calves
which had been exposed repeatedly to infection and which carried ticks of
various degrees of maturity. As expected, there was no “knock-down
effect”; that is, there was no elimination of engorged ticks, because such
ticks would not have ingested drug. However, as the other ticks ingested
blood over the ensuing days, they were affected by the blood-borne drug.
Only a few ticks succeeded in engorging in the days immediately following
treatment, and these ticks exhibited a progressive reduction in egg produc-
tion and egg hatchability. By eight days after treatment, the engorgement
and detachment of ticks had completely stopped (except for one infertile
tick that detached on day 11). In the other study, the single injection of
drug was given before the multiple exposures to B . microplus. The infec-
tive larvae were applied to calves at intervals over a period of 17 days after
treatment. Saline-treated control calves showed that the ticks first reached
the engorged and detachment stage on day 23 after treatment (the first
detached ticks presumably being those applied immediately after treat-
ment). In the drug-treated calves, however, no detachments were recorded
until day 38. Even then, and in the three days remaining until the end of
trial, ticks detached in very small numbers and with greatly reduced
fecundity and egg hatchability. Thus the drug had persisted in host blood
for at least 17 days in amounts sufficient to interfere with the development
of B. microplus.
5. PROSPECTIVE DRUGS
5.1. Paraherquamide
Paraherquamide (Figure 3) is an oxindole alkaloid of fungal origin. It was

isolated around 1980 as a metabolite of Penicillium paraherquei and
reported as a toxin (Yamazaki et al., 1981). About a decade later, a culture
of Penicillium charlesii was found to have anthelmintic activity in a routine
in vivo assay, and the active principle was found to be paraherquamide
(Ondeyka et al., 1990; Ostlind et al., 1990). Chemical derivitization studies
were immediately undertaken (Blizzard et al., 1989). An unidentified
Penicillium sp. was independently reported to produce anthelmintic
metabolites identified as paraherquamides (Blanchflower et al., 1991).
Figure 3 Structures of paraherquamide, PF1022A, dioxapyrrolomycin, and

clonostachydiol.
5.1.1. Efficacy in vitro and in Laboratory Animals

Paraherquamide was active against the free-living nematode C . elegans at a
concentration of 2.5 pg ml-', whereas its 24,25-dihydro derivative was
inactive even at 200 pg ml-' (Ondeyka et al., 1990). It was active against
third-stage H. contortus at 31 kg ml-' (Blanchflower et al., 1991).
Jirds (Meriones unguiculatus) were given paraherquamide 6 days after
inoculation with infective larvae of T. colubriformis. Single oral dosages as
low as 1.6 mg kg-' were at least 98% effective in removing the young
adult worms (Ostlind et al., 1990). This extraordinary potency is surpassed
only by the macrocyclic lactone anthelmintics.
5.1.2. Efficacy in Sheep

Paraherquamide was tested against experimental infections of several
nematode species in sheep (Shoop et al., 1990). Treatment was given as
a single oral dose and necropsy was performed 7 days after treatment.
Dosages of 0.5 mg kg-' or higher were at least 98% effective against adult
H. contortus, Ostertagia circumcincta", T. axei, T. colubriformis and
Cooperia curticei, and against inhibited fourth-stage larvae of Cooperia
sp. Such dosages were of uncertain efficacy against inhibited fourth-stage
larvae of 0. circumcincCa and were not significantly active against adult
Oesophagostomum columbianum. It was estimated that the EDg5 for 0.
columbianum would be greater than 4 mg kg-'. In these trials, the strain of
H. contortus was resistant to ivermectin and the T. colubriformis was
resistant both to ivermectin and benzimidazoles.
5.1.3. Efficacy in Cattle

In cattle, paraherquamide was tested against both gastrointestinal and
pulmonary nematodes (Shoop et al., 1992b). Calves were infected experi-
mentally and were necropsied 14 days after treatment with a single oral
dose of the drug. A dosage of 1 mg kg-' was at least 95% effective
against H. placei, 0. ostertagi, T. axei, T. colubriformis, C . oncophora,
N . helvetianus, 0 . radiatum and D . viviparus. However, it was active
against C . punctata only at 4 mg kg-I. Paraherquamide thus has been
shown to be active against roundworms in the abomasum, small intestine,
large intestine and lungs.
5.1.4. Efficacy in Dogs

In experimentally infected dogs (Shoop et al., 1991), paraherquamide at
dosages up to 2 mg kg-' showed little or no activity against A . caninum,
(I.stenocephala, T. leonina or Trichuris vulpis. It was effective against
',
Strongyloides stercoralis at 2 mg kg- but treatment was not tolerated.
5.1.5. Safety and Other Aspects

Studies of acute toxicity of paraherquamide in CD-1 mice (Shoop et al.,
1992a) indicated an LD50of approximately 15 mg kg-'. The drug was well
tolerated in shee at dosages of up to 10 mg kg-' and in calves at dosages
up to 4 mg kg-'(highest dosage tested). In dogs, however, toxicity was
evident at dosages as low as 0.5 mg kg-I, and a dosage of 10 mg kg-' was
rapidly lethal (Shoop et al., 1992a).
*As discussed by Lichtenfels and Hoberg (1993) and Lichtenfels et al. (1988), some
recent classifications of the Trichostrongylidae have recommended the use of Teladorsagia
rather than Ostertagia for the species, 0. circumcincta, Ostertagia trifurcata, and Ostertagia
davtiani. However, these authors note that further work is needed to clarify the generic-level
systematics of the Ostertagiinae. Therefore, we have elected to use Ostertagia for these
species, as the majority of existing work uses this designation.
The above data suggest paraherquamide could be used successfully

against H. contortus, 0. circumcincta and T. colubriformis and would
enjoy a therapeutic index of 33. The importance of this conclusion lies:
(a) in the recognized potential of those species to develop anthelmintic
resistance, and (b) in the likelihood that paraherquamide has a novel mode
of action and would therefore probably not be cross-resistant with existing
anthelmintics. The mode of action has not been determined, but novelty is
suggested by efficacy of the drug against strains resistant to ivermectin and
the benzimidazoles (Shoop et al., 1990).
5.2. PF1022A
PF1022A (Figure 3) is a cyclodepsipeptide of fungal origin. It was isolated

from a mycelial cake of Mycelia Sterilia PF1022. PF1022A is a colorless,
crystalline solid which is soluble in common organic solvents and insoluble
in water (Sasaki et al., 1992).
The anthelmintic activity of PF1022A was discovered in a screen using
chickens infected with Ascaridia galli where the compound cleared > 91%
of the worms at a dose of 2 mg kg-' (Sasaki et al., 1992). Information
available, to date, on efficacy against nematodes of veterinary importance
was summarized by Terada (1992) and Terada et al. (1993). It was noted
that in addition to activity against A . galli the compound is effective
orally against T. canis and Toxocara cati in dogs at 0.2 mg kg-',
effective against H . contortus orally (in cattle, dose not specified) or
intravenously (dose and host not specified), and effective against 0.
ostertagi in cattle (route and dose not specified), whereas in vitro the
motility of Heterakis spumosa is inhibited completely within 2 h of treat-
ment at g ml-'. Sasaki et al. (1992) indicated that PF1022A produced
no signs of acute toxicity in mice when administered at 1 g kg-' intra-
peritoneally or at 2 g kg-' orally.
5.3. Dioxapyrrolomycin
Dioxapyrrolomycin (Figure 3) was originally isolated as a metabolite from

a Streptomyces sp. The compound, obtained as fine yellow needles, is
soluble in common organic solvents and practically insoluble in water
(Carter et al., 1987).
Dioxapyrrolomycin was found to have anthelmintic activity as described
by Conder et al. (1992). Initially, a crude fermentation preparation of an
actinomycete culture (designated UC 11065) was found in routine assays to
have activity in vitro against C . elegans and subsequently in vivo against H .
contortus in the jird. The active component of the crude preparation was
identified as dioxapyrrolomycin, which was shown to clear >90% of H.
contortus from jirds when administered orally at 0.33 mg per animal, while
having only slight activity against T. colubriformis in the model (-40%
clearance at 0.33-1.0 mg per jird). In sheep, dioxapyrrolomycin given
orally at doses 23.125 mg kg-' cleared > 99% of H. contortus, whereas
>92% clearance was observed at 1.56 mg kg-'. Studies done in the jird
(Conder et ul., 1992) using ivermectinbenzimidazole-or levamisole/ben-
zimidazole-resistant strains of H. contortus demonstrated that dioxapyrro-
lomycin has approximately equal efficacy against the resistant and
susceptible strains, and so is not cross-resistant with the three major
classes of broad-spectrum anthelmintics (benzimidazoles, levamisole/
morantel/pyrantel, and macrocyclic lactones). However, an in vitro
migration assay showed dioxapyrrolomycin is -6 times less active
against closantel-resistant H. contortus than ,against susceptible worms.
Based on these studies dioxapyrrolomycin appears to be a narrow-spectrum
anthelmintic with a closantel-like mode of action. The LD50of dioxa-
pyrrolomycin in mice is 13 mg kg-I (Carter et al., 1987). No toxic signs
were observed at doses up to 12.5 mg kg-' (highest dose examined) in
sheep in the studies reported by Conder et al. (1992).
5.4. Clonostachydiol
Clonostachydiol (Figure 3) was ,identified as a metabolite of the fungus,

Clonostuchys cylindrospora. It is a colorless, crystalline solid which is
soluble in common organic solvents and insoluble in water or n-hexane
(Grabley et al., 1993). A dose of 2.5 mg kg-' given subcutaneously to
sheep artificially infected with H. contortus resulted on day 14 post-
treatment in an 80-90% reduction from pre-treatment egg counts (Grabley
et al., 1993).
6. RESISTANCE TO ANTINEMATODAL DRUGS
6.1. Is Resistance an Issue?
Drug resistance is here defined as: a heritable reduction in the sensitivity of

a parasite population to the action of a drug, the reduction being expressed
as a decrease in the frequency of individual parasites affected by exposure
to the drug (in comparison to the frequency observed in the same popula-
tion upon initial or prior exposure). Resistance should not be confused with
tolerance. In the context of veterinary parasitology, the term tolerance is
conventionally restricted to the innate unresponsiveness of a parasite
population to a drug; that is, to the insusceptibility of a population,
independent of prior exposure to that drug or others of its class. These
usages are consonant with those of Coles (1986b), Prichard et al. (1980)
and Shoop (1993). In general, discussion will be limited to the three classes
of modem broad-spectrum anthelmintics, i.e. benzimidazoles, levamisole/
morantel/pyrantel, and macrocyclic lactones.
Experience in the antibacterial/antiviral (Cohen, 1992; Gibbons, 1992,
Neu, 1992), insecticidal (Georghiou, 1986; Roush, 1990), and protozoal
(Chapman, 1990; Femex et al., 1990) arenas has clearly demonstrated
resistance is the rule rather than the exception. Therefore, it should come
as no surprise that resistance of nematodes to anthelmintics is a growing
problem, particularly in certain geographic areas and some host species.
Resistance may be expected to develop more slowly in nematodes than in
bacteria, viruses, insects, or protozoa. This is due to longer generation times,
a more limited range of mobility (essentially that of their host), selection
generally limited to parasitic stages, and the high efficacy and non-persistent
nature of most anthelmintics (in the case of persistence, the macrocyclic
lactones and the salicylanilides are exceptions). Nevertheless, anthelmintic
resistance is and will continue to be a problem. Waller (1990) points out
“. . . most of the important nematode genera of domestic livestock have
shown that they possess the genetic capacity to develop resistance”.
To exemplify that resistance to antinematodal drugs is indeed something
with which we should be concerned, the following six points should be
considered.
1. Resistance to benzimidazoles is present worldwide for small strongyles

in horses (Lyons et al., 1990), and there is some evidence that resistance
to pyrantel is developing to these parasites in the horse (Herd, 1992). It
is worth noting that small strongyles can induce clinical disease (Baker
et al., 1984; Giles et al., 1985; Herd, 1986a, 1992; Mair and Cripps,
1991).
2. Resistance to all classes of anthelmintics is extensively documented in
sheep and goats, and in some cases serious problems exist. For instance,
it has been reported (Van Wyk et al., 1989a, b) that some farmers in
South Africa are getting out of business simply because there are no
longer any drugs which are effective in controlling the multi-resistant
nematode populations. Rolfe (1990) reports, “Resistance to antiparasitic
agents in sheep has emerged as the most important limitation for
successful production of wool and sheep meat in Australia”, and
Waller et af. (1990b) suggest that “. . . resistance is widespread and
at a high level in the high rainfall regions of the East African (Kenya,
Tanzania, Zimbabwe) and South American (Argentina, Brazil, Uruguay)
countries”.
3. The problem of resistance is growing in both the number of properties
having resistance and the level of resistance observed. This is supported
by surveys conducted in earlier years compared to similar surveys done
more recently in New Zealand (West et al., 1989; McKenna et al.,
1990), South Africa (Van Wyk et al., 1987, 1990), England (Taylor,
1990a; Coles et al., 1991), Australia (Edwards et al., 1986; Waller e f al.,
1988; Love et al., 1992), and the Netherlands (Borgsteede, 1990;
Borgsteede et al., 1991).
4. Whereas in earlier surveys and clinical reports resistance was limited to
a single parasite and anthelmintic class, there are now instances of
multigeneric resistance (examples include Barton et al., 1985; Martin
et al, 1985; Edwards et al., 1986; Anderson et al., 1988a; Hughes, 1988;
Waller et al., 1988, 1990a; McKenna, 1989; West et al., 1989; Watson
and Hosking, 1990; Duwel, 1991; Hong et al., 1992; Varady et al.,
1993) and/or multidrug (class) resistance (examples include Sangster et
al., 1979; Green et al., 1981; Hall et al., 1981a; Kettle.et al., 1983;
Barton et al., 1985; Dash, 1986a; Edwards et al., 1986; Anderson et al.,
1988a, b; Drudge et al., 1988; Van Wyk and Malan, 1988; Waller et al.,
1988, 1990a; Van Wyk et al., 1989a, b; Badger and McKenna, 1990;
McKenna et al., 1990; Watson and Hosking, 1990; Maingi, 1991a;
Reinecke et al., 1991; Jackson et al., 1992a, b; Love et al., 1992;
Pomroy et al., 1992; Uppal et al., 1992; Varady et al., 1993).
5. Resistance is becoming evident in hosts, such as swine (Roepstorff et al.,
1987; Bj@m et al., 1989, 1990; Waller et al., 1990b) and cattle
(Anderson, 1977; Anderson and Lord, 1979; Boersema, 1983a; Eagleson
and Bowie, 1986; Geerts et al., 1987; Jackson et al., 1987; Pinheiro
and Echevarria, 1990; Hosking and Watson, 1991; McKenna, 1991;
Williams, 1991; Williams et al., 1991; Eagleson et al., 1992; Watson,
1993), where it was not previously recognized.
6. Unfortunately, resistance is generally not recognized until it becomes a
problem, due to our inability easily to assess subclinical resistance in the
field and to our general unwillingness to accept anything less than
clinical failure as an indication of resistance. Once resistance results
in clinical failure it is probably too late to do anything to reverse this
situation. Although there are a few examples of resistant strains
reverting to susceptibility following discontinued use of anthelmintics
of the class that elicited resistance, the overwhelming evidence suggests
that reversion is unlikely (for example Herlich et al., 1981; Hall et al.,
1982; Le Jambre et al., 1982; Herd et al., 1984; Dash, 1986b;
Boorgsteede and Duyn, 1989; Taylor and Hunt, 1989; Lyons et al.,
1990; Jackson, 1993), and even when it occurs, the increased suscept-
ibility is not sufficient to be of value in the field or there is rapid
reselection for resistance (Simpkin and Coles, 1978; Kelly and Hall,
1979; Martin el al., 1988a; Waller et al., 1988).
An interesting observation was made by Ian Barger (CSIRO Laboratory,
Armidale, NSW, Australia) at a meeting of European (France, Ireland,
Spain and UK) scientists where they were examining with colleagues
from Australia what had been learned from the Australian experience
with resistance which could be used to prevent similar problems in
Europe. His suggestion was that the problem had probably already deve-
loped too far in Europe to learn from the mistakes of the Australian sheep
industry (Anonymous, 1992). The same may be said for much of the rest
of the world. Clearly, based on the above, resistance is an issue. It will
not go away and it will continue to worsen without significant and timely
intervention.
6.2. Extent of Resistance Worldwide
Although an attempt has been made to detail by country and host those
species of nematodes which have been reported to be resistant to one or
more of the three classes of modem broad-spectrum anthelmintics, there
are undoubtedly reports which have been missed, particularly where the
literature relating this information is hard to access. Reports of resistance
where the resistant nematode species (or genera) are not identified will not
be detailed herein, except in the case of the horse where only rarely are
identifications made. In addition, it is often difficult to assess whether
resistance was indeed present based on the information provided, hence
our inclusion or omission of particular reports of resistance may not agree
entirely with others’ interpretations of the data.
6.2.1. Sheep and Goats

Historically, resistance has emerged rapidly in sheep and/or goats for each
new class of antinematodal drugs. Although many factors contribute to
nematode resistance in these host species, resistance is probably primarily
due to two important aspects of management, i.e. frequent dosing, parti-
cularly in warm, moist climates, and the practice of running sheep and
goats together. Both factors are discussed in section 6.5, but it is worth
noting here the obvious but often ignored fact that sheep and goats are
different animals. It is clear that anthelmintic metabolism is quite different
in these two species, and higher doses of anthelmintics appear to be
Table 7 Nematodes of sheep and/or goats with reported resistance to broad-spectrum anthelmintics.
Anthelmintic class'
Nematode country Benzimidazoles Levamisole/morantel/pyrantel Macrocyclic lactones
Haemonchus contortus Australia 1,2,3,4,5,6,7,8,9,10 5,820 11
Belgium 12.13 - -
Brazil 14,15,16,17 15,18,19 16,17,20
France 21,22,23 - -
Germany 24,25,26,27 - -
India 28,29,30 29,30,3 1 -
Kenya 32 33' -
Malaysia 34 - -
Martinique 35,36 - -
Netherlands 37,38,39,40 - -
New Zealand 4 1,42,43,44,45,46 47' -
South Africa 48,49,50,51,52,53 53 51,52,54,55
Sri Lanka 56 - -
Switzerland 57 - -
Tanzania 58,59,60 - -
United Kingdom 61,62,63,64,65,66,67,68,69 - -
United States 70,71,72,73,74,75,76,77,78,79, 86 82
80,81,82,83,84,85
Ostertagia circumcincta Australia 6,9,10,87,88,89,90,91 10,88,91,92 93'
Brazil - 19 -
Czechoslovakia 94'Vd 94c.d 94'"
Denmark 95' 95 -
France 21,22,23,96,97 - -
Germany 27 - -
Netherlands 3940' - -
New Zealand 43,45,98,99 98,99 98,99,100
South Africa 101 101 102'
United Kingdom 64,65,66,68,103,104,105,106, 109' 108
107,108
United States 79,80 -
Ostertagia trifurcata Australia 9,89,90 92

France 23 -
New Zealand 45,98,99 98,99
Ostertagia davtiani Australia 9 -
Trichostrongylus
colubriformis Australia 6,7,9,10,88,91,110,111 6,7,9,10,88,91,111,112
Brazil - 15,18,113
Czeckoslovakia 94'vd 94'Td
Denmark 95' 95'
France 21,22,23,114 -
Germany 27
Kenya 115
Netherlands 39,40' - -
New Zealand 43,45 116 99,117'
South Africa - 118 -
United Kingdom 64
United States 80
Trichostrongylus axei Australia 9
France 21 -
United Kingdom - 109
Trichostrongylus vitrinus Australia 9 9
New Zealand 45
Table 7 continued
Anthelmintic classa
Nematode Country Benzimidazoles L.evamisole/morantel/pyrantel Macrocyclic lactones
Nematodirus spathiger Australia 9,90,119,220
Brazil 121
France 21'
New Zealand 45,112
South Africa 101
Nematodirus Jilicolis Australia 9-90
New Zealand 45
Nematodirus abnormalis Australia 9,120
Cooperia curticei France 23,96,123
Netherlands 124
New Zealand 125'
United Kingdom 126,127
Oesophagostomum columbianum Australia 8'
Brazil -
United Kingdom 128'
Oesophagostomum venulosum France 21
New Zealand 45
Chabertia spp. New Zealand 125
United Kingdom 64
Strongyloides papillosus Brazil -
New Zealand 49
a References: (1) Smeal et al., 1968, (2) Le Jambre et al., 1979a, (3) Webb et af., 1979, (4), Barton, 1980, (5) Green et al., 1981, (6)
Barton et af.,1985, (7) Gillham and Obendorf, 1985, (8) Edwards et al., 1986, (9) Beveridge et al., 1990, (10) Love et al., 1992, (11) Le
Jambre, 1993a, (12) Vercruysse et al., 1989, (13) Geerts et al., 1990, (14) dos Santos and Franco, 1967, (15) Charles et al., 1989, (16)
Echevarria et af., 1991, (17) Vieira et al., 1992, (18) Santiago and da Costa, 1979, (19) Santiago et al., 1979, (20) Echevarria and
Trindade, 1989, (21) Kerboeuf et al., 1988, (22) Kerboeuf and Hubert, 1990, (23) Hubert et af., 1991, (24) Bauer et af., 1987, (25)
Duwel et af., 1987, (26) Bauer et al., 1988, (27) Diiwel, 1991, (28) Yadav, 1990, (29) Uppal et af.,1992, (30h Yadav et al., 1993, (31)
Yadav and Uppal, 1992, (32) Njanja et af.,1987, (33) Maingi, 1991a, (34) Domy et af., 1993, (35) Gruner et al., 1986, (36) Bastien et
al., 1989, (37) Boersema et al., 1982, (38) Eysker et al., 1983a, (39) Boersema et al., 1987, (40) Borgsteede et al., 1991, (41) Vlassoff
and Kettle, 1980, (42) Kemp and Smith, 1982, (43) Kettle et af.,1982, (44)McKenna and Hughes, 1986, (45) McKenna, 1989, (46)
Scherrer et al., 1989, (47) Kettle et af., 1983, (48) Berger, 1975, (49) Van Schalkwyk, 1984, (50) Van Wyk et a[., 1987, (51) Van Wyk
and Malan, 1988, (52) Van Wyk et al., 1989a, (53) Van Wyk et al., 1989b, (54) Anonymous, 1986, (55) Cannichael et af., 1987, (56)
Van Aken et af., 1989, (57) Jordi, 1980, (58) Kassuku and Tibaijuka, 1987, (59) Ngomuo et al., 1990, (60) Bjarn et af., 1991a, (61)
Cawthorne and Cheong, 1984, (62) Taylor and Hunt, 1988, (63) Scott et af.,1989, (64) Scott el al., 1990, (65) Coles et al., 1991, (66)
Jackson et al., 1991, (67) Fisher et al., 1992b, (68) Hong et al., 1992, (69) Jackson et al., 1992c, (70) Conway, 1964, (71) Drudge et al.,
1964, (72) Knight et al., 1967, (73) Shelton, 1968, (74) Colglazier et al., 1969, (75) Partosoedjono et al., 1969, (76) Colglazier et af.,
1970, (77) Theodorides et al., 1970, (78) Andersen and Christofferson, 1973, (79) Miller and Baker, 1980, (80) Herd et al., 1984, (81)
Uhlinger et al., 1988, (82) Craig and Miller, 1990, (83) Lyons et al., 1992a, (84) Uhlinger et af., 1992, (85), Lyons et af.,1993, (86)
Miller et af.,1987, (87) Hall et al., 1979, (88) Sangster et al., 1979, (89) Riffkin et af., 1984, (90) Martin el al., 1985, (91) Waller et af.,
1988, (92) Le Jambre, 1979, (93) Besier and Wroth, 1993, (94) Varady et al., 1993, (95) Bjem et al., 1991b, (96) Gevrey et al., 1984,
(97) Beugnet, 1992, (98) Watson and Hosking, 1990, (99) Pomroy et al., 1992, (100) Badger and McKenna, 1990, (101) Van
Schalkwyk and ShrWer, 1989, (102) Reinecke et al., 1991, (103) Britt, 1982, (104) Cawthome and Whitehead, 1983, (105) Mitchell et
al., 1991, (106) Orpin, 1991, (107) Scott etal., 1991a, (108) Jackson etal., 1992b, (109) Britt, 1986, (110) Hotson etal., 1970, (111)
Dash, 1986a, (112) Waller et al., 1986, (113) Santiago et al., 1977, (114) Cabaret, 1991, (115) Maingi, 1991b, (116) McKenna and
Seifert, 1985, (117)McKennaetaf., 1990, (118)Van Wyketal., 1990,(119) Obendorfet al., 1986, (120)Obendorfetal., 1991,(121)
da Costa et al., 1985, (122) Middelberg and McKenna, 1983, (123) Dorchies et al., 1991, (124) Borgsteede, 1986, (125) Hughes, 1988,
(126) Taylor, 1990a, (127) Hunt et al., 1992, (128) Scott and Mitchell, 1990, as reported by Scott el al., 1990.
-, NO known reports.
Genus but not species identified.
The animals were imported from New Zealand.
required but are generally not used in goats (McKenna and Watson, 1987;
Coles et al., 1989a; Scott et al., 1989; Sangster et al., 1991a; Hennessy et
al., 1993). In short, treating goats as though they were sheep and com-
mingling these species can contribute to anthelmintic resistance. Both hosts
are considered together here, because of the role this intimate relationship
between sheep and goats has had on resistance development.
Table 7 shows those parasite species by geographic distribution which
have been identified as being resistant to antinematodal drugs from the
three modem broad-spectrum classes in sheep and/or goats. Haemonchus
contortus, Ostertagia spp. and Trichostrongylus spp. are resistant to the
benzimidazoles throughout much of the world but show a more spotty
distribution of resistance to levamisole/morantel/pyrantel and the macro-
cyclic lactones. Resistant parasites in other genera (Nematodirus, Cooperia,
Oesophagostomum, Chabertia and Strongyloides) show more limited
distributions, which may reflect innate differences in ability to generate
viable resistant populations for these organisms or may be an artifact of
inadequate recognition due to the relatively lower pathogenicity and hence
notice of these species compared to more problematic species. It also is
evident that the narrow-spectrum salicylanilides, used almost exclusively
to control strains of H. contortus resistant to the broad-spectrum anthel-
mintics, are rapidly generating resistance in South Africa (Van Wyk and
Gerber, 1980; Van Wyk et al., 1982, 1987) and Australia (Rolfe et al.,
1990).
6.2.2. Cattle
In contrast to the situation in sheep and goats, resistance has been slow to
develop in cattle. Barger (1993) suggests that bovine dung-pats may
provide a relatively larger refugia of susceptible infective larvae, and
hence, reduce the proportion of the population exposed to anthelmintic
selection. Alternatively, less frequent use of anthelmintics in this host may
minimize selection pressure. It has been suggested (Eagleson and Bowie,
1986; Waller, 1993) that selection for resistance in some parasites of cattle
occurs in other species, such as goats, which can harbor species of
nematodes common to both hosts. As noted in Table 8, few reports of
resistance are available, but in many cases the parasite identified as being
resistant is 0. ostertagi. Since, with a single exception, all of the reports of
resistance to date are for levamisole/morantel or benzimidazoles, both of
which provide less than optimal protection against inhibited larval stages, it
is possible that resistance has developed in response to what amounts to
subtherapeutic dosing of inhibited larvae. Populations of Haemonchus spp.,
T. axei and C . oncophora resistant to the benzimidazoles have also been
identified sporadically, Boersema ( 1983a) reported a strain of Dictyocaulus
Table 8 Nematodes of cattle with reported resistance to broad-spectrum

anthelmintics.
Anthelmintic Class'
Nematode Country Benzimidazoles Levamisole/ Macrocyclic

moranteV lactones
pyrantel
Haemonchus spp. Brazil 1 -b -
Ostertagia
ostertagi Australia 23 2 -
Belgium - 4 -
New Zealand 5 - -
United States - 6.7 -
Trichostrongylus
axei Australia 8-9 - -
New Zealand 10' - -
Cooperia
oncophora New Zealand 5,ll - 12'
Dictyocaulus
viviparus Belgium - 13 -
' References: (1) Pinheiro and Echevarria, 1990, (2) Anderson, 1977, (3) Anderson
~ ~~
and Lord, 1979, (4) Geerts et al., 1987, ( 5 ) Hosking and Watson, 1991, (6) Williams,
1991, (7) Williams et al., 1991, (8) Eagleson and Bowie, 1986, (9) Eagleson et al.,
1992, (10) McKenna, 1991, (11) Jackson et al., 1987, (12) Watson, 1993, (13)
Boersema, 1983a.
-, NO known reports.
' Genus but not species identified.
viviparus resistant to levamisole, and Watson (1993) has identified a

Cooperia sp. resistant to ivermectin.
6.2.3. Horses
Small strongyles are recognized throughout the world as being resistant
to the benzimidazoles (Table 9). To date, 13 species (Cyathostornurn
catinatum, Cyathostornum coronaturn, Cyathostomum labiaturn, Cyathos-
tornurn labratum, Cylicocyclus brevicapsulatus, Cylicocyclus insigne,
Cylicocyclus leptostomus, Cylicocyclus nassatus, Cylicostephanus cali-
catus, Cylicostephanus goldi, Cylicostephanus longibursatus, Cylico-
stephanus minutus, and Cylicostephanus poculatus) have been identified
with benzimidazole resistance. In addition, resistance to pyrantel is
believed to be present for small strongyles in the United States (Herd,
1992). Benzimidazole resistance appears to be present in the United States
Table 9 Nematodes of horses with reported resistance to broad-spectrum anthelmintics.
Anthelmintic class'
Nematode country Benzimidazoles Levamisole/morantel/pyrantel Macrocyclic lactones
Cyathostomwn catinatum Germany 1
United States 2,3,4,5,6,7
Cyathostomum coronatum Belgium 8
Germany 1
Netherlands 9,lO
United States 2,3,4,5,6,7,11
Cyathostomwn labiatum Belgium 8
Netherlands 10
United States 2
Cyathostomum labratum Belgium 8
Netherlands 9,lO
Cylicocyclus brevicapsulatus United States 11
Cylicocyclus insigne United States 2,ll
Cylicocyclus leptostomus United States 5,ll
Cylicocyclus nassatus Belgium 8
Germany 1
Netherlands 9,lO
United States 2,3,4,5,6,7,11
Cylicostephanus calicatus Belgium 8
Germany 1
Netherlands 9,lO
United States 54
Cylicostephanus goldi Belgium 8
Germany 1
United States 2,4,5,6,7,11
Cylicostephanus longibursatus Belgium
Germany
Netherlands
United States
Cylicostephanus minutus Germany
Netherlands
United States
Cylicostephanus poculatus Germany 1
Unspecified small strongyles" Australia 12,13
Austria 14
Brazil 15,16,17
Canada 18,19
Denmark 20
New Zealand 21
Norway 22,23
South Africa 24,25
Sweden 26
United Kingdom 27,28,29,30,3 1,32,33,34
Strongylus edentatus United States 35
aReferences: (1) Burger and Bauer, 1987, (2) Drudge et al., 1977, (3) Drudge et al., 1983, (4) Drudge et al., 1984, ( 5 ) Wescott et al.,
1985, (6) Chapman et al., 1991, (7) Drudge et al., 1991, (8) Geerts et al., 1988, (9) Eysker et al., 1988, (10) Eysker et al., 1989, (11)
Wescott et al., 1982, (12) Barger and Lisle, 1979, (13) Kelly et al., 1981b, (14) Lippert and Prosl, 1988, as reported by Borgsteede,
1990, (15) Vieira-Bressan et al., 1988, (16) Campos Pereira et al., 1989, (17) Campos Pereira et al., 1991, (18) Slocombe and Cote,
1977, (19) Slocombe et al., 1989, (20) Bj0m et al., 1991c, (21) Hope and Kemp, 1980, (22) Helle, 1986, (23) Waller et al., 1990b, (24)
Van Wyk, 1987, (25) Van Wyk and Van Wijk, 1992, (26) Nilsson et al., 1989, (27) Round et al., 1974, (28) Herd, 1986c, (29) Ryan et
al., 1987, (30) Britt and Clarkson, 1988, (31) Love et al., 1989, (32) Lumsden et al., 1989, (33) King et al., 1990, (34) Fisher et al.,
1992a, (35) Uhlinger and Johnstone, 1984.
-, No known reports. ' Includes only those countries where the resistant species have not been identified in at least one report.
for S . edentatus (Uhlinger and Johnstone, 1984), although the authors

indicated further studies were needed to confirm this finding.
6.2.4. Swine
In Europe, A. suum resistant to benzimidazoles and Oesophagostomum
quadrispinulatum and 0. dentatum resistant to pyrantel citrate have been
reported (Table 10). It is interesting to note that in the case of Oesophago-
stomum spp. modem management practices have virtually eliminated the
parasite refugia, resulting in infection coming almost uniformly from
offspring of resistant worms surviving treatment (Roepstorff et al., 1987).
6.2.5. Other Hosts

Reports of resistance for nematodes in other hosts are rare. Examples
include Libyostrongylus douglassi resistant to levamisole in the ostrich
in South Africa (Malan et al., 1988) and H . contortus resistant to benzi-
midazoles in Roan antelope in the United States (Isaza et al., 1987). We are
not aware of any documented reports in people, dogs or cats.
6.3. Causes of Treatment Failure
If we are to have any chance to control or at least minimize resistance, we

must first be able to recognize the problem as early as possible and then
utilize and continue to improve on appropriate control strategies. Two
inherent problems are associated with the first critical step of recognizing
Table 10 Nematodes of swine with reported resistance to broad-spectrum

anthelmintics.
Anthelmintic class'
, Nematode Country Benzimidazoles Levamisole/ Macrocyclic

moranteV lactones
pyrantel
Ascaris mum Denmark 1 -b -
Oesophag ostomum
dentatum Denmark - 2,3,4 -
Oesophagostomum
quadrispinulatum Denmark - 2-34 -
' References: ( 1 ) Waller et al., 1990b, (2) Roepstorff et al., 1987, (3) Bjarn et al.,
1989, (4) Bjam et al., 1990.
-, No known reports.
CHEMOTHERAPY OF NEMATODE INFECTIONS OF VEJERINARY IMPORTANCE 39
resistance, i.e. eliminating those causes of treatment failure which are

frequently and incorrectly attributed to resistance and correctly identifying
resistance early on and prior to the appearance of clinical problems. The
former problem is at least theoretically controllable through appropriate
educational programs, whereby producers and clinicians/scientists work
together to better understand those factors unrelated to resistance which
can lead to treatment failure, The latter problem, i.e. timely recognition
of resistance, has proven to be more intractable, both technically and
philosophically.
There are a number of causes of treatment failure which are unrelated to
resistance. The following six examples are provided.
1. Misdiagnosis is clearly at the head of this list. A variety of other

infections (protozoal, bacterial, and viral), disease syndromes (mineral
deficiency or plant toxicosis), or even a change in diet can mimic the
clinical signs of nematode infections; obviously an antinematodal drug
will be ineffective in these situations.
2. Underdosing animals is a common cause of treatment failure. This
practice occurs frequently and, in some cases, intentionally. Where
intentional, producers generally believe the drug is effective at a lower
than recommended dose and, in an effort to cut costs, use the lower
dose. Unintentional underdosing usually occurs as a result of incorrectly
estimating animal weights. A study (Besier and Hopkins, 1988) in
Australia, where farmers were asked to estimate the weight of sheep,
indicated that only 27% of those surveyed were within 20% of the
correct weight and 86% of estimates were below the correct weight,
which would have resulted in underdosing by as much as 65% in some
animals. Not only does underdosing lead to treatment failure, it also sets
the stage for rapid development of resistance, since it allows not only
homozygous but heterozygous resistant individuals to survive treatment.
In some cases, animals also are inadvertently not dosed.
3. Since most antinematodal drugs do not have persistent activity,
macrocyclic lactones and salicylanilides being the obvious exceptions,
it is possible for treated animals to become rapidly reinfected. In this
case, successful treatment can be interpreted as treatment failure. This is
particularly a problem where animals are being treated for clinical
disease resulting from repeated and massive exposure as a result of
local epidemiological conditions and/or of poor management.
4. Physiological phenomena can contribute to treatment failure. For
example, the esophageal groove reflex can result in drug getting into
an inappropriate compartment where it is rendered ineffective or less
effective.
5. Pharmacokinetic differences between individual animals and between
species can have dramatic effects on treatment outcome. As noted above

goats are often treated as though they were sheep, although recent
studies with levamisole (McKenna and Watson, 1987; Coles et al.,
1989a) and benzimidazoles (Sangster et af., 1991a), have demonstrated
that higher doses are required for efficacy in goats than in sheep. The
higher dose requirement for efficacy in goats is not surprising given the
differences in drug pharmacodynamics (Gillham and Obendorf, 1985;
Bogan et al., 1987; Hennessy et al., 1993) between the two host species
and the degree of rumen bypass in goats (Sangster et al., 1991a). Scott
et al. (1990) also demonstrated that bioavailability of ivermectin may be
reduced in goats compared to sheep. Likewise, elevated doses are
required for efficacy of ivermectin in red deer where Andrews et al.
( 1993) demonstrated that the pharmacokinetics of ivermectin differed
dramatically from those of cattle, i.e. lower plasma peak concentrations
and shorter half-life in red deer. Similar results have been observed for
oxfendazole in red deer compared to sheep (Watson and Manley, 1985).
In addition, other factors such as disease or diet can play a significant
role in the pharmacokinetics of drugs. This latter point was demon-
strated by Taylor et al. (1992) who showed that fenbendazole and
ivermectin in pastured lambs and fenbendazole in pastured calves had
significantly lower peak plasma concentrations and areas under the
plasma concentration-time curve (i.e. reduced drug availability) than
did housed animals fed hay and concentrates.
Use of inappropriate drugs to deal with particular problems is yet
another cause of treatment failure. A good example of this is the use
of an antinematodal drug without a claim for inhibited larvae in situa-
tions or areas where these larvae are major contributors to disease. It
should be obvious that the potential for severe pathogenesis is substantial
when only adults and developing larvae are effectively controlled with
the drug being used, particularly given the typically large numbers of
inhibited larvae which can be present under hypobiotic conditions. An
appropriate drug which has been mishandled (outdated or improperly
stored, mixed or delivered) may also result in treatment failure.
This outline of causes for treatment failure unrelated to resistance might

suggest that the problem of resistance is blown out of proportion. To the
contrary, resistance is probably not being given its due, which brings us
to the second inherent problem of recognizing resistance identified at the
start of this section, i.e. identifying resistance early on and prior to the
appearance of clinical problems. Two principal factors contribute to our
inability to assess resistance adequately in the field. The first of these is
technological in nature; there simply are no good “cow-side” tests
available. Although a variety of assays have been developed (section
6.7), without exception they are poorly predictive and/or not conducive
to use in the field.
The second factor is more insidious as it is primarily philosophical in
nature. It is readily acknowledged that the time to combat resistance is
before it becomes entrenched. Theoretically, in early stages of resistance
development, while individual organisms are heterozygous for resistance
and potentially less fit than wild-type, susceptible organisms, it may be
possible to introduce strategies which allow for reversion to susceptibility
(Martin et al., 1988a; Maingi et al., 1990; Prichard, 1990a). Later in the
development of resistance, individuals become homozygous for the resis-
tant trait and as some evidence suggests, more fit (Kelly et al., 1978; Hall et
al., 1981b; Maingi et al., 1990), after which time reversion to susceptibility
is less likely (Maingi et al., 1990). Although some studies do not support
the fitness aspects of this hypothesis (MacLean et al., 1987; Waller et al.,
1989; Scott and Armour, 1991; Echevarria et al., 1993a), it is difficult to
interpret these data since in no case was the parent susceptible strain
compared to the resistant strain or the genetics of the populations known
with regard to resistance. To date, studies examining reversion to sus-
ceptibility have been overwhelmingly unsuccessful (Borgsteede and
Duyn, 1989; Martin, 1990; Jackson, 1993). Yet, almost without fail we
continue to look for resistance in the field by evaluating compounds at
recommended use level, a level which is characteristically well onto the
plateau of the dose-response curve and a level which could mask early,
subtle changes in resistant status. Instead, we should be studying the
response of parasites in the field at the shoulder of the dose-response
curve where small changes in resistance can be readily detected (Conder
et al., 1993; Shoop, 1993; Shoop et al., 1993b). This point is illustrated in
Figure 4, which is based on information from Conder et al. (1993). The
figure demonstrates moxidectin side-resistance for ivermectin in a jird
model, if a dose on the shoulder of the dose-response curve (i.e. 1.25 pg
per jird) is considered, but side-resistance would not be detected if a dose
on the plateau (i.e. 5 pg per jird) is used. Since the recommended use
level dose of moxidectin in ruminants is well onto the plateau of the
dose-response curve, side-resistance would not be detected at this dose as
was the case in studies reported by Craig et al. (1992), Oosthuizen and
Erasmus (1993), Pankavich et al. (1992), Pomroy et al. (1992), and Watson
et al. (1993) or would have been only marginally evident (Pomroy and
Whelan, 1993). It should be clear that two distinct issues are being confused
as one; by testing at use level in the field we are asking whether the drug in
question continues to have therapeutic efficacy regardless of resistance
status, not whether resistance to that drug is in place. Hence, resistance is
constantly being underestimated and strategies to limit or control resistance
are not implemented in a timely, potentially effective manner.
100-
m
.-E
.-
E
E
-
r 60-
u)
a
L
0
c
C
0
0 00-
i
r
0
0
40-
2
a
0
Ti
a
p 20-
c
c
a
e
2
0 ' I I I 1 I I
0.16 0.31 0.63 1.25 2.5 5

Dose of moxidectin (pgljird)
Figure 4 Dose-response curves for moxidectin against an ivermectin-susceptible
strain of Haemonchus contortus (0)and against an ivermectin-resistant strain of
H. contortus (0)in the jird.
6.4. Continuing Spread of Resistance
In spite of our inadequate efforts to identify resistance in the field as

outlined above, it is clear that resistance in nematodes is expanding
chemically, taxonomically, geographically, and clinically. Chemically,
each class of antinematodal drugs has enjoyed a short grace period before
resistance was identified, but no class of drugs currently available remains
without resistance in the field. Ivermectin, which was introduced in the
1980s as the first entry in the newest class of broad-spectrum antinematodal
drugs, generated resistance in H. contortus in the field in as few as three
treatments (Van Wyk et al., 1989a). Not only is the list of drug classes for
which resistance in nematodes has been identified continuing to grow, but
taxonomically the list of nematode species exhibiting resistance and the
range of hosts harboring resistant populations is also continuing to expand.
For example, early on resistance to levamisole was detected for Tricho-
strongylus spp., Ostertagia spp. and H . contortus in sheep and goats; only
later did resistance develop to other parasites in cattle (Geerts et al., 1987;
Williams, 1991; Williams et al., 1991), swine (Bjarn et al., 1989, 1990;
Roepstorff et al., 1987), and other hosts (Isaza et al., 1987; Malan et al.,
1988). In the case of the benzimidazoles, resistance has been identified in
nematodes from at least eight genera (Haemonchus, Ostertagia, Tricho-
strongylus, Nematodirus, Cooperia, Oesophagostomum, Chabertia, and
Strongyloides) in sheep and goats, four genera (Cyathostomum, Cylico-
cyclus, Cylicocostephanus and Strongylus) in horses, four genera
(Haemonchus, Ostertagia, Trichostrongylus and Cooperia) in cattle, one
genus (Ascaris) in swine, and still more in other host species. In addition,
there are now cases of single nematode species at a particular site being
resistant to all available classes of modern broad-spectrum anthelmintics
(McKenna et al., 1990; Watson and Hosking, 1990; Pomroy et al., 1992;
Varady et al., 1993) or nematodes of up to six genera being resistant to a
single drug on one property (McKenna, 1989).
Geographically, the spread of resistance is extending into areas such as
Europe and North America which were previously thought to be relatively
immune to problems of resistance, due to the combination of climate and
husbandry practices. The number of reports of anthelmintic resistance in
nematodes continues to grow dramatically, and there are indications that
we are only beginning to see the extent of resistance. For example, in a
survey of sheep and goats in the United States (Miller and Craig, 1988), it
was found that for each of the three classes of modem broad-spectrum
antinematodal drugs 25-53% of the sheep flocks examined harbored
nematode populations which were resistant, and nematodes in 57-75% of
goat flocks were resistant to benzimidazoles or levamisole. Although no
ivennectin resistance was present in goat flocks at that time, it has since
been identified in goats in the study area (Craig and Miller, 1990). Besides
expansion of resistance into geographic areas previously not recognized for
resistance, it is evident that resistance continues to expand on the local level.
A final proof of the expansion of resistance is the growing number of
clinical reports relating to resistance. Perhaps the most dramatic evidence
of this was reported by Van Wyk et al. (1988a, b) where resistance in some
regions of South Africa has got so completely out of hand that livestock
producers whose families have farmed for generations are getting out of
business due to the failure of all available anthelmintics to control
nematode disease in their animals.
6.5. Factors Contributing to Resistance
In most listings of contributors to resistance, frequent dosing heads the list.

Studies done on nematodes (Round et al., 1974; Barton, 1980, 1983;
Martin et al., 1982, 1984) clearly demonstrate that frequent dosing selects
for resistance more strongly than less frequent dosing regimes. In addition,
anecdotal evidence for-the role of frequent dosing in the generation of
resistant nematodes is that resistance has appeared most rapidly and has
been most pronounced in regions of South Africa, Australia and South
America where climatic conditions permit almost continuous exposure to
reinfection and acquisition of heavy nematode burdens, and where, as a
result, frequent dosing has been the cornerstone of control efforts. Even in
temperate climates frequent dosing is the rule as shown by a survey of
sheep farmers in the United States (Reinemeyer et al., 1992) where nearly
60%of ewes and rams and approximately,20% of lambs were dosed four or
more times a year. Frequent dosing is particularly troublesome when
dosing intervals are shorter than the prepatent period, since only resistant
worms are allowed to reach patency and contribute to the subsequent gene
pool; continued use of this practice for goats in New Zealand has resulted
in a high incidence of benzimidazole resistance and emergence of multiple
resistance (Jackson, 1993).
Underdosing is another major contributor in selecting for resistance.
Doses high enough to kill individuals heterozygous for resistance render
this trait in effect recessive, whereas subtherapeutic doses make it effec-
tively dominant (Roush and McKenzie, 1987). By using subtherapeutic
doses, marginally resistant heterozygous individuals are allowed to survive
and contribute resistant genes to subsequent populations. The example
cited earlier (Besier and Hopkins, 1988), where 86% of the farmers asked
to guess the weight of sheep (common practice in the field) would have
underdosed, demonstrates how frequently underdosing can occur. In
addition, animals are often further underdosed by using lower than
recommended doses to save money. Another potential contributor to under-
dosing is where a single host harbors different types of parasites (i.e.
.nematodes and arthropods) with different susceptibilities to a drug and
the drug is used at the dose sufficient for the more sensitive organism(s)
on a spot basis. A good example of this is the use of ivermectin to control
Hypoderma spp., since ivermectin is active against this parasite at doses
well below those recommended for nematodes. Inadequate attention to
calibration or condition of dosing equipment can also result in delivery
of less than the desired dose. The extreme case of underdosing is where an
animal is inadvertently not dosed.
It is generally believed that alternation of drug classes between but not
within parasite generations or on an annual basis can prevent or slow
development of resistance (Prichard et al., 1980). However, in practice,
farmers tend to use a single drug until it no longer works. As an illustration
of this point, Reinemeyer et al. (1992) surveyed sheep farmers in the
United States and found this to be the case, since approximately one out
of two herds were being dosed with a single anthelmintic until it failed.
Unfortunately, when farmers do alternate, they often tend to do so within a
season rather than on a yearly basis (> 50% of respondents in the above
survey) and/or to use drugs within the same class. In the former case,
multiple resistance may be generated, whereas in the latter, selection for
resistance will continue unchecked, since all compounds within a class
utilize the same mode of action.
Management practices can contribute in a variety of ways to the genera-
tion of resistance. Clearly, relying exclusively on drugs to control nema-
tode infections is a poor management practice that can and often does lead
to resistance. Introducing new animals that may harbor populations of
resistant parasites onto a farm without appropriate quarantine, disease
monitoring, and treatment can rapidly result in resistance problems.
Anthelmintic dosing schemes which ignore the particular characteristics
of development and transmission of the parasites in question, particularly
the availability of susceptible free-living stages in the environment which
can contribute to subsequent infection, may result in enhancing resistance.
An example of this latter point for H. contortus is where sheep in temperate
climates are dosed during the periparturient period. Since the parasite’s
free-living stages are susceptible to cold, they are greatly reduced on
pasture at this time, leaving the major source of subsequent infection to
be the progeny of those worms which are resistant and survive treatment
(Taylor, 1990a; Sykes et al., 1992). Dosing animals and moving them to
“clean” or minimally contaminated pasture has long been advocated;
however, this practice could contribute to resistance development
because, as in the above example, those worms which are resistant and
survive treatment would be the major contributors to subsequent infection
(Le Jambre, 1978; Taylor and Hunt, 1989; Smith, 1990). Commingling
host species such as sheep and goats as noted above can lead to cross-
transmission of resistant parasites from one host (goats) where resistance is
more readily generated to another (sheep) in which resistance may be less
likely to occur. In addition, ignoring the pharmacokinetic differences
between species as in the case of sheep and goats can result in underdosing
(Coles et al., 1989a; Sangster et al., 1991a) and generation of resistance for
goats treated at doses recommended for sheep. Likewise, pharmacokinetic
differences between breeds of the same species or even within a breed
under diverse management schemes can affect dose requirements and can
contribute to resistance.
6.6. Mechanisms of Resistance
Drug resistance generally results from decreased drug uptake, increased

metabolism of the drug, or changes at the drug-receptor site (Prichard,
1990b; Roush, 1990). To date, there is little evidence to indicate that

changes either in drug transport or drug metabolism in nematodes play
any significant role in resistance to antinematodal drugs (Prichard, 1990b),
but as outlined below, changes at the drug-receptor site may play a key
role. Although we are a long way from understanding the mechanisms of
resistance in most cases for anthelmintics, progress is being made. The
mechanisms of resistance for the benzimidazoles are best understood, at
this time.
Benzimidazoles are generally believed to work by interfering with
tubulin polymerization into microtubules (reviewed by Lacey, 1990). It
is worth noting that phenothiazine appears to share the benzimidazole
mode of action (Rew and Fetterer, 1986) and, as suggested by Coles
(1988) and Drudge et al. (1990), its use may have preselected some
nematode populations for benzimidazole resistance. Consistent with this
mode of action, it appears that resistance to the benzimidazoles is due to
the loss of high affinity receptor binding sites resulting from a change in P-
tubulin isotype pattern. This conclusion is based on a variety of studies,
only a few of which will be detailed here. Lacey and Snowdon (1988) using
radiolabeled mebendazole in a binding assay found that partially purified
tubulin from mebendazole-sensitive strains of T. colubrijormis and H .
contortus were bound at significantly higher levels than was tubulin from
resistant strains of the two parasites, despite essentially identical tubulin
levels. This suggested a reduction in binding affinity and Lubega and
Prichard (1991) showed that high affinity binding sites were indeed
decreased in resistant populations and that the number and frequency of
P-tubulin restriction fragments decreased with selection for resistance.
Roos et al. (1990) extended these observations. Using restriction mapping
with cloned a-and P-tubulin genes from H. contortus as probes they found
no differences in a-tubulin from benzimidazole-susceptible and -resistant
populations. However, they identified between two and six P-tubulin
fragments, designated isotype 1 and including genes isolated by Roos et
al. (1990) and Geary et al. (1992a), in susceptible worm populations, but
only one to two of the isotype 1 P-tubulin fragments were present in
resistant worms. There were always fewer fragments in resistant than
susceptible populations when the same restriction enzyme was used for
each population. Similar results were found for isotype 1 with regard to
susceptible and resistant strains of T. colubrijormis (Le Jambre, 1990).
Further, Roos et al. (1990) found that a 9 kb fragment was always present
in resistant populations and in some susceptible populations. Based on this
information, they postulated that benzimidazole treatment selected for a
pre-existing P-tubulin in susceptible populations. To test this hypothesis,
they used a benzimidazole-susceptible laboratory strain that predated the
introduction of the benzimidazoles (it is unclear whether this strain had
previously been exposed to phenothiazine, which may preselect for benzi-

midazole resistance as noted above), selected this strain through two
generations for benzimidazole resistance, and then examined both the
susceptible parent and derived resistant strain using restriction mapping.
Although one to five P-tubulin fragments, including the 9 kb fragment,
were found in the susceptible population, essentially a single 9 kb fragment
was obsefved for individuals from the resistant populations. Further selec-
tion of this resistant population (Kwa et al., 1993) produced a single 9 kb
fragment and demonstrated that higher levels of benzimidazole resistance
could not be correlated with additional change in P-tubulin, suggesting
othet mechanisms play a role. In addition, Kwa et al. (1994), using an
allele-specific PCR, have shown that all benzimidazole-resistant isolates of
H. contortus and T . colubriformis examined have a Phe to Tyr mutation at
amino acid 200 in isotype 1 P-tubulin. It remains to be seen whether this
amino acid substitution is the actual cause of resistance or simply a
genetically linked factor.
A second P-tubulin isotype, designated isotype 2 and including genes
isolated by Geary et al. (1992a), when used as a probe in restriction
mapping analysis by Geary et al. (1992a) could not distinguish on the
basis of banding between resistant and susceptible populations. However,
Kwa et al. (1993) using an isotope 2 probe found little difference in
banding patterns between susceptible and the less resistant populations
of the same selected lines used for isotype 1 as noted above, which
exhibited ED50 in an egg hatch assay similar to ED50 of the resistant
strains used by Geary et al. (1992a), but in the most resistant population
(the increase in resistance in this over less resistant populations could not
be correlated with a change in isotype l), no isotype 2 genes were detected.
Further, they demonstrated isotype 2 genes were present in eggs, third-
stage larvae, and adults of the fully susceptible parent but not the highly
selected resistant population, and that a field isolate with a high level of
resistance comparable to the highly selected strain had no or a very low
amount of isotype 2. Since both the highly selected and field isolate are
apparently normal viable populations, isotype 2 is probably redundant. Le
Jambre (1993b) summarized similar results for T. colubriformis, which
suggest this parasite also deletes the gene encoding isotype 2 in highly
selected resistant individuals. These data appear to support the conclusion
that benzimidazole pressure selects for changes in P-tubulin isotype
patterns through at least two mechanisms, i.e. (1) selection of a pre-
existing 9 kb isotype 1 P-tubulin gene (or a genetically closely linked
factor) with reduced affinity for benzimidazoles and (2) elimination at
higher resistance levels of individuals with isotype 2 genes. Kwa et al.
(1993) suggest that isotype 1 and 2 map to different loci and isotype 1 has
multiple alleles, and although this remains to be proven, other reports
indicate benzimidazole resistance in trichostrongylid nematodes is poly-

genic (Le Jambre et al., 1979b; Herlich et af., 1981; Martin et af., 1988b).
In addition, evidence that multiple P-tubulin isotypes can differ in degree
of benzimidazole resistance comes from work on the free-living nematode
C. elegans (Driscoll et al., 1989) and fungi (May et al., 1985; Orbach et
al., 1986; Sheir-Neiss et af., 1978; Steams and Botstein, 1988) where
mutations conferring benzimidazole resistance map to P-tubulin genes.
As indicated earlier, less is known about the molecular mechanisms of
resistance for the other classes of broad-spectrum anthelmintics, although
the data suggest that, as for the benzimidazoles, multiple mechanisms are
in place for ivermectin and levamisole. Resistance to ivermectin in the
free-living nematode C. elegans appears to be polygenic as it has been
shown to map to mutations in any of several (> 23) genes distributed over
at least five chromosomes (Day et al., 1989 as reported by Le Jambre,
1990). Since there is no reason that drug resistance should be due to a
single mechanism within a species, or even among different species, the
data from C. elegans are not surprising. Gill et af. (Gill, J.H., Redwin,
J.M. and Lacey, E., personal communication) using motility and larval
development assays showed different phenotypic responses in ivermectin-
resistant strains of H. contortus and T. colubriformis, both between and
within species. In addition, Scott (1989) has shown that two mechanisms
are effective in producing abamectin-resistance in one strain of Musca
domestica. The two mechanisms are decreased cuticular penetration of
the drug and increased drug metabolism via the mixed-function oxidase
system as a result of mutations on chromosomes 3 or 2, respectively.
However, in a second resistant strain of M . domestica, Scott et af.
(199 1b) were unable to demonstrate a metabolism component associated
with the resistance and suggested the high level of resistance was not
typical of a decreased cuticular penetration mechanism. Hence, they
suggested an altered target site sensitivity. Grant et al. (Grant, W.N. and
Hunt, P.W., personal communication) demonstrated that decreased drug
uptake may contribute to ivermectin resistance in nematodes, since all
G. elegans resistant to ivermectin are amphid defective, whereas sensitive
strains are all normal with respect to amphids. This, combined with the
supposition by Martin and Kusel(l992) that ivermectin can transport along
but not across membranes, could implicate amphids in a major uptake
function which is reduced or absent in the case of resistant organisms,
because of defective amphids.
In the case of levamisole resistance in parasitic nematodes, multiple
mechanisms also seem to be in effect, as Martin and McKenzie (1990)
report levamisole resistance in one strain of T. colubriformis appears to be
sex-linked recessive in nature, whereas Sangster (1990) suggests multigenic
inheritance of levamisole resistance occurs in a strain of H. contortus.
Resistance to this drug, in at least some strains of H. contortus and T.

colubriformis, appears to be the result of a reduction in the number of
levamisole receptors or in their affinity (Sangster et al., 1988, 1991b). This
supposition is consistent with studies on C. elegans in which levamisole-
resistant mutants lack acetylcholine receptors (Lewis et al., 1980) and
differential binding of levamisole and related drugs between susceptible
and resistant strains has been documented (Lewis et al., 1987). Clearly, a
great deal more work is needed if we are to understand the molecular basis
for resistance in parasitic nematodes.
6.7. Monitoring/Reporting of Resistance
A wide variety of assays (reviewed by Presidente, 1985; Johansen, 1989;

Taylor and Hunt, 1989; Coles, 1990a) are available to monitor resistance.
Unfortunately, as noted earlier, these are either poorly predictive and/or are
not conducive to “cow-side” use. Although controlled efficacy tests in vivo
have been considered the definitive means to determine whether resistance
is present in a population, this approach is prohibitively expensive and time
consuming. In addition, efficacy tests, as generally run, are incapable of
detecting resistance until it is well established. Historically, fecal egg count
reduction assays (Presidente, 1985; Webb and Ottaway, 1986; Vizard and
Wallace, 1987; Martin et al., 1989; McKenna, 1990b,c), which compare
pre-treatment egg count levels to those following treatment and which are
predictive for all anthelmintic classes, species of nematodes which shed
eggs in the feces, and hosts, have been widely used as a measure of
treatment success; they continue to be a mainstay due to their relative
ease and versatility. An Australian Working Party on Anthelmintic
Resistance (Anonymous, 1989) has recommended refinements, including
(1) the use of arithmetic as opposed to geometric means, (2) requiring that
mean pre-treatment egg counts are >150 g-’ of feces, (3) collection of
post-treatment (PT) samples at 10-14 days PT (longer may be required for
ivermectin; Jackson, 1993), and (4) requiring that egg reductions be 1 95%
with the lower 95% confidence level being greater than 90%. These
refinements have enhanced the predictive nature of egg count reduction
assays with regard to identifying resistant populations. In fact, the World
Association for the Advancement of Veterinary Parasitology has recom-
mended that fecal egg count reduction and egg hatch assays (the latter will
be discussed below) be used in monitoring for resistance in nematodes
(Coles et al., 1992). Nevertheless, because of poor sensitivity, fecal egg
count reduction assays are capable of detecting only high levels of
resistance. Resistance for minor species in the population mix may be
missed using this technique when major species vastly outnumber them
(Presidente, 1985; Martin et al., 1989; Beveridge et al., 1990). In addition,

for some parasites (e.g. Nematodirus spp.) fecal egg counts may not be
reliable indicators of infection (Black, 1964; Kingsbury, 1965; McKenna,
1981; Chalmers, 1985; Martin et al., 1985) and hence may be of ques-
tionable value in assessing resistance (Obendorf et al., 1991). Larval culture
can be used in conjunction with fecal egg count reduction assays to identify
which parasites are resistant, but caution must be used because highly
fecund nematode species may mask species with low fecundity (West et
al., 1989) and species which have low mortality in culture may be over-
estimated relative to those that suffer high mortality in culture (Dobson et
al., 1992). Reinecke et al. (1991) described a first-stage larval reduction test
. as an interesting alternative to the fecal egg count reduction assay. In this
case, first-stage larval populations (numbers and generic composition)
which develop from eggs recovered from treated animals are compared to
those from non-treated animals. The main advantage of this assay is that it
appears to be better able to detect minor contributors to the population.
A number of in vitro assays, based on anthelmintic effects on normal
physiological processes such as development, growth, and/or movement
and widely applicable to strongylid species (excluding nematode species
such as N . spathiger where the eggs do not hatch rapidly), have been
developed to support egg count reduction assays. In each case, they are
not suitable for field use and have little utility where metabolic activation is
required. They include egg hatch, larval development, larval paralysis,
motility, and larval migration assays. The egg hatch assay as described
by various authors (Le Jambre et al., 1970; Le Jambre, 1976; Coles and
Simpkin, 1977; Whitlock et al., 1980; Boersema et al., 1982; Cawthorne
and Whitehead, 1983; Smith-Buijs and Borgsteede, 1986) is based on the
ovicidal activity of the benzimidazoles and compares the ability of fresh,
undeveloped eggs from the population in question to embryonate and hatch
following exposure at various concentrations of anthelmintic to that of
similarly treated eggs from a population known to be susceptible to the
anthelmintic. The assay works well for identifying resistance to the benzi-
. midazoles (Dobson et al. (1986) adapted the assay for levamisole where the
drug actually paralyzes the first-stage larva in the egg), but it suffers from its
limited utility across the complete spectrum of available antinematodal
drugs, in working only for nematode species that hatch rapidly, from the
requirement for transporting eggs from the field to the laboratory while
preventing development of the eggs, and in a lack of correlation between
relative levels of resistance in the assay versus the field. The problem of
getting undeveloped eggs to the laboratory can be dealt with by trans-
porting them on ice (Smith-Buijs and Borgsteede, 1986) or anaerobically
if longer periods (up to 7 days) are required for transport or until assay
(Hunt and Taylor, 1989).
Larval development assays (Coles et al., 1988; Giordano et al., 1988;

Stringfellow, 1988; Lacey et al., 1990; Taylor, 1990b; Hubert and Kerboeuf,
1992) examine the influence of anthelmintics at differing concentrations on
subsequent development of eggs or first-stage larvae to infective third-
stage larvae or, in the case of Stringfellow (1988), from exsheathed
third-stage to fourth-stage larvae. These assays, besides having utility for
detecting resistance to all three classes of modem broad-spectrum anthel-
mintics, have the added benefits of not requiring undeveloped eggs and of
more easily allowing identification of the resistant species. The utilization
of an agar matrix containing the drug by Lacey et al. (1990) apparently
enhances the utility of the assay for compounds such as ivermectin where
solubility in liquid media as used in the other assays of this type may be a
problem. On the downside, substantially more time is needed to obtain
results with these than other in vitro assays, resistant species with lower
reproduct.ive rates may be masked by overwhelming numbers of suscep-
tible species in mixed populations, reading this type of assay is tedious and
subject to interpretation, and again these are laboratory assays. Taylor
and Hunt (1989) reported on an adult development test where early
fourth-stage larvae in culture are treated and allowed to develop to late-
fourth-stage larvae or adults, and although they indicated that differential
development was detectable between resistant and susceptible populations
of 0. circumcincta and H . contortus, no further details were provided.
Three types of assays based on the ability of larvae (or adults) to move
have been developed. The first of these is the larval paralysis test (Martin
and Le Jambre, 1979) in which worms are observed visually for their
ability to move following anthelmintic treatment. This is a very subjective
laboratory assay, its reliability has been questioned by some authors
(Barton, 1983; Boersema, 1983b), and it is very labor intensive. In addi-
tion, larval age may influence the results for the paralysis assay (Geerts et
al., 1989). Over the last decade, paralysis assays have been developed
which are more objective and better quantified. These have been desig-
nated motility assays and are described by various authors (Folz et al.,
1987; Coles et al., 1989b; Gill et al., 1991). The most objective and
sensitive of these assays is that described by Folz et al. (1987) which
utilizes a micromotility meter (developed and described by Bennett and
Pax, 1986) to detect changes in motility patterns of treated compared to
non-treated worms. These motility assays are useful for detecting resis-
tance to benzimidazoles or macrocyclic lactones but suffer from lack of
utility for detecting resistance to levamisole (Sangster et al., 1988; Coles et
al., 1989b). In addition, the sophisticated equipment utilized for these
assays renders them even more confined to the laboratory. Another move-
ment assay is a migration inhibition assay (Wagland et al., 1992; Rothwell
and Sangster, 1993) based on the ability of treated larvae to move from one
chamber through a mesh sieve into a second chamber. When culture-

derived late third-stage or early fourth-stage larvae of H . contortus are
used, this assay is usefulfor detecting resistance to each of the three classes
of modem broad-spectrum anthelmintics, as well as the salicylanilides for
which none of the other in vitro assays has proven useful. By way of
explanation, Rothwell and Sangster (1993) hypothesize that although
closantel is capable of killing early (free-living) stages of H. contortus in
vitro, it does so via transcuticular uptake, and this action may bypass the
cellular mechanism, thus preventing expression of closantel resistance. In
their assay, on the other hand, the test larvae mostly have reached the
fourth stage, which begins to feed and ,so takes up closantel bound to
. protein in the assay medium, mimicking the situation in vivo for this
hematophagous parasite and allowing resistance to be shown. Alterna-
tively, they suggest that the resistance mechanism may not develop until
parasitic stages develop. Unfortunately, the assay's complexity may limit
its usefulness.
In addition to the physiologically based in vitro assays, biochemical- and
genetic-based in vitro assays have been, or could be, used to examine
resistance. Colorimetric (Sutherland et al., 1988, 1989) or paralysis
(Sutherland and Lee, 1990) tests, based on the level of non-specific
esterases or acetylcholinesterases which are elevated in benzimidazole-
resistant compared to susceptible worms, are examples of biochemical
assays. Others include tubulin-binding (Lacey, 1985; Lacey and Snowdon,
1988) and tubulin-polymerization (Lacey and Richard, 1986) assays as
measures of benzimidazole resistance (in benzimidazole-resistant strains
these drugs are unable to bind to tubulin and hence fail to interfere with
tubulin polymerization). At the present time, genetic assays are limited to
the benzimidazoles where resistant populations have been identified using
cloned P-tubulin probes with restriction mapping (Le Jambre, 1990, 1993b;
Roos et al., 1990) or isoenzyme analysis using isoelectric focusing
(Sutherland et al., 1988). Echevarria et al. (1992) also showed that iso-
enzyme analysis could be useful in identifying ivermectin-resistant strains,
but not if benzimidazole resistance was also present as the difference
between ivermectin-sensitive and -resistant strains is masked by elevated
enzyme levels due to the benzimidazole resistance. With the possible
exception of the colorimetric assays which have proven useful in a simpli-
fied format in the field for identifying organophosphate and carbamate
insecticide resistance in aphids (Sawicki et al., 1978), none of the available
biochemical- or genetic-based approaches is conducive to field use, due to
expense and technical requirements. Being very specific in nature, most of
these assays are also not useful in assessing resistance to the spectrum of
drugs available.
Other components in the battery of assays available to assess resistance
are the target parasite/rodent models which have utility for all the modem
broad-spectrum antinematodal drugs. The only models of this type used
to date to assess resistance of suspect field strains compared to strains
known to be susceptible to the drugs in question or to assess crosshide
resistance between drugs are jirds infected with T. colubriformis (Ostlind
et al., 1990) or H . contortus (Conder et al., 1991, 1993; Duwel, 1991) or
guinea pigs infected with T. colubriformis (Kelly et al., 1981a) or H .
contortus (Rolfe, 1990). Clearly, although in vivo rodent models are
available for all classes of modem anthelmintics, they have not been
developed for the majority of parasites of interest, they are expensive
artd labor intensive relative to in vitro assays, they are limited to the
laboratory, and they require the use of animals. Nevertheless, they
provide some unique advantages over both egg reduction and in vitro
assays which have not been fully exploited to date. Whereas egg reduc-
tion tests do not fully assess the survival of worms following anthelmintic
treatment and even spot examination of animals for actual worm survival
in target hosts by necropsy techniques is prohibitively expensive, parallel
studies in rodent models can fully assess worm survival. Not only is it
possible to determine worm survival following treatment in rodent mod-
els, but this can be done in a systematic and cost-effective manner which
first identifies the shoulder of the dose-response curve (the most sensitive
point to detect a change in susceptibility) for a susceptible strain and
subsequently compares the response for the suspect strain. Hence, resis-
tance and not efficacy is being measured. The utility of this approach has
been demonstrated in rodents (Conder et al., 1993) and substantiated by
target host studies (Shoop et al., 1993b). In addition, the resistance
factors for resistant strains relative to susceptible strains for these in
vivo models more closely parallel the situation in the field than do the
in vitro assays.
It is obvious that we need to develop better means to identify resistance
in the field. It is equally obvious that the absence of a good “cow-side”
assay for resistance should not be an excuse for inadequately monitoring
resistance development. We need to use the tools at-hand more wisely.
Clearly, it is not acceptable to examine drugs in the field at use level as a
measure of incipient resistance; using this approach, resistance will only be
recognized when it is beyond the point where anything can be done to
control it. Since the only assays currently available which are capable of
identifying resistance early in the developmental process are those in the
laboratory, it is imperative that surveillance programs utilizing these assays
be implemented. The program initiated in England by Coles (1990b) and
the “Wormtest” campaign in Australia are reasonable examples of such a
surveillance program.
6.8. Strategies to Limit Resistance Development
Since currently available assays are impractical for field use and/or are
unable to detect early resistance in the field, and it is difficult or impossible
to render populations with high degrees of resistance susceptible again, it
makes sense to strive to prevent resistance from developing rather than
managing resistance once it is in place (Roush, 1990). With this in mind, a
number of strategies have been suggested to limit the development of
resistance. It should be recognized that any strategy must fit the particular
characteristics (parasite and host biology, environment, epidemiology, etc.)
of the target population, and hence, strategies will vary considerably
. between or even within regions.
Perhaps the most widely advocated practice to prevent resistance deve-
lopment is to limit the number of anthelmintic treatments. By reducing
exposure to drug, selection pressure can be minimized. To do this, how-
ever, it is imperative that the parasites present and the epidemiology of
helminth disease locally be known, so dosing schedules can be strategically
timed and integrated with other management practices to prevent disease,
limit parasite proliferation, and optimize production. History has taught
us that, in fact, only rarely are anthelmintics used strategically. Educa-
tion programs need to stress that although alternative nematode control
strategies relying less heavily on anthelmintics may be more costly than
reliance solely on drugs, once resistance develops there are fewer options
to control these parasites and greater costs due to reduced efficiency or
profits (Roush, 1990). A corollary of limited, strategic use of anthelmintics
is to use only a single class of anthelmintics annually or within a parasite
generation, so multiple (class) resistance is not generated, and to rotate
anthelmintic classes on a yearly basis to limit passage of resistance genes
(resistant to the previous class but susceptible to the alternate class of
ahthelmintic) early in the selection process while organisms are hetero-
zygous for the trait allowing for a reversion to susceptibility. In addition,
periodic assessment of resistance status should be a standard part of any
nematode control strategy utilizing anthelmintics. Use of any anthelmintic
should be discontinued if resistance to it is detected and subsequent
treatments should use a drug with a different mode of action.
A critical aspect of limiting resistance development, regardless of dosing
scheme, is the need to assure that an effective dose of a highly efficacious
drug be delivered, so that heterozygous resistant individuals are effectively
removed along with susceptible worms, i.e. animals are not underdosed.
Toward this end, animal weights should be accurately determined, not
guessed, and group dosing should be based on the heaviest animal. In
addition, label directions should be followed explicitly with regard to
dose, route, target parasites, target hosts, and expiration date.
Treatment followed by subsequent movement of animals to “clean” or

minimally contaminated pasture has long been advocated as a management
practice (Barger, 1978; Morley and Donald, 1980). In support of this
strategy, a 5-year study in Australia produced no evidence that it resulted
in more resistance than did permanent pasture (Waller et al., 1989) and a
mathematical model on resistance to 0. circumcincta suggested the dose
and move strategy would select less readily for resistance than permanent
pasture (Gettinby et al., 1990). This approach is severely limited by pasture
availability, a particular problem for the small producer, and it has been
questioned as an effective practice (Le Jambre, 1978; Taylor and Hunt,
1989; Smith, 1990) on the basis that resistant worms from the treated
animals would be the major contributors to reinfection. A number of
reports support the contention that this practice contributes to resistance
development (Vlassoff and Kettle, 1980; Cawthorne and Whitehead, 1983;
Martin er al., 1985; Taylor and Hunt, 1988; Martin, 1989). Obviously, the
advantage of reducing exposure to reinfection and the risk of selecting for
resistance must be considered in light of all other factors on a case-by-case
basis before dose and move strategies are used.
Preventing the introduction of resistant parasite strains with new animals
by quarantining, monitoring, and treating all replacement stock is a critical
management practice. For the reasons outlined earlier in this chapter,
commingling of species such as sheep and goats is generally inadvisable.
In some cases, removal of feces from pasture may be a useful management
tool (Herd, 1986b). Where practical, alternate grazing of livestock of
various species or immune status has been advocated (Hall et al., 1981c;
Eysker et al., 1983b, 1986; Bisset et al., 1988). Selectively deworming
animals from a population has also been proposed as a means of limiting
selection for anthelmintic resistance (Duncan and Love, 1991; Sykes et al.,
1992). For example, it has been demonstrated (Sykes et al., 1992) that most
pasture contamination (up to 70% of nematode larvae) comes from ewes
rather than lambs. Therefore, strategic treatment of ewes could reduce
pasture contamination to a level where fewer treatments would be needed
in lambs.
An interesting strategy proposed by Van Wyk and Van Schalkwyk
(1990) is the reintroduction of susceptible strains into areasherds where
resistance is a problem. This approach remains unproven, and it is unlikely
that resistant strains can be displaced in this manner unless there is a
significant fitness advantage for the susceptible strain, since the numbers
are decidedly in favor of the resistant parasites.
Narrow-spectrum drugs can be used effectively to control some species
which have developed resistance to the broad-spectrum anthelmintics, e.g.
H. contortus. This is well exemplified by the strategic use of closantel in
combination with broad-spectrum drugs as outlined by Dash (1986b) in
programs such as “Wormkill” in Australia, where local eradication of H .

contortus has been demonstrated (Barger et al., 1991). Unfortunately, this
and other similar programs are jeopardized by emerging resistance to
closantel and related narrow-spectrum anthelmintics (Van Wyk and
Gerber, 1980; Van Wyk et al., 1982, 1987; Van Wyk and Malan, 1988;
Rolfe et al., 1990).
The use of mixtures of two broad-spectrum drugs from different classes
(i.e. different modes of action) has been advocated (Anderson et al., 1988a;
Daly, 1990) as a means to delay resistance development. This approach has
been utilized for years with antibacterials (Curtis, 1985) and is currently
advocated for antimalarial therapy (Anonymous, 1984; Peters, 1984). It is
. based on the concept that it is unlikely any individual in the population will
carry resistance alleles for both classes if both are found at low frequency
in the population, i.e. before selection for either class has occurred. The use
of drug combinations to control nematodes is supported by studies in which
the drugs have been administered sequentially (Anderson et al., 1988a),
simultaneously (McKenna, 1990a), or in divided doses over 24 h (Sangster
et al., 1991a) and by simulation models (Martin, 1990; Smith, 1990).
Although optimally drug mixtures should be used before resistance to
either component of the mixture has arisen, it can be argued that this
approach has utility even where resistance is in place, as long as no
species in the parasite population to be treated is bi-resistant to the drugs
in the mixture (McKenna, 1990a). Compounds with dramatically different
persistence may be a problem with this approach since resistance may be
selected for the compound with a longer decay time, as the compound with
shorter persistence is not available to remove recruited individuals resistant
to the longer-persisting drug during its protracted decay. Thus, although the
use of a macrocyclic lactone as one component of a mixture may have the
advantage of less resistance currently being present in the field, compounds
in this class are known to have substantial persistence and hence may suffer
from this attribute in combinations. Sustained- or pulsed-release anthel-
mintics may also provide a useful tool in controlling resistant nematodes or
preventing their selection (Sangster et al., 1992), if used in a judicious
manner.
In recent years, mathematical models have been used to evaluate factors
contributing to resistance and/or strategies to limit its development (Martin
et al., 1984; Waller et al., 1985; Dobson et al., 1987; Gettinby, 1989;
Gettinby et al., 1989, 1990; Barnes and Dobson, 1990; Martin, 1990;
Martin and McKenzie, 1990; Smith, 1990; Echevarria et al., 1993b).
These models are powerful tools which can predict optimum strategies
combining judicious use of anthelmintics with other management practices
to minimize the impact of nematode parasitism on production while
prolonging the useful life of the limited anthelmintic arsenal. However,
caution should be exercised in using these models, since they require field
confirmation and are dependent on accurate information regarding nema-

tode biology and epidemiology, genetics of resistance, acquired immunity
in the host, current management practices and options, and environmental
factors, all of which may vary from location to location and over time.
Although it is currently not possible to incorporate nematode-resistant
host libes or nematode vaccines (with the exception of attenuated vaccines
for lungworms) into control strategies, there is reason for optimism that
these approaches will become useful, albeit not in the immediate future.
Likewise, approaches to controlling pasture contamination through the use
of nematophagous fungi, Bacillus thuringiensis toxins, or growth regu-
lators offer promise. Clearly, any control measure which reduces our
reliance on chemotherapy will also diminish the rate at which resistance
develops and its subsequent impact.
6.9. Epilogue
Never has the arsenal of antinematodal drugs been more formidable than
today. The three classes of modem broad-spectrum anthelmintics provide
an unprecedented spectrum of activity (including not only nematodes but
also flatworms and ectoparasites), a wide range of delivery options, and the
opportunity to develop and use a variety of treatment strategies. Although
alternatives to chemotherapeutic control of nematodes are being explored
and showing promise, it is unlikely that any of these approaches will be
broadly available in the near future. This combined with the fact that
anthelmintic resistance is a reality dictates prudent use of anthelmintics,
implementation of integrated control strategies, and early recognition of
and appropriate response to resistance development.
ACKNOWLEDGEMENTS
The authors thank Dr David Thompson for providing Figure 1, Dr Stephen

J. Nelson for preparing Figures 2 and 3, and Ms Wendy L. Buckner for
typing the manuscript. For assistance in compiling data on regulatory status
of drugs and on approval of efficacy claims, the authors are grateful to Dr
James L. Cox and Ms Mary E. Doscher.
NOTE ADDED IN PROOF
Benzimidazole resistance in T. colubriformis in goats from the United

Kingdom (Hunt et al., 1994) and in Ostergia spp. and Cooperia spp. in
sheep from Greece which were imported from France and the United
Kingdom (Himonas and Papadopoulos, 1994) has been reported.
REFERENCES
Andersen, F.L. and Christofferson, P.V. (1973). Efficacy of haloxon and thiaben-
dazole against gastrointestinal nematodes in sheep and goats in the Edwards
Plateau area of Texas. American Journal of Veterinary Research 34,1395-1398.
Anderson, N. ( 1977). The efficiency of levamisole, thiabendazole and fenbenda-
zole against naturally acquired infections of Ostertagia ostertagi in cattle.
* Research in Veterinary Science 23, 298-302.
Anderson, N. and Lord, V. (1979). Anthelmintic efficiency of oxfendazole, fenben-
dazole and levamisole against naturally acquired infections of Ostertagia
ostertagi and Trichostrongylus axei in cattle. Australian Veterinary Journal
55, 158-162.
Anderson, N., Martin, P.J. and Jarrett, R.G. (1988a). Mixtures of anthelmintics: a
strategy against resistance. Australian Veterinary Journal 65, 62-64.
Anderson, N., Martin, P.J., Jarrett, R.G., Brown, T.H. and Miller, D.W. (1988b).
Sequential development of resistance to thiabendazole and levamisole in
nematodes of sheep. International Journal for Parasitology 18, 243-249.
Andrews, S.J., Ferrari, M.M., Pow, J.D.E. and Lancaster, M.B. (1993). Nematode
egg output and plasma concentration of ivermectin after its administration to red
deer (Cervus elaphus elaphus). Veterinary Record 132, 161-163.
Anonymous, A.A. (1984). Advances in malaria chemotherapy. World Health
Organization Technical Report Series No. 7 11.
Anonymous, A.A. (1986). Directorate of Veterinary Services, South Africa:
Laboratory Services Monthly Reports: January, p. 4.
Anonymous, A.A. (1989). Report of the Working Party for the Animal Health
Committee of the Standing Committee on Agriculture. SCA Technical Report
Series No. 28, Anthelmintic Resistance, Commonwealth Scientific and Industrial
Research Organization, Australia.
Anonymous, A.A. ( 1992). Anthelmintic resistance: Learning from the Australian
experience. Veterinary Record 130, 362-363.
Arena, J.P., Liu, K.K., Paress, P.S. and Cully, D.F. (1991). Avermectin-sensitive
chloride currents induced by Caenorhabditis elegans RNA in Xenopus oocytes.
Uolecular Pharmacology 40,368-374.
Arena, J.P., Liu, K.K., Paress, P.S., Schaeffer, J.M. and Cully, D.F. (1992).
Expression of a glutamate-activated chloride current in Xenopus oocytes injected
with Caenorhabditis elegans RNA: evidence for modulation by avermectin.
Molecular Brain Research 15, 339-348.
Badger, S.B. and McKenna, P.B. (19%). Resistance to ivermectin in a field strain
of Ostertagia spp. in goats. New Zealand Veterinary Journal 38, 72-74.
Baker, R. and Swain, C.J. (1989). Ivermectin - a drug for all seasons. Chemistry
in Britain 25, 692-696.
Baker, N.F., Miller, J.E., Madigan, J.E., Fulton, R. and Seward, R.L. (1984).
Anthelmintic action of ivermectin, oxibendazole and pyrantel pamoate against
thiabendazole-resistant small strongyles of horses. Equine Practice 6, 8, 10-12,
1 4 1 5 , 17-19.
Bard, J.H. (1972). “Anthelmintic Index”. Commonwealth Agricultural Bureaux,

Slough.
Barger, I.A. (1978). Grazing management and control of parasites in sheep. In
“The Epidemiology and Control of Gastrointestinal Parasites of Sheep in
Australia” (A.D. Donald, W.H. Southcott and J.K. Dineen, eds), pp. 53-63.
CSIRO, Melbourne.
Barger, LA. (1993). Control of gastrointestinal nematodes in Australia in the 21st
century. Veterinary Parasitology 46, 23-32.
Barger, I.A. and Lisle, K.A. (1979). Benzimidazole resistance in small strongyles
of horses. Australian Veterinary Journal 55, 594-595.
Barger, I.A., Hall, E. and Dash, K.M. (1991). Local eradication of Haemonchus
contortus using closantel. Australian Veterinary Journal 68, 347-348.
Barnes, E.H. and Dobson, R.J. (1990). Population dynamics of Trichostrongylus
colubrijormis in sheep: computer model to simulate grazing systems and the
evolution of anthelmintic resistance. International Journal for Parasitology 20,
823-83 1.
Barragry, T.B. (1987). A review of the pharmacology and clinical uses of
ivermectin. Canadian Veterinary Journal 28, 5 12-5 17.
Barton, N.J. (1980). Emergence of Haemonchus contortus resistant to thia-
bendazole. Australian Veterinary Journal 56, 46-47.
Barton, N.J. (1983). Development of anthelmintic resistance in nematodes from
sheep in Australia subjected to different treatment frequencies. International
Journal for Parasitology 13, 125-1 32.
Barton, N.J., Trainor, B.L., Urie, J.S., Atkins, J. W., Pyman, M.F.S. and
Wolstencroft, I.R. (1985). Anthelmintic resistance in nematode parasites of
goats. Australian Veterinary Journal 62, 224-227.
Bastien, O., Kerboeuf, D., Leimbacher, F., Gevrey, J., Nicolas, J.A. and Hubert, J.
(1989). Resistance to benzimidazole anthelmintics in nematode parasites of
sheep in Martinique. Recueil De Medecine Viterinaire 165, 461-467.
Bater, A.K. (1989). Efficacy of oral milbemycin against naturally acquired heart-
worm infection in dogs. In “Proceedings of Heartworm Symposium ’89” (G.F.
Otto, ed.), pp. 107-108. American Heartworm Society, Washington, DC.
Bauer, C., Ullrich, D., Fiege, N., Konig, D., Luft, W. and Burger, H.-J. (1987).
Benzimidazole resistance in Haemonchus contortzts on a sheep farm in southern
Germany. Deutsche Tierarztliche Wochenschrijt 94, 205-206.
Bauer, C., Fiege, N., Klatte, D., Enninga, J. and Burger, H.-J. (1988). A survey on
anthelmintic resistance of trichostrongyles in Hessian sheep flocks. Berliner und
Miinchener Tierarztliche Wochenschrift 101, 185-189.
Bennett, J.L. and Pax, R.A. (1986). Micromotility meter: an instrument designed to
evaluate the action of drugs on motility of larval and adult nematodes. Para-
sitology 93, 34 1-346.
Benz, G., Roncalli, R.A. and Gross, S.J. (1989). Use of ivermectin in cattle, sheep,
goats, and swine. In “Ivermectin and Abamectin” (W.C. Campbell, ed.), pp.
215-229. Springer-Verlag, New York.
Berger, J. (1975). The resistance of a field strain of Haemonchus contortus to five
benzimidazole anthelmintics in current use. Journal of the South African
Veterinary Association 46, 369-372.
Besier, B. and Wroth, R. (1993). Ivermectin resistance in Ostertagia in Western
Australia. Newsletter of the Australian Society for Parasitology March, 1-2.
Besier, R.B. and Hopkins, D.L. (1988). Anthelmintic dose selection by farmers.
Australian Veterinary Journal 65, 193-194.
Beugnet, F. (1992). Presence of benzimidazole-resistant gastro-intestinal strongyle

strains in sheep and goats of West of Lyon. Revue de Mkdecine Vktkrinaire 143,
529-533.
Beveridge, I., Ellis, N.J.S, Riley, M.J. and Brown, T.H. (1990). Prevalence of
resistance in sheep nematode populations to benzimidazole and levamisole
anthelmintics in the high rainfall areas of South Australia. Australian Veterinary
Journal 67,413415.
Biehl, L.G. (1986). Anthelmintics for swine. Veterinary Clinics of North America:
Food Animal Practice 2, 481487.
Bisset, S.A., McMurtry, L.M., Vlassoff, A. and West, C.J. (1988). Anthelmintic
resistance to two drench families in a dairy goat herd: suggestions for future
control options. New Zealand Veterinary Journal 36, 201-203.
Bjem, H., Roepstorff, A., Nansen, P. and Waller, P.J. (1989). A dose-response
investigation on the level of resistance to pyrantel citrate in nodular worms of
pigs. Veterinary Parasitology 31, 259-267.
Bjem, H., Roepstorff, A., Waller, P.J. and Nansen, P. (1990). Resistance to
levamisole and cross-resistance between pyrantel and levamisole in Oeso-
phagostomum quadrispinulatum and Oesophagostomum dentatum of pigs.
Veterinary Parasitology 37, 21-30.
Bjem, H., Monrad, J., Kassuku, A.A. and Nansen, P. (1991a). Resistance to
benzimidazoles in Haemonchus contortus of sheep in Tanzania. Acta Tropica
48,59-67.
Bjom, H., Monrad, J. and Nansen, P. (1991b). Anthelmintic resistance in nematode
parasites of sheep in Denmark with special emphasis on levamisole resistance in
Ostertagia circumcincta. Acta Veterinaria Scandinavica 32, 145-1 54.
Bjprm, H., Sommer, Chr., Schoughd, H., Henriksen, Sv. Aa. and Nansen, P.
(199 lc). Resistance to benzimidazole anthelmintics in small strongyles
(Cyathostominae) of horses in Denmark. Acta Veterinaria Scandinavica 32,
253-260.
Black, W.J.M. (1964). The development of a preventive routine against Nemato-
dirus disease of lambs. I. Administration of bephenium compounds at 21-day
intervals. British Veterinary Journal 120, 301-3 10.
Blagbum, B.L., Hendrix, C.M., Lindsay, D.S., Vaughan, J.L., Mysinger, R.H. and
Hepler, D.I. (1989). Milbemycin: efficacy and toxicity in beagle and collie dogs.
. In “Proceedings of Heartworm Symposium ’89” (G.F. Otto, ed.), pp. 109-1 13.
American Heartworn Society, Washington, DC.
Blair, L.S. and Klei, T.R. (1986). Nematode infections of domestic animals:
extraintestinal infections. In “Chemotherapy of Parasitic Diseases” (W.C.
, Campbell and R.S. Rew, eds), pp. 307-319. Plenum Press, New York.
Blanchflower, S.E., Banks, R.M., Everett, J.R., Manger, B.R. and Reading, C.
(1991). New paraherquamide antibiotics with anthelmintic activity. Journal of
Antibiotics 44,492497.
Blizzard, T.A., Marino, G., Mrozik, H., Fisher, M.H., Hoogsteen, K. and Springer,
J.P. (1989). Chemical modification of paraherquamide. I. Unusual reactions and
absolute stereochemistry. Journal of Organic Chemistry 54, 2657-2663.
Boersema, J.H. (1983a). Possibilities and limitations in the detection of anthel-
mintic resistance. In “Facts and Reflections IV. Resistance of Parasites to
Anthelmintics” (F.H.M. Borgsteede, Sv. Aa. Henriksen and J.H. Over, eds),
’ pp. 207-216. Central Veterinary Institute, Lelystad.
Boersema, J.H. (1983b). Resistentie van nematoden tegen anthelmintica. Tijdschrift
voor Diergeneeskunde 108, 426-429.
Boersema, J.H. (1985a). Chemotherapy of gastrointestinal nematodiasis in rumi-

nants. In “Chemotherapy of Gastrointestinal Helminths” (H. Vanden Bossche,
D. Thienpont and P.G. Janssens, eds), pp. 4 0 7 4 2 . Springer-Verlag, Berlin.
Boersema, J.H. (1985b). Chemotherapy of gastrointestinal nematodiasis in birds.
In “Chemotherapy of Gastrointestinal Helminths” (H. Vanden Bossche, D.
Thienpont and P.G. Janssens, eds), pp. 505-5 19. Springer-Verlag, Berlin.
Boersema, J.H., Lewing-Van Der Wiel, P.J. and Borgsteede, F.H.M. (1982).
Benzimidazole resistance in a field strain of Haemonchus contortus in The
Netherlands. Veterinary Record 110, 203-204.
Boersema, J.H., Borgsteede, F.H.M., Eysker, M., Hendrikx, W.M.L., Jansen, J.
and Smith-Buys, C.M.C. (1987). Prevalence of benzimidazole resistance of
nematodes in sheep in The Netherlands. Research in Veterinary Science 43,
18-21.
Bogan, J., Benoit, E. and Delatour, P. (1987). Pharmacokinetics of oxfendazole in
goats: a comparison with sheep. Journal of Veterinary Pharmacology and
Therapeutics 10, 305-309.
Borgsteede, F.H.M. (1986). Resistance of Cooperia curticei against fenbendazole.
Research in Veterinary Science 41, 423424.
Borgsteede, F.H.M. (1990). Anthelmintic resistance in gastrointestinal nematodes
of herbivorous animals in Europe. In “Resistance of Parasites to Antiparasitic
Drugs” (J.C. Boray, P.J. Martin and R.T. Roush, eds), pp. 81-87. MSD Agvet,
Rahway.
Borgsteede, F.H.M. and Duyn, S.P.J. (1989). Lack of reversion of a benzimidazole
resistant strain of Haemonchus contortus after six years of levamisole usage.
Research in Veterinary Science 47, 270-272.
Borgsteede, F.H.M., Schavemaker, S., Van der Burg, W.P.J., Gaasenbeek, C.P.H.
and Pekelder, J.J. (1991). Increase of anthelmintic resistance in sheep in The
Netherlands. Veterinary Record 129, 430-43 1.
Bottjer, K.P. and Bone, L.W. (1985). Trichostrongylus colubriformis: effect of
anthelmintics on ingestion and oviposition. International Journal for Parasitol-
ogy 15, 501-503.
Bowman, D.D., Lin, D., Johnson, R.C. and Hepler, D.I. (1991a). Effect of milbe-
mycin oxime on adult Ancylostoma caninum and Uncinaria stenocephala in dogs
with experimentally induced infections. American Journal of Veterinary
Research 52, 64-67.
Bowman, J.W., Lee, B.L., Whaley, H.A. and Thompson, D.P. (1991b). Effects of
dihydroavermectin B1, and analogs on stretcher muscle of the lined shore
crab, Pachygrapsus crassipes. Comparative Biochemistry and Physiology 99C,
333-340.
Britt, D.P. (1982). Benzimidazole-resistant nematodes in Britain. Veterinary
Record 110, 343-344.
Britt, D.P. (1986). Levamisole-resistant nematodes. Veterinary Record 118, 8 1.
Britt, D.P. and Clarkson, M.J. (1988). Experimental chemotherapy in horses
infected with benzimidazole-resistant small strongyles. Veterinary Record 123,
2 19-22 1.
Burger, H.-J. and Bauer, C. (1987). Efficacy of four anthelmintics against
benzimidazole-resistant cyathostomes of horses. Veterinary Record 120,
293-296.
Cabaret, J. (1991). Efficacy of netobimin against Muellerius capillaris and resistant
strain of digestive tract strongyles in dairy goats. American Journal of Veterinary
Research 52, 1313-1315.
Calcott, P.H. and Fatig, R.O., 111. (1984). Inhibition of chitin metabolism by
avermectin in susceptible organisms. Journal of Antibiotics 37, 253-259.
Campbell, W.C. (1986). The chemotherapy of parasitic infections. Journal of
Parasitology 72, 45-6 1.
Campbell, W.C. (1989) “Ivermectin and Abamectin.” Springer-Verlag, New York.
Campbell, W.C. (1990). Benzimidazoles: veterinary uses. Parasitology Today 6,
13Ck133.
Campbell, W.C. and Benz, G.W. (1984). Ivermectin: a review of efficacy and
safety. Journal of Veterinary Pharmacology and Therapeutics 7, 1-16.
Campbell, W.C., Fisher, M.H., Stapley, E.O., Albers-Schonberg, G. and Jacob,
T.A. (1983). Ivermectin: a potent new antiparasitic agent. Science 221,823-828.
Campos Pereira, M., Campos, R., Foz R.P.P., Lima, S.B. and Vieira-Bressan,
M.C.R. (1989). Estudo comparativo da eficiCncia de ivermectina, de fenbenda-
zole, de mebendazole e de mebendazole associado a0 citrato de piperazina, no
’ controle de ciatostomineos de equinos da raga Mangalarga Paulista. Revista
Faculdade de Medicina Veterinaria Zootecnia Universidade de Sao Paulo 26,
53-60.
Campos Pereira, M., Kohek, I., Jr, Campos, R., Lima, S.B. and Foz, R.P.P. (1991).
A field evaluation of anthelmintics for control of cyathostomes of horses in
Brazil. Veterinary Parasitology 38, 121-129.
Carmichael, I., Visser, R., Schneider, D. and Soll, M. (1987). Haemonchus
contortus resistance to ivermectin. Journal of the South African Veterinary
Association 58, 93.
Carter, G.T. (1989). New anthelmintic and growth promoting agents from actino-
mycetes. In “Novel Microbial Products for Medicine and Agriculture” (A.L.
Demain, G.A. Somkuti, J.C. Hunter-Cevera and H.W. Rossmore, eds), pp.
229-237. Elsevier, Cambridge.
Carter, G.T., Nietsche, J.A., Goodman, J.J., Torrey, M.J., Dunne, T.S., Borders,
D.B. and Testa, R.T. (1987). LL-F42248a, a novel chlorinated pyrrole antibiotic.
Journal of Antibiotics 40, 233-236.
Cawthome, R.J.G. and Cheong, F.H. (1984). Prevalence of anthelmintic resistant
nematodes in sheep in south-east England. Veterinary Record 114, 562-564.
Cawthome, R.J.G. and Whitehead, J.D. (1983). Isolation of benzimidazole resistant
strains of Ostertagia circumcincta from British sheep. Veterinary Record 112,
274-277.
Chalmers, K. (1985). Detection of benzimidazole resistant Nematodirus spathiger.
New Zealand Veterinary Journal 33, 53.
Chapman, H.D. (1990). Drug resistance in coccidia of the fowl. In “Resistance of
Parasites to Antiparasitic Drugs” (J.C. Boray, P.J. Martin and R.T. Roush, eds),
pp. 33-41. MSD Agvet, Rahway.
Chapman, M.R., Klei, T.R. and French, D.D. (1991). Identification of thiabendazole-
resistant cyathostome species in Louisiana. Veterinary Parasitology 39,293-299.
Charles, T.P., Pompeu, J. and Miranda, D.B. (1989). Efficacy of three broad-
spectrum anthelmintics against gastrointestinal nematode infections of goats.
Veterinary Parasitology 34, 7 1-75.
Cohen, M.L. (1992). Epidemiology of drug resistance: implications for a post-
antimicrobial era. Science 257, 1050-1055.
Coles, G.C. (1986a). Anthelmintics for small ruminants. Veterinary Clinics of
North America: Food Animal Practice 2 , 4 1 1 4 2 1 .
Coles, G.C. (1986b). Anthelmintic resistance to sheep. Veterinary Clinics of North
America: Food Animal Practice 2,423-432.
Coles, G.C. (1988). Strategies for control of anthelmintic-resistant nematodes of
ruminants. Journal of the American Veterinary Medical Association 192,
330-334.
Coles, G.C. (1990a). Recent advances in laboratory models for evaluation of
helminth chemotherapy. British Veterinary Journal 146, 113-1 19.
Coles, G.C. ( 1990b). How widespread are benzimidazole-resistant nematodes of
sheep? Veterinary Record 126, 629.
Coles, G.C. and Simpkin, K.G. (1977). Resistance of nematode eggs to the ovicidal
activity of benzimidazoles. Research in Veterinary Science 22, 386-387.
Coles, G.C., Tritschler, J.P., 11, Giordano, D.J., Laste, N.J. and Schmidt, A.L. (1988).
Larval development test for detection of anthelmintic resistant nematodes.
Cbles, G.C., Giordano, D.J. and Tritschler, J.P., 11. (1989a). Efficacy of levamisole
against immature and mature nematodes in goats with induced infections.
American Journal of Veterinary Research 50, 1074-1075.
Coles, G.C., Folz, S.D. and Tritschler, J.P., 11. (1989b). Motility response of
levamjsole/benzimidazole-resistantHaemonchus contortus larvae. Veterinary
Parasitology 31, 253-257.
Coles, G.C., Hong, C. and Hunt, K.R. (1991). Benzimidazole resistant nematodes
in sheep in southern England. Veterinary Record 128, 44.
Coles, G.C., Bauer, C., Borgsteede, F.H.M., Geerts, S., Klei, T.R., Taylor, M.A.
and Waller, P.J. (1992). World Association for the Advancement of Veterinary
Parasitology (W.A.A.V.P.) methods for the detection of anthelmintic resistance
in nematodes of veterinary importance. Veterinary Parasitology 44,35-44.
Colglazier, M.L., Kates, K.C. and Enzie, F.D. (1969). Anthelmintic activity of
tetramisole, thiabendazole, and purified fine particle phenothiazine against
experimental infections of Haemonchus contortus and Trichostrongylus species
in sheep. Proceedings of the Helminthological Society of Washington 36,68-74.
Colglazier, M.L., Kates, K.C. and Enzie, F.D. (1970). Comparative response of
two ovine isolates of Haemonchus contortus to thiabendazole. Journal of
Conder, G.A., Johnson, S.S., Guimond, P.M., Geary, T.G., Lee, B.L., Winterrowd,
C.A., Lee, B.H. and DiRoma, P.J. (1991). Utility of a Haemonchus contortusljird
(Meriones unguiculatus) model for studying resistance to levamisole. Journal of
Conder, G.A., Zielinski, R.J., Johnson, S.S., Kuo, M.-S.T., Cox, D.L., Marshall,
V.P., Haber, C.L., DiRoma, P.J., Nelson, S.J., Conklin, R.D., Lee, B.L., Geary,
T.G., Rothwell, J.T. and Sangster, N.C. (1992). Anthelmintic activity of
dioxapyrrolomycin. Journal of Antibiotics 45, 977-983.
Conder, G.A., Thompson, D.P. and Johnson, S.S. (1993). Demonstration of co-
resistance of Haemonchus contortus to ivermectin and moxidectin. Veterinary
Record 132, 65 1-652.
Conway, D.P. (1964). Variance in the effectiveness of thiabendazole against
Haemonchus contortus in sheep. American Journal of Veterinary Research 25,
844-845.
Cook, G.C. (1990). Use of benzimidazole chemotherapy in human helminthiasis:
indications and efficacy. Parasitology Today 6, 133-136.
Courtney, C.H. and Sundlof, S.F. (1991). “Veterinary Antiparasitic Drugs”.
Institute of Food and Agricultural Sciences, University of Florida, Gainesville.
Craig, T.M. (1986). Epidemiology and control of gastrointestinal nematodes and
cestodes in small ruminants: Southern United States. Veterinary Clinics of North

America: Food Animal Practice 2, 367-372.
Craig, T.M. and Miller, D.K, (1990). Resistance by Haemonchus contortus to
ivermectin in angora goats. Veterinary Record 126, 580.
Craig, T.M., Hatfield, T.A., Pankavich, J.A. and Wang, G.T. (1992). Efficacy of
moxidectin against an ivermectin-resistant strain of Haemonchus contortus in
sheep. Veterinary Parasitology 41, 329-333.
Curtis, C.F. (1985). Theoretical models of the use of insecticide mixtures for the
management of resistance. Bulletin of Entomological Research 75, 259-265.
da Costa, U.C., Benevenga, S.F. and Santiago, M.A.M. (1985). Nematodirus
spathiger resistance to benzimidazole in sheep in Rio Grande do Sul, Brazil.
Abstracts of the World Association for the Advancement of Veterinary
Parasitology, p. 10.
Daly, J.C. (1990). Resistance management strategies: Theory and practice. In
* “Resistance Management in Parasites of Sheep” (J.A. McKenzie, P.J. Martin
and J.H. Arundel, eds), pp. 72-83. Australian Wool Corporation, Melbourne.
Dash, K.M. (1986a). Multiple anthelmintic resistance in Trichostrongyluscolubri-
formis. Australian Veterinary Journal 63, 45-47.
Dash, K.M. (1986b) Control of helminthosis in lambs by strategic treatment with
closantel and broad-spectrum anthelmintics. Australian Veterinary Journal 63,
4-8.
Denham, D.A. (1990). The wise eye of hindsight. Parasitology Today 6, 136.
Dobson, R.J., Donald, A.D., Waller, P.J. and Snowdon, K.L. (1986). An egg-hatch
assay for resistance to levamisole in trichostrongyloid nematode parasites.
Dobson, R.J., Griffiths, D.A., Donald, A.D. and Waller, P.J. (1987). A genetic
model describing the evolution of levamisole resistance in Trichostrongylus
colubriformis a nematode parasite of sheep. IMA Journal of Mathematics
Applied to Medicine and Biology 4, 279-293.
Dobson, R.J., Barnes, E.H., Birclijin, S.D. and Gill, J.H. (1992). The survival of
Ostertagia circumcincta and Trichostrongylus colubriformis in faecal culture as
a source of bias in apportioning egg counts to worm species. International
Journal for Parasitology 22, 1005-1008.
Dorchies, Ph., Mage, C. and Donat, F. (1991). Study on the activity of 11
anthelmintics for sheep in field conditions. Revue de Midecine Vitirinaire
142, 651-655.
Dorny, P., Claerebout, E., Vercruysse, J., Jalila, A. and Sani, R. (1993). Benzimi-
dazole resistance of Haemonchus contortus in goats in Malaysia. Veterinary
Record 133,423-424.
dos Santos, V.T. and Franco, E.B.O. (1967). A parecimento de Haemonchus
resistene ao radical benzimidazole em Uruguaiana. Proceedings Primer
Congress0 Latino-American0 de Parasitologia, p. 105.
Driscoll, M., Dean, E., Reilly, E., Bergholz, E. and Chalfie, M. (1989). Genetic
and molecular analysis of a Caenorhabditis elegans P-tubulin that conveys
benzimidazole sensitivity. Journal of Cell Biology 109, 2993-3003.
Drudge, J.H., Szanto, J., Wyant, Z.N. and Elam, G. (1964). Field studies on parasite
control in sheep: comparison of thiabendazole, ruelene, and phenothiazine.
American Journal of Veterinary Research 25, 1512-15 18.
Drudge, J.H., Lyons, E.T. and Tolliver, S.C. (1977). Resistance of equine stron-
gyles to thiabendazole: critical tests of two strains. Veterinary Medicinelsmall
Animal Clinician 72, 433-435, 437438.
Drudge, J.H., Lyons, E.T., Swerczek, T.W. and Tolliver, S.C. (1983). Cambenda-
zole for strongyle control in a pony band: selection of a drug-resistant population
of small strongyles and teratologic implications. American Journal of Veterinary
Research 44, 110-1 14.
Drudge, J.H., Tolliver, S.C. and Lyons, E.T. (1984). Benzimidazole resistance of
equine strongyles: critical tests of several classes of compounds against popula-
tion B strongyles from 1977 to 1981. American Journal of Veterinary Research
45, 804-809.
Drudge, J.H., Lyons, E.T., Tolliver, S.C., Lowry, S.R. and Fallon, E.H. (1988).
Piperazine resistance in population-B equine strongyles: a study of selection in
thoroughbreds in Kentucky from 1966 through 1983. American Journal of
Veterinary Research 49, 986-994.
Dydge, J.H., Lyons, E.T., Tolliver, S.C. and Fallon, E.H. (1990). Phenothiazine
in the origin of benzimidazole resistance in population-B equine stronglyes.
Drudge, J.H., Lyons, E.T. and Tolliver, S.C. (1991). Resistance of population-B
equine strongyles to thiabendazole, oxfendazole, and phenothiazine (1981 to
1987). 'American Journal of Veterinary Research 52, 1308-1312.
Duncan, J.L. and Love, S. (1991). Preliminary observations on an alternative stra-
tegy for the control of horse strongyles. Equine Veterinary Journal 23,226-228.
Dutton, C.J., Gibson, S.P., Goudie, A.C., Holdom, K.S., Pacey, M.S., Ruddock,
J.C., Bu'Luck, J.D. and Richards, M.K. (1991). Novel avermectins produced by
mutational biosynthesis. Journal of Antibiotics 44, 357-365.
Diiwel, D. (1991). How reliable are in vitro tests in diagnosis of benzimidazole
resistance in sheep? Revue de MCdecine VitCrinaire 142, 623-63 1.
Duwel, D., Schmid, K. and Bechmann, G. (1987). Benzimidazole-resistant
Haemonchus contortus in sheep in the Federal Republic of Germany. Berliner
und Miinchener Tierarztliche Wochenschrift 99, 120-123.
Eagleson, J.S. and Bowie, J.Y. ( 1986). Oxfendazole resistance in Trichostrongylus
axei in cattle in Australia. Veterinary Record 119, 604.
Eagleson, J.S., Bowie, J.Y. and Dawkins, H.J.S. (1992). Benzimidazole resistance
in Trichostrongylus axei in Australia. Veterinary Record 131, 317-318.
Echevania, F.A.M. and Trindade, G.N.P. (1989). Anthelmintic resistance by
Haemonchus contortus to ivermectin in Brazil: a preliminary report. Veterinary
Record 124, 147-148.
Echevama, F.A.M., Armour, J. and Duncan, J.L. (1991). Efficacy of some anthel-
mintics on an ivermectin-resistant strain of Haemonchus contortus in sheep.
Echevama, F.A.M., Gennari, S.M. and Tait, A. (1992). Isoenzyme analysis of
Haemonchus contortus resistant or susceptible to ivermectin. Veterinary
Echevarria, F.A.M., Armour, J., Borba, M.F. and Duncan, J.L. (1993a). Survival
and development of ivermectin-resistant or susceptible strains of Haemonchus
contortus under field and laboratory conditions. Research in Veterinary Science
54, 133-139.
Echevama, F.A.M., Gettinby, G. and Hazelwood, S. (1993b). Model predictions
for anthelmintic resistance amongst Haemonchus contortus populations in
southern Brazil. Veterinary Parasitology 47, 3 15-325.
Eddi, C., Bianchin, I., Honer, M.R., Muniz, R.A., Caracostantogolo, J. and Do
Nascimento, Y.A. (1993). Efficacy of doramectin against field nematode
infections of cattle in Latin America. Veterinary Parasitology 49, 39-44.
Edwards, J.R., Wroth, R., de Chaneet, G.C., Besier, R.B., Karlsson, J., Morcombe,
P.W., Dalton-Morgan, G. and Roberts, D. (1986). Survey of anthelmintic resis-
tance in Western Australian sheep flocks. I. Prevalence. Australian Veterinary
Journal 63, 135-138.
Eysker, M. and Boersema, J.H. (1992). The efficacy of moxidectin against Dictyo-
caulus viviparus in cattle. Veterinary Quarterly 14, 79-80.
Eysker, M., Jansen, J., Boersema, J.H. and Lewing-Van Der Wiel, P.J. (1983a).
Development of benzimidazole resistance in a Haemonchus contortus strain in
The Netherlands following fenbendazole treatment of ewes at parturition.
Eysker, M., Jansen, J. and Wemmenhove, R. (1983b). Alternative grazing of horses
and sheep as control for gastrointestinal helminthiasis in horses. Veterinary
Eysker, M., Jansen, J. and Mirck, M.H. (1986). Control of strongylosis in horses
by alternate grazing of horses and sheep and some other aspects of the
epidemiology of strongylidae infections. Veterinary Parasitology 19, 103-1 15.
Eysker, M., Boersema, J.H., Kooyman, F.N.J. and Berghen, P. (1988). Possible
resistance of small strongyles from female ponies in The Netherlands against
albendazole. American Journal of Veterinary Research 49, 995-999.
Eysker, M., Boersema, J.H. and Kooyman, F.N.J. (1989). Emergence from
inhibited development of cyathostome larvae in ponies following failure to
remove them by repeated treatments with benzimidazole compounds. Veterinary
Femex, M., Mittelholzer, M.-L., Reber, R., Stiirchler, D. and Bispham, K. (1990).
A drug combination to overcome and prevent development of drug resistance in
malaria- parasites. In “Resistance of Parasites to Antiparasitic Drugs’’ (J.C.
Boray, P.J. Martin and R.T. Roush, eds), pp. 17-24. MSD Agvet, Rahway.
Fisher, M.H. (1986). Chemistry of antinematodal agents. In “Chemotherapy of
Parasitic Diseases” (W.C. Campbell and R.S. Rew, eds), pp. 239-266. Plenum
Press, New York.
Fisher, M.A., Jacobs, D.E., Grimshaw, W.T.R. and Gibbons, L.M. (1992a). Pre-
valence of benzimidazole-resistancein equine cyathostome populations in south
east England. Veterinary Record 130, 3 15-3 18.
Fisher, M.A., Jacobs, D.E. and Jones, P.A. (1992b). Field evaluation of an
. albendazole intraruminal capsule against benzimidazole-resistant Haemonchus
contortus. Veterinary Record 130, 35 1-352.
Folz, S.D., Pax, R.A., Thomas, E.M., Bennett, J.L., Lee, B.L. and Conder, G.A.
( 1987). Motility response of benzimidazole-resistant Haemonchus contortus
, larvae to several anthelmintics. Proceedings of the Helminthological Society
of Washington 54, 249-253.
Geary, T.G., Nulf, S.C., Favreau, M.A., Tang, L., Prichard, R.K., Hatzenbuhler,
N.T., Shea, M.H., Alexander, S.J. and Klein, R.D. (1992a). Three P-tubulin
cDNAs from the parasitic nematode Haemonchus contortus. Molecular and
Biochemical Parasitology 50, 295-306.
Geary, T.G., Klein, R.D., Vanover, L., Bowman, J.W. and Thompson, D.P.
(1992b). The nervous systems of helminths as targets for drugs. Journal of
Geary, T.G., Sims, S.S., Thomas, E.M., Vanover, L., Davis, J.P., Winterrowd,
C.A., Klein, R.D., Ho, N.F.H. and Thompson, D.P. (1993). Haemonchus
contortus: Ivermectin-induced paralysis of the pharynx. Experimental
Geerts, S., Brandt, J., Kumar, V. and Biesemans, L. (1987). Suspected resistance of
Ostertagia ostertagi in cattle to levamisole. Veterinary Parasitology 23, 77-82.
Geerts, S., Guffens, G., Brandt, J., Kumar, V. and Eysker, M. (1988). Benzimidazole
resistance of small strongyles in horses in Belgium. Vlaams Diergeneeskundig
Tijdschrijl57, 20-26.
Geerts, S., Brandt, J., Borgsteede, F.H.M. and Van Loon, H. (1989). Reliability and
reproducibility of the larval paralysis test as an in vitro method for the detection
of anthelmintic resistance of nematodes against levamisole and morantel tartrate.
Geerts, S., Bertels, G., Balis, B., Brandt, J. and Kumar, V. (1990). Benzimida-
zole resistance in nematodes on a dairy goat farm in Belgium. Vlaams
Diergeneeskundig Tijdschrift 59, 90-92.
Georghiou, G.P. (1986). The magnitude of the resistance problem. In “Pesticide
Resistance: Strategies and Tactics for Management” (National Academy of
Sciences, ed.), pp. 14-43. National Academy Press, Washington, DC.
Gettinby, G. (1989). Computational veterinary parasitology with an application to
chemical resistance. Veterinary Parasitology 32, 57-72.
Gettinby, G., Soutar, A., Armour, J. and Evans, P. (1989). Anthelmintic resistance
and the control of ovine ostertagiasis: a drug action model for genetic selection.
International Journal for Parasitology 19, 369-376.
Gettinby, G., Hazelwood, S. and Armour, J. (1990). Computer models applied to
drug resistance in parasites. In “Resistance of Parasites to Antiparasitic Drugs”
(J.C. Boray, P.J. Martin and R.T. Roush, eds), pp. 213-219. MSD Agvet,
Rahway.
Gevrey, J., Robin, B., Guerrero, C., Hugonnet, L. and Taranchon, P. (1984).
EfficacitC de l’ivermectine sur 2 souches de strongyles digestifs d’origine
ovine. Comparison avec le thiabendazole et le fenbendazole. Science Vkterinaire
Mtdkcine Comparke 86, 145-151.
Gibbons, A. (1992). Exploring new strategies to fight drug-resistant microbes.
Science 257, 1036-1038.
Gibson, T.E. (1975). “Veterinary Anthelmintic Medication”, Third Edition.
Commonwealth Agricultural Bureaux, Slough.
Giles, C.J., Urquhart, K.A. and Longstaffe, J.A. (1985). Larval cyathostomiasis
(immature trichonema-induced enteropathy): a report of 15 clinical cases.
Equine Veterinary Journal 17, 169-201.
Gill, J.H., Redwin, J.M., Van Wyk, J.A. and Lacey, E. (1991). Detection of
resistance to ivermectin in Haemonchus contortus. International Journal for
Parasitology 21, 77 1-776.
Gillham, R.J. and Obendorf, D.L. (1985). Therapeutic failure of levamisole in d a q
goats. Australian Veterinary Journal 62, 426-427.
Giordano, D.J., Tritschler, J.P., I1 and Coles, G.C. (1988). Selection of ivermectin-
resistant Trichostrongylus colubriformis in lambs. Veterinary Parasitology 30,
139-148.
Goa, K.L., McTavish, D. and Clissold, S.P. (1991). Ivermectin: a review of
its antifilarial activity, pharmacokinetic properties and clinical efficacy in
onchocerciasis. Drugs 42, 640-658.
Gonzales, J.C., Muniz, R.A., Farias, A., Goncalves, LC.B. and Rew, R.S. (1993).
Therapeutic and persistent efficacy of doramectin against Boophilus microplus in
cattle. Veterinary Parasitology 49, 107-1 19.
Gordnier, P.M., Brezner, J. and Tanenbaum, S.W. (1987). Chitin metabolism: not a
target of avermectin/milbemycin activity in insects. Journal of antibiotics 40,

110-1 12.
Gottschall, D.W., Theodorides, V.J. and Wang, R. (1990). The metabolism of
benzimidazole anthelmintics. Parasitology Today 6, 115, 1 19-124.
Goudie, A.C., Evans, N.A., Gration, K.A.F., Bishop, B.F., Gibson, S.P., Holdom,
K.S., Kaye, B., Wicks, S.R., Lewis, D., Weatherley, A.J., Bruce, C.I., Herbert, A.
and Seymour, D.J. (1993). Doramectin -a potent novel endectocide. Veterinary
Grabley, S., Hammann, P., Thiericke, R., Wink, J., Philipps, S. and Zeeck, A.
(1993). Secondary metabolites by chemical screening. 21. Clonostachydiol, a
novel anthelmintic macrodiolide from the fungus Clonostachys cylindrospora
(strain FH-A 6607). Journal of Antibiotics 46, 343-345.
Green, P.E., Forsyth, B.A., Rowan, K.J. and Payne, G. (1981). The isolation of
*
a field strain of Haemonchus contortus in Queensland showing multiple
anthelmintic resistance. Australian Veterinary Journal 57, 79-84.
Grieve,R.B., Frank, G.R., Stewart, V.A., Parsons, J.C.,Abraham, D., MacWilliams,
P.S. and Hepler, D.I. (1989). Effect of dosage and dose timing on heartworm
(Dirofilaria immitis) chemoprophylaxis with milbemycin. In “Proceedings
of Heartworm Symposium ’89” (G.F. Otto, ed.), pp. 121-124. American
Heartwom Society, Washington, DC.
Gruner, L., Kerboeuf, D., Beaumont, C. and Hubert, J. (1986). Resistance to
benzimidazole of Haemonchus contortus utkalensis in sheep on Martinique.
Veterinary Record 118, 276.
Gundlach, J.L., Sadzikowski, A.B., Tomczuk, K. and Uchacz, S. (1992). Usefulness
of drugs containing moxidectin in the control of parasites in pigs. Medycyna
Weterynaryjna 48,209-2 1 1.
Hall, C.A., Campbell, N.J. and Carroll, S.N. (1979). Resistance to thiabendazole in
a field population of Ostertagia circumcincta from sheep. Australian Veterinary
Journal 55, 229-23 1.
Hall, C.A., Ritchie, L. and McDonell, P.A. (1981a). Investigations for anthelmintic
resistance in gastrointestinal nematodes from goats. Research in Veterinary
Science 31, 116-1 19.
Hall, C.A., Kelly, J.D., Whitlock, H.V., Martin, I.C.A., McDonnell, P.A. and
Gunawan, M. (1981b). Five generations of selection with benzimidazole and
Ron-benzimidazole anthelmintics against benzimidazole resistant strains of
Haemonchus contortus and Ostertagia spp. in sheep. Research in Veterinary
Science 30, 138-142.
Hall, C.A., Kelly, J.D., Martin, I.C.A., Whitlock, H.V., McDonell, P.A. and
Gunawan, M. (1981~).Changes in response of a benzimidazole resistant strain
of Haemonchus contortus from sheep after passing through calves. Research in
Veterinary Science 30, 143-146.
Hall, C.A., Ritchie, L. and Kelly, J.D. (1982). Effect of removing anthelmintic
selection pressure on the benzimidazole resistance status of Haemonchus con-
tortus and Trichostrongylus colubriformis in sheep. Research in Veterinary
Science 33, 54-51.
Helle, 0. ( 1986). Treatment against parasites with thiabendazole and fenbendazole
in young stallions. Development of resistance against fenbendazole in small
strongylides. Norsk Veterinaertidsskrift 98, 623-63 1.
Hendrickx, M.O., Anderson, L., Boulard, C., Smith, D.G. and Weatherley, A.J.
(1993). Efficacy of doramectin against warble fly larvae (Hypoderma bovis).
CHEMOTHERAF'YOF NEMATODE INFECTIONS OF VETERINARY IMPORTANCE 69
Hennessy, D.R., Sangster, N.C., Steel, J.W. and Collins, G.H. (1993). Comparative
pharmacokinetic behaviour of albendazole in sheep and goats. International
Journal for Parasitology 23, 321-325.
Herd, R.P. (1986a). Parasite control in horses: seasonal use of equine anthel-
mintics. Modern Veterinary Practice 67, 895-898.
Herd, R.P. (1986b). Pasture hygiene: a nonchemical approach to equine endo-
parasite control. Modern Veterinary Practice 67, 36-38.
Herd, R.P. (1986~).Epidemiology and control of equine strongylosis at Newmarket.
Equine Veterinary Journal 18, 447452.
Herd, R.P. (1986d). Epidemiology and control of nematodes and cestodes in small
ruminants: Northern United States. Veterinary Clinics of North America: Food
Animal Practice 2, 355-362.
Herd, R.P. (1992). Choosing the optimal equine anthelmintic. Veterinary Medicine
87, 231-232, 234, 236-239.
Herd, R.P., Streitel, R.H., McClure, K.E. and Parker, C.F. (1984). Control of
hypobiotic and benzimidazole-resistant nematodes of sheep. Journal of the
Amepican Veterinary Medical Association 184, 726-730.
Herlich, H., Rew, R.S. and Colglazier, M.L. (1981). Inheritance of cambendazole
resistance in Haemonchus contortus. American Journal of Veterinary Research
42, 1342-1344.
Himonas, C. and Papadopoulos, E. (1994). Anthelmintic resistance in imported
sheep. Veterinary Record 134, 456.
Hong, C., Hunt, K.R., Harris, T.J., Coles, G.C., Grimshaw, W.T.R. and McMullin,
P.F. (1992). A survey of benzimidazole resistant nematodes in sheep in three
counties of southern England. Veterinary Record 131, 5-7.
Hope, J.J. and Kemp, G.K. (1980). Apparent Trichonema resistance to fenbendazole.
New Zealand Veterinary Journal 28, 80-81.
Horton, R.J. (1990). Benzimidazoles in a wormy world. Parasitology Today 6,
106.
Hosking, B.C. and Watson, T.G. (1991). Benzimidazole-resistant nematodes in
cattle. Journal of the New Zealand Institute of Agricultural Science 26, 53.
Hotson, I.K., Campbell, N.J. and Smeal, M.G. (1970). Anthelmintic resistance in
Trichostrongylus colubriformis. Australian Veterinary Journal 46, 356-360.
Hubert, J. and Kerboeuf, D. (1992). A microlarval development assay for the
detection of anthelmintic resistance in sheep nematodes. Veterinary Record
130.44246.
Hubert, J., Kerboeuf, D., Nicolas, J.A., Dubost, G. and Gayaud, C. (1991).
Resistance of small ruminant nematodes to benzimidazole derivative com-
pounds in Limousin (France). Recueil de Mkdecine VktCrinaire 167, 135-140.
Hughes, P.L. (1988). Further field comment on anthelmintic resistance in sheep
nematodes. New Zealand Veterinary Journal 36, 157-158.
Hunt, K.R. and Taylor, M.A. (1989). Use of the egg hatch assay on sheep faecal
samples for the detection of benzimidazole resistant nematodes. Veterinary
Record 125, 153-154.
Hunt, K.R., Hong, C., Coles, G.C., Simpson, V.R. and Neal, C. (1992).
Benzimidazole-resistant Cooperia curticei from Comwall, England. Veterinary
Record 130, 164.
Hunt, K.R., Hong, C., Coles, G.C. and Jones, T.O. (1994). Benzimidazole-resistant
Trichostrongylus colubriformis from goats in central England. Veterinary
Record 134, 4 2 0 4 2 1.
Isaza, R., Courtney, C.H., and Neal, F.C. (1987). Benzimidazole-resistant
Haemonchus contortus in roan antelope (Hippotragus equinus). Journal of Zoo

Animal Medicine 18, 96-97.
Jackson, F. (1993). Anthelmi$ic resistance - the state of play. British Veterinary
Journal 149, 123-1 38.
Jackson, R.A., Townsend, K.G., Pyke, C. and Lance, D.M. (1987). Isolation of
oxfendazole resistant Cooperia oncophora in cattle. New Zealand Veterinary
Journal 35, 187-189.
Jackson, F., Coop, R.L., Jackson, E., Little, S. and Russel, A.J.F. (1991). Anthel-
mintic resistant nematodes in goats. Veterinary Record 129, 39-40.
Jackson, F., Coop, R.L., Jackson, E., Scott, E.W. and Russel, A.J.F. (1992a).
Multiple anthelmintic resistant nematodes in goats. Veterinary Record 130,
210-211.
Jackson, F., Jackson, E. and Coop, R.L. (1992b).:Evidence of multiple anthelmintic
, resistance in a strain of Teladorsagia circumcincta (Ostertagia circumcincta)
isolated from goats in Scotland. Research in Veterinary Science 53, 371-374.
Jackson, F., Jackson, E., Little, S., Coop, R.L. and Russel, A.J.F. (1992~).Pre-
valence of anthelmintic-resistant nematodes in fibre-producing goats in Scotland.
Veterinary Record 131, 282-285.
Johansen, M.V. (1989). An evaluation of techniques used for the detection of
anthelmintic resistance in nematode parasites of domestic livestock. Veterinary
Research Communications 13, 455466.
Jones, R.M., Logan, N.B., Wheatherley, A.J., Little, A.S. and Smothers, C.D.
(1993). Activity of doramectin against nematode endoparasites of cattle.
Jordi, R. (1980). Untersuchungen zur anthelminthika-resistenz von tricho-
strongyliden des schafes. Schweizer Archiv fur Tierheilkunde 122, 679-694.
Kassuku, A.A. and Tibaijuka, B. (1987). Anthelmintic resistance of a Haemonchus
contortus strain in sheep and goats at Sokoine University of Agriculture.
Proceedings of the Fifth Tanzania Veterinary Association Scientifrc Conference
5, 193-216.
Kelly, J.D. and Hall, C.A. (1979). Resistance of animal helminths to anthelmintics.
Advances in Pharmacology and Chemotherapy 16, 89-1 28.
Kelly, J.D., Whitlock, H.V., Thompson, H.G., Hall, C.A., Martin, I.C.A. and
Le Jambre, L.F. (1978). Physiological characteristics of free-living and para-
&tic stages of strains of Haemonchus contortus, susceptible or resistant to
benzimidazole anthelmintics. Research in Veterinary Science 25, 376385.
Kelly, J.D., Sangster, N.C., Porter, C.J., Martin, I.C.A. and Gunawan, M.
(1981a). Use of guinea pigs to assay anthelmintic resistance in ovine isolates
of Trichostrongylus colubriformis. Research in Veterinary Science 30, 131-137.
Kelly, J.D., Webster, J.H., Griffin, D.L., Whitlock, H.V., Martin, I.C.A. and
Gunawan, M. (1981b). Resistance to benzimidazole anthelmintics in equine
strongyles. I. Frequency, geographical distribution and relationship between
occurrence, animal husbandry procedures and anthelmintic usage. Australian
Veterinary Journal 57, 163-171.
Kemp, G.K. and Smith, C.F. (1982). Anthelmintic resistance survey in New
Zealand. New Zealand Veterinary Journal 30, 141-144.
Kennedy, M.J. and Phillips, F.E. (1993). Efficacy of doramectin against eyewoms
(Thelazia spp.) in naturally and experimentally infected cattle. Veterinary
Kerboeuf, D. and Hubert, J. (1990). Comparative efficacy of three groups of
anthelmintics against sheep nematodes resistant to benzimidazoles. Recueil de

Mkdecine Vktkrinaire 166, 77 1-775.
Kerboeuf, D., Beaumont-Schwartz, C., Hubert, J. and Maillon, M. (1988). Resis-
tance to anthelmintics of gastro-intestinal nematodes. Results of a survey in Val
de Loire (France). Recueil de Midecine Vitirinaire 164, 1001-1006.
Kettle, P.R., Vlassoff, A., Ayling, J.M., McMurtry, L.W., Smith, S.J. and Watson,
A.J. (1982). A survey of nematode control measures used by sheep farmers and
of anthelmintic resistance on their farms. Part 2. South Island excluding the
Nelson region. New Zealand Veterinary Journal 30, 79-8 1.
Kettle, P.R., Vlassoff, A., Reid, T.C. and Horton, C.T. (1983). A survey of
nematode control measures used by milking goat farmers and of anthelmintic
resistance on their farms. New Zealand Veterinary Journal 31, 139-143.
King, A.I. MacG., Love, S. and Duncan, J.L. (1990). Field investigation of
anthelmintic resistance of small strongyles in horses. Veterinary Record 127,
232-233.
Kingsbury, P.A. (1965). Relationship between egg counts and worm burdens of
young sheep. Veterinary Record 77,900-901.
Knight, R.A., Morrison, E.G. and McGuire, J.A. (1967). Anthelmintic activity of
thiabendazole in feed compared with phenothiazine orally in Mississippi lambs.
Journal of the American Veterinary Medical Association 151, 1438-1442.
Kwa, M.S.G., Kooyman, F.N.J., Boersema, J.H. and Roos, M.H. (1993). Effect of
selection for benzimidazole resistance in Haemonchus contortus on P-tubulin
isotype 1 and isotype 2 genes. Biochemical and Biophysical Research
Communications 191, 413419.
Kwa, M.S.G., Veenstra, J.G. and Roos, M.H. (1994). Benzimidazole resistance
in Haemonchus contortus is correlated with a conserved mutation at amino
acid 200 in P-tubulin isotype 1. Molecular and Biochemical Parasitology 63,
299-303.
Lacey, E. (1985). The biochemistry of anthelmintic resistance. In “Resistance in
Nematodes to Anthelmintic Drugs’’ (N. Anderson and P.J. Waller, eds), pp.
69-78. Commonwealth Scientific and Industrial Research Organization, Glebe.
Lacey, E. (1990). Mode of action of benzimidazoles. Parasitology Today 6,
112-1 15.
Lacey, E. and Prichard, R.K. (1986). Interactions of benzimidazoles (BZ) with
tubulin from BZ-sensitive and BZ-resistant isolates of Haemonchus contortus.
Molecular and Biochemical Parasitology 19, 171-1 8 1.
Lacey, E. and Snowdon, K.L. (1988). A routine diagnostic assay for the detection
of benzimidazole resistance in parasitic nematodes using tritiated benzimidazole
carbamates. Veterinary Parasitology 27, 309-324.
Lacey, E., Redwin, J.M., Gill, J.H., Demargheriti, V.M. and Waller, P.J. (1990). A
larval development assay for the simultaneous detection of broad spectrum
anthelmintic resistance. In “Resistance of Parasites to Antiparasitic Drugs”
Rahway.
Le Jambre, L.F. (1976). Egg hatch as an in vitro assay of thiabendazole resistance
in nematodes. Veterinary Parasitology 2, 385-391.
Le Jambre, L.F. (1978). Anthelmintic resistance in gastrointestinal nematodes of
sheep. In “The Epidemiology and Control of Gastrointestinal Parasites of Sheep
in Australia” (A.D. Donald, W.H. Southcott and J.K. Dineen, eds), pp. 109-120.
CSIRO, Melbourne.
Le Jambre, L.F. (1979). Effectiveness of anthelmintic treatment against levamisole-

resistant Ostertagia. Australian Veterinary Journal 55, 65-67.
Le Jambre, L.F. (1990). Malecular biology and anthelmintic resistance in parasitic
nematodes. In “Resistance of Parasites to Antiparasitic Drugs” (J.C. Boray, P.J.
Martin and R.T. Roush, eds), pp. 155-164. MSD Agvet, Rahway.
Le Jambre, L.F. (1993a). Ivermectin-resistant Haemonchus contortus in Australia.
Australian Veterinary Journal 70, 357.
Le Jambre, L.F. (1993b). Molecular variation in trichostrongylid nematodes from
sheep and cattle. Acta Tropica 53, 331-343.
Le Jambre, L.F., Crofton, H.D. and Whitlock, J.H. (1970). An assay technique for
nematode egg hatchability. Transactions of the American Microscopical Society
89, 397406.
Le Jambre, L.F., Martin, P.J. and Webb, R.F. (1979a). Thiabendazole resistance in
field populations of Haemonchus contortus. Australian Veterinary Journal 55,
163-1 66.
Le Jambre, L.F., Royal, W.M. and Martin, P.J. (1979b). The inheritance of
thiabendazole resistance in Haemonchus contortus..Parasitology 78, 107-1 19.
Le Jambre, L.F., Martin, P.J. and Jarrett, R.G. (1982). Comparison of changes in
resistance of Haemonchus contortus eggs following withdrawal of thiabendazole
selection. Research in Veterinary Science 32, 3 9 4 3 .
Lewis, J.A., Wu, C.-H., Levine, J.H. and Berg, H. (1980). Levamisole-resistant
mutants of the nematode Caenorhabditis elegans appear to lack pharmacological
acetylcholine receptors. Neuroscience 5, 967-989.
Lewis, J.A., Fleming, J.T., McLafferty, S., Murphy, H. and Wu, C. (1987). The
levamisole receptor, a cholinergic receptor of the nematode Caenorhabditis
elegans. Molecular Pharmacology 31, 185-193.
Lichtenfels, J.R. and Hoberg, E.P. (1993).The systematics of nematodes that cause
ostertagiasis in domestic and wild ruminants in North America: an update and a
key to species. Veterinary Parasitology 46, 33-53.
Lichtenfels, J.R., Pilitt, P.A. and Lancaster, M.B. (1988). Systematics of the
nematodes that cause ostertagiasis in cattle, sheep and goats in North America.
Veterinary Parasitology 27, 3-1 2.
Logan, N.B., Weatherley, A.J., Phillips, F.E., Wilkins, C.P. and Shanks, D.J.
(1993). Spectrum of activity of doramectin against cattle mites and lice.
-Veterinary Parasitology 49, 67-73.
Lonneux, J.F. and Losson, B. (1992). Field efficacy of injectable and pour-
on moxidectin in cattle naturally infested with Psoroptes ovis (Acarina:
Psoroptidae). Veterinary Parasitology 45, 147-152.
Love, S., McKellar, Q.A. and Duncan, J.L. (1989). Benzimidazole resistance in a
herd of horses. Veterinary Record 124, 560-561.
Love, S.C.J., Johns, W.H. and Coverdale, O.R. (1992). Anthelmintic resistance in
sheep nematodes in the New Zealand region of New South Wales. Australian
Lubega, G.W. and Prichard, R.K. (1991). Beta-tubulin and benzimidazole resis-
tance in the sheep nematode Haemonchus contortus. Molecular and Biochemical
Lumsden, G.G., Quan-Taylor, R., Smith, S.M. and Washbrooke, I.M. (1989). Field
efficacy of ivermectin, fenbendazole and pyrantel embonate paste anthelmintics
in horses. Veterinary Record 125, 497499.
Lyons, E.T., Drudge, J.H., Tolliver, S.C. and Granstrom, D.E. (1990). Anthel-
mintic resistance in equids. I n “Resistance of Parasites to Antiparasitic Drugs”
(J.C. Boray, P.J. Martin and R.T. Roush, eds), pp. 67-80. MSD Agvet, Rahway.
Lyons, E.T., Drudge, J.H., Tolliver, S.C. and Stamper, S. (1992a). Controlled tests
on activity of contemporary parasiticides on natural infections of helminths in
lambs, with emphasis on strains of Haemonchus contortus isolated in 1955.
American Journal of Veterinary Research 53, 9 1-96.
Lyons, E.T., Tolliver, S.C., Drudge, J.H., Granstrom, D.E., Collins, S.S. and
Stamper, S. (1992b). Critical and controlled tests of activity of moxidectin
(CL 301,423) against natural infections of internal parasites of equids. Veterinary
Lyons, E.T., Drudge, J.H., Tolliver, S.C. and Stamper, S. (1993). Controlled tests
of activity of several antiparasitic compounds against natural infections of
Haemonchus contortus and other helminths in lambs from a flock established
in 1962. American Journal of Veterinary Research 54,406-410.
Maclean, J.M., Lewis, D. and Holmes, P.H. (1987). The pathogenesis of
benzimidazole-resistant and benzimidazole-susceptible strains of Tricho-
strongylus colubriformis in the Mongolian gerbil (Meriones unguiculatus).
Journal’of Helminthology 61, 179-1 89.
Maingi, N. (199 la). Resistance to thiabendazole, fenbendazole and levamisole
in Haemonchus and Trichostrongylus species in sheep on a Kenyan farm.
veterinary Parasitology 39, 285-29 1.
Maingi, N. (1991b). Variations in LCso in the egg hatch assay for anthelmintic
resistant trichostrongylid nematode parasites in sheep. Bulletin of Animal Health
and Production in Africa 39, 167-172.
Maingi, N., Scott, M.E. and Prichard, R.K. (1990). Effect of selection pressure
for thiabendazole resistance on fitness of Haemonchus contortus in sheep.
Mair, T.S. and Cripps, P.J. (1991). Benzimidazole resistance in equine strongyles:
association with clinical disease. Veterinary Record 128, 613-614.
Malan, F.S., Gruss, B., Roper, N.A., Ashbumer, A.J. and DuPlessis, C.A. (1988).
Resistance of Libyostrongylus douglassi in ostriches to levamisole. Journal of
the South African Veterinary Association 59, 202-203.
Marriner, S. and Armour, J. (1986). Nematode infections of domestic animals:
gastrointestinal infections. In “Chemotherapy of Parasitic Diseases” (W.C.
Campbell and R.S. Rew, eds), pp. 287-305. Plenum Press, New York.
Martin, P.J. (1989). Selection for thiabendazole resistance in Ostertagia spp. by
low efficiency anthelmintic treatment. International Journal for Parasitology 19,
3 17-325.
Martin, P.J. ( 1990). Ecological genetics of anthelmintic resistance. I n “Resistance
of Parasites to Antiparasitic Drugs” (J.C. Boray, P.J. Martin and R.T. Roush,
eds), pp. 129-139. MSD Agvet, Rahway.
Martin, R.J. and Kusel, J.R. (1992). On the distribution of a fluorescent ivermectin
probe (4“5,7 dimethyl-bodipy proprionylivermectin) in Ascaris membranes.
Martin, P.J. and Le Jambre, L.F. (1979). Larval paralysis as an in vitro assay of
levamisole and morantel tartrate resistance in Ostertagia. Veterinary Science
Communications 3, 159-164.
Martin, P.J. and McKenzie, J.A. (1990). Levamisole resistance in Trichostrongylus
colubriformis: a sex-linked recessive character. International Journal for
Martin, R.J. and Pennington, A.J. (1989). A patch-clamp study of effects of
dihydroavermectin on Ascaris muscle. British Journal of Pharmacology 98,

747-756.
Martin, P.J., Anderson, N.,-Jarrett, R.G., Brown, T.H. and Ford, G.E. (1982).
Effects of a preventive and suppressive control scheme an the development of
thiabendazole-resistance in Ostertagia spp. Australian Veterinary Journal 58,
185-190.
Martin, P.J., Anderson, N., Lwin, T., Nelson, G. and Morgan, T.E. (1984). The
association between frequency of thiabendazole treatment and the development
of resistance in field isolates of Ostertagia spp. of sheep. International Journal
for Parasitology 14, 177-181.
Martin, P.J., Anderson, N. and Jarrett, R.G. (1985). Resistance to benzimidazole
anthelmintics in field strains of Ostertagia and Nematodirus in sheep. Australian
Martin, P.J., Anderson, N., Brown, T.H. and Miller, D.W. (1988a). Changes in
’
resistance of Ostertagia spp. to thiabendazole following natural selection or
treatment with levamisole. International Journal for Parasitology 18, 333-340.
Martin, P.J., McKenzie, J.A. and Stone, R.A. (1988b). The inheritance of thia-
bendazole resistance in Trichostrongylus colubriformis. International Journal
Martin, P.J., Anderson, N. and Jarrett, R.G. (1989). Detecting benzimidazole
resistance with faecal egg count reduction tests and in vitro assays. Australian
May, G.S., Gambino, J., Weatherbee, J.A. and Moms, N.R. (1985). Identification
and functional analysis of beta-tubulin genes by site specific integrative
transformation in Aspergillus nidulans. Journal of Cell Biology 101, 7 12-7 19.
McKenna, P.B. (1981). The diagnostic value and interpretation of faecal egg
counts in sheep. New Zealand Veterinary Journal 29, 129-132.
McKenna, P.B. (1989). Multigeneric resistance to benzimidazole anthelmintics in
sheep. New Zealand Veterinary Journal 37, 62-64.
McKenna, P.B. (1990a). The use of benzimidazole-levamisole mixtures for the
control and prevention of anthelmintic resistance in sheep nematodes: an
assessment of their likely effects. New Zealand Veterinary Journal 38, 4 5 4 9 .
McKenna, P.B. (1990b). The detection of resistance to ivermectin by the faecal egg
count reduction test. New Zealand Veterinary Journal 38, 169-170.
McKenna, P.B. (1990~).The detection of anthelmintic resistance by the faecal
egg count reduction test: an examination of some of the factors affecting
performance and interpretation. New Zealand Veterinary Journal 38, 142-147.
McKenna, P.B. (1991). Resistance to benzimidazole anthelmintics in cattle in New
Zealand. New Zealand Veterinary Journal 39, 154-155.
McKenna, P.B. and Hughes, P.L. (1986). Non-confirmation of anthelmintic
resistance in Ostertagia spp. New Zealand Veterinary Journal 34, 126-127.
McKenna, P.B. and Seifert, D.A. (1985). Morantel resistance in Trichostrongylus
colubriformis. New Zealand Veterinary Journal 33, 166-1 67.
McKenna, P.B. and Watson, T.G. (1987). The comparative efficacy of four broad
spectrum anthelmintics against some experimentally induced trichostrongylid
infections in sheep and goats. New Zealand Veterinary Journal 35, 192-195.
McKenna, P.B., Badger, S.B., McKinley, R.L. and Taylor, D.E. (1990). Simulta-
neous resistance to two or more broad-spectrum anthelmintics by gastrointestinal
nematode parasites of sheep and goats. New Zealand Veterinary Journal 38,
114-1 17.
Michel, J.F. ( 1985). Epidemiology and control of gastrointestinal helminths in
domestic animals. In “Chemotherapy of Gastrointestinal Helminths” (H. Vanden

Bossche, D. Thienpont and P.G. Janssens, eds), pp. 67-123. Springer-Verlag,
Berlin,
Middelberg, A. and McKenna, P.B. ( 1983). Oxfendazole resistance in Nematodirus
spathiger. New Zealand Veterinary Journal 31, 65-66.
Miller, D.K. and Craig, T.M. (1988). Resistance of sheep and goat nematodes in
Texas to anthelmintics. Sheep and Goat, Wool and Mohair, 1988, Texas
Agricultural Experiment Station, Research Report, September, pp. 23-25.
Miller, J.E. and Baker, N.F. (1980). Thiabendazole-resistant strains of
Haemonchus and Ostertagia in California lambs. American Journal of
Miller, J.E., Hembry, F.G., Kearney, M.T., Williams, J.C., Stagg, L.C. and Sims,
p. (1987). Efficacy of levamisole and netobimin against Haemonchus contortus
in lambs in Louisiana. American Journal of Veterinary Research 48, 1403-1406.
Miller, J.E., Olson, T.A., Kearney, M.T., Myers, G.H. and Williams, J.C. (1992).
Effect of fenbendazole molasses supplement block treatment on nematode
infection and subsequent weight gain of weanling beef calves. Veterinary
Mirck, M.H. (1985). Chemotherapy of gastrointestinal nematodiasis in equines.
In “Chemotherapy of Gastrointestinal Helminths” (H. Vanden Bossche,
D. Thienpont and P.J. Janssens, eds), pp. 443-462. Springer-Verlag, Berlin.
Mitchell, G.B.B., Jackson, F. and Coop, R.L. (1991). Anthelmintic resistance in
Scotland. Veterinary Record 129, 58.
Morley, F.H.W. and Donald, A.D. (1980). Farm management and systems of
helminth control. Veterinary Parasitology 6, 105-1 34.
Moya-Borja, G.E., Muniz, R.A., Sanavria, A., Goncalves, L.C.B. and Rew, R.S.
(1993a). Therapeutic and persistent efficacy of doramectin against Dermatobia
hominis in cattle. Veterinary Parasitology 49, 85-93.
Moya-Borja, G.E., Oliveira, C.M.B., Muniz, R.A. and Goncalves, L.C.B.
(1993b). Prophylactic and persistent efficacy of doramectin against Cochliomyia
hominivorax in cattle. Veterinary Parasitology 49, 95-105.
Neu, H.C. (1992). The crisis in antibiotic resistance. Science 257, 1064-1073.
Ngomuo, A.J., Kassuku, A.A. and Ruheta, M.R. (1990). Critical controlled test to
evaluate resistance of field strains of Haemonchus contortus to thiophanate.
Nilsson, O., Lindholm, A. and Christensson, D. (1989). A field evaluation of
anthelmintics in horses in Sweden. Veterinary Parasitology 32, 163-17 1.
Njanja, J.C., Wescott, R.B. and Ruvuna, F. (1987). Comparison of ivermectin and
thiabendazole for treatment of naturally occumng nematode infections of goats
in Kenya. Veterinary Parasitology 23, 205-209.
Obendorf, D.L., Parsons, J. and Nicholls, J. (1986). An egg development test for
the evaluation of benzimidazole resistance in Nematodirus spathiger. Australian
Obendorf, D.L., Nicholls, J., Koen, T. and Lacy, E. (1991). Benzimidazole-
resistant Nematodirus sp. in Tasmania. Australian Veterinary Journal 68,72-73.
Ondeyka, J.G., Goegelman, R.T., Schaeffer, J.M., Keleman, L. and Zitano, L.
( 1990). Novel antinematodal and antiparasitic agents from Penicillium
charlesii. I. Fermentation, isolation and biological activity. Journal of Antibiotics
43, 1375-1379.
Onishi, J.C. and Miller, T.W. (1985). The lack of antifungal activity by avermectin
B1,. Journal of Antibiotics 38, 1568-1572.
Oosthuizen, W.J.T. and Erasmus, J.B. (1993). Efficacy of moxidectin against a

strain of Haemonchus contortus resistant to ivermectin, a benzimidazole and a
salicylanilide. Journal of the South African Veterinary Association 64, 9-12.
Orbach, M.J., Porro, E.B. and Yanofsky, C. (1986). Cloning and characterization of
the gene for beta-tubulin from a benomyl-resistant mutant of Neurospora crassa
and its use as a dominant selectable marker. Molecular and Cellular Biology 6,
2452-246 1.
Orpin, P.G. (1991). Anthelmintic resistance. Veterinary Record 129, 475.
Ostlind, D.A., Mickle, W.G., Ewanciw, D.V., Andriuli, F.J., Campbell, W.C.,
Hernandez, S., Mochales, S. and Munguira, E. (1990). Efficacy of paraherqua-
mide against immature Trichostrongylus colubriformis in the gerbil (Meriones
unguiculatus). Research in Veterinary Science 48, 260-261.
Pankavich, J.A., Berger, H. and Simkins, K.L. (1992). Efficacy of moxidectin,
nemadectin and ivermectin against an ivermectin-resistant strain of Haemonchus
contortus in sheep. Veterinary Record 130, 241-243.
Partosoedjono, S., Drudge, J.H., Lyons, E.T. and Knapp, F.W. (1969). Evaluation
of naphthalophos against Oestrus ovis and a thiabendazole-tolerant strain of
Haemonchus contortus in lambs. American Journal of Veterinary Research
30, 81-83.
Peters, W. (1984). Use of drug combinations. In “Anti-Malarial Drugs 11” (W.E.
Peters and W.H. Richards, eds), pp. 499-509. Springer-Verlag, Berlin.
Pinheiro, A.C. and Echevama, F.A.M. (1990). Susceptibilidade de Haemonchus
spp. em bovinos ao tratamiento anti-helmintico com albendazole e oxfendazole.
Pesquisa Veterinaria Brasileira 10, 19-21.
Pomroy, W.E. and Whelan, N.C. (1993). Efficacy of moxidectin against an
ivermectin-resistant strain of Ostertagia circumcincta in young sheep. Veterinary
Record 132, 416.
Pomroy, W.E., Whelan, N., Alexander, A.M., West, D.M., Stafford, K., Adlington,
B.A. and Calder, S.M. (1992). Multiple resistance in goat-derived Ostertagia
and the efficacy of moxidectin and combinations of other anthelmintics. New
Zealand Veterinary Journal 40, 76-78.
Presidente, P.J.A. (1985). Methods for detection of resistance to anthelmintics. In
“Resistance in Nematodes to Anthelmintic Drugs” (N. Anderson and P.J.
Waller, eds), pp. 13-27. Commonwealth Scientific and Industrial Research
. Organization, Glebe.
Prichard, R.K. (1986). Anthelmintics for cattle. Veterinary Clinics of North
America: Food Animal Practice 2, 489-501.
Prichard, R.K. (1990a). Anthelmintic resistance in nematodes: extent, recent
’ understanding and future directions for control and research. International
Journal for Parasitology 20, 5 15-523.
Prichard, R.K. (1990b). Biochemistry of anthelmintic resistance. In “Resistance of
Parasites to Antiparasitic Drugs” (J.C. Boray, P.J. Martin and R.T. Roush, eds),
pp. 141-146. MSD Agvet, Rahway.
Prichard, R.K., Hall, C.A., Kelly, J.D., Martin, I.C.A. and Donald, A.D. (1980).
The problem of anthelmintic resistance in nematodes. Australian Veterinary
Journal 56, 239-25 1.
Raether, W. (1988). Chemotherapy and other control measures of parasitic diseases
in domestic animals and man. In “Parasitology in Focus” (H. Mehlhom, ed.),
pp. 8 19-85 1. Springer-Verlag, Berlin.
Ranjan, S., Trudeau, C., Prichard, R.K., Van Kutzleben, R. and Carrier, D. (1992).
Efficacy of moxidectin against naturally acquired nematode infections in cattle.
Reinecke, R.K., Louren, M. and Petersen, B. (1991). Overberg research projects.
XI. First stage larval reduction test to assess anthelmintic efficacy ante mortem
in sheep. Onderstepoort Journal of Veterinary Research 58, 285-290.
Reinemeyer, C.R., Rohrbach, B.W., Grant, V.M. and Radde, G.L. (1992). A survey
of ovine parasite control practices in Tennessee. Veterinary Parasitology 42,
111-122.
Rew, R.S. and Fetterer, R.H. (1986). Mode of action of antinematodal drugs. In
“Chemotherapy of Parasitic Diseases” (W.C. Campbell and R.S. Rew, eds), pp.
321-337. Plenum Press, New York.
Riffkin, G.G., Callinan, A.P.L., Freemantle, A.M., Westcott, J.M., Napthine, D.V.
and O’Connor, A.J. ( 1984). Anthelmintic resistance and sheep management
practices in south western Victoria. Australian Veterinary Journal 61, 248-25 1.
Rochette, F. (1985a). Chemotherapy of gastrointestinal nematodiasis in pigs.
D. Thienpont and P.G. Janssens, eds), pp. 463485. Springer-Verlag. Berlin.
Rochette, F. (1985b). Chemotherapy of gastrointestinal nematodiasis in carnivores.
D. Thienpont and P.G. Janssens, eds), pp. 487-504. Springer-Verlag, Berlin.
Roepstorff, A., Bjern, H. and Nansen, P. (1987). Resistance of Oesophagostomum
spp. in pigs to pyrantel citrate. Veterinary Parasitology 24, 229-239.
Rolfe, P.F. (1990). Resistance of Haemonchus contortus to broad and narrow
spectrum anthelmintics. In “Resistance of Parasites to Antiparasitic Drugs’’
Rahway.
Rolfe, P.F., Boray, J.C., Fitzgibbon, C., Parsons, G., Kemsley, P. and Sangster, N.
( 1990). Closantel resistance in Haemonchus contortus from sheep. Australian
Veterinary Journal 67, 29-3 1.
Roos, M.H. (1990). The molecular nature of benzimidazole resistance in helminths.
Parasitology Today 6, 125-127.
Roos, M.H., Boersema, J.H., Borgsteede, F.H.M., Cornelissen, J., Taylor, M. and
Ruitenberg, E.J. (1990). Molecular analysis of selection for benzimidazole
resistance in the sheep parasite Haemonchus contortus. Molecular and
Biochemical Parasitology 43, 77-88.
Rothwell, J.T. and Sangster, N.C. (1993). An in vitro assay utilizing parasitic larval
Haemonchus contortus to detect resistance to closantel and other anthelmintics.
Round, M.C., Simpson, D.J., Haselden, C.S., Glendinning, E.S.A. and Baskerville,
R.E. (1 974). Horse strongyles’ tolerance to anthelmintics. Veterinary Record 95,
5 17-5 18.
Roush, R.T. (1990). Genetics and management of insecticide resistance: lessons for
resistance in internal parasites? In “Resistance of Parasites to Antiparasitic
Drugs’’ (J.C. Boray, P.J. Martin and R.T. Roush, eds), pp. 197-211. MSD
Agvet, Rahway.
Roush, R.T. and McKenzie, J.A. (1987). Ecological genetics of insecticide and
acaricide resistance. Annual Review of Entomology 32, 361-380.
Ryan, W.G., Lumsden, G.G., Smith, S.M. and Taylor, M.A. (1987). Benzimidazole
resistance in equine small strongyles. Veterinary Record 121, 497.
Samson, D., Charleston, W.A.G., Pomroy, W.E. and Alexander, A.M. (1992).
Evaluation of moxidectin for the treatment of internal parasites of cattle. New

Sangster, N.C. (1990). Biochemical diagnosis of anthelmintic resistance. In
“Resistance of Parasites to Antiparasitic Drugs’’ (J.C. Boray, P.J. Martin and
R.T. Roush, eds), pp. 171-176. MSD Agvet, Rahway.
Sangster, N.C., Whitlock, H.V., Russ, I.G., Gunawan, M., Griffin, D.L. and Kelly,
J.D. (1979).Trichostrongylus colubriformis and Ostertagia circumcincta resis-
tant to levamisole, morantel tartrate and thiabendazole: occurrence of field
strains. Research in Veterinary Science 27, 106-1 10.
Sangster, N.C., Riley, F.L.and Collins, G.H. (1988).Investigation of the mechanism
of levamisole resistance in trichostrongylid nematodes of sheep. International
Journal for Parasitology 18,813-8 18.
Sangster, N.C., Rickard, J.M., Hennessy, D.R. and Steel, J.W. (1991a).Disposition
of oxfendazole in goats and efficacy compared with sheep. Research in
Sangster, N.C., Davis, C.W. and Collins, G.H. (1991b).Effects of cholinergic
drugs on longitudinal contraction in levamisole-susceptible and -resistant
Haemonchus contortus. International Journal for Parasitology 21, 689-695.
Sangster, N.C., Rickard, J.M., Collins, G.H., Hennessy, D.R. and Steel, J.W.
(1992).Use of a controlled-release albendazole capsule in goats. Australian
Veterinary Journal 69,67-68.
Santiago, M.A.M. and da Costa, U.C. (1979).Resistencia de Haemonchus con-
tortus, Trichostrongylus colubriformis e Ostertagia spp. ao levamisole. Revista
do Centro de Ciincias Rurais Universidade Federal de Santa Maria, Brasil 9,
315-318.
Santiago, M.A.M., da Costa, U.C. and Benevenga, S.F. (1977).Trichostrongylus
colubriformis resistente ao levamisole. Revista do Centro de Ciincias Rurais
Universidade Federal de Santa Maria, Brasil 7 , 421-422.
Santiago, M.A.M., da Costa, U.C. and Benevenga, S.F. (1979). Haemonchus
contortus e Ostertagia circumcincta resistentes ao levamisole. Revista do
Centro de Ciincias Rurais Universidade Federal de Santa Maria, B r a d 9,
101-102.
Sasaki, T., Takagi, M., Yaguchi, T., Miyadoh, S., Okada, T. and Koyama, M.
(1992).A new anthelmintic cyclodepsipeptide, PF1022A.Journal of Antibiotics
45,692-697.
Sawicki, R.M., Devonshire, A.L., Rice, A.D., Moores, G.D., Petzing, S.M. and
Cameron, A. (1978). The detection and distribution of organophosphate and
carbamate insecticide resistant Myzus persicae in Britain in 1976. Pesticide
Science 9, 189-20 1.
Scherrer, A.M., Pomroy, W.E. and Charleston, W.A.G. (1989). A survey of
anthelmintic resistance of ten goat farms in the Manawatu region in 1988.
New Zealand Veterinary Journal 37, 148-149.
Scholl, P.J., Guillot, F.S.and Wang, G.T. (1992).Moxidectin: systemic activity
against common cattle grubs (Hypoderma lineatum) (Diptera: Oestridae) and
trichostrongyle nematodes in cattle. Veterinary Parasitology 41, 203-209.
Scott, J.G. (1989).Cross-resistance to the biological insecticide abamectin in
pyrethroid-resistant house flies. Pesticide Biochemistry and Physiology 34,
27-3 1.
Scott, E.W. and Armour, J. (1991). Effect of development of resistance to benzi-
midazoles, salicylanilides and ivermectin on the pathogenicity and survival of
Haemonchus contortus. Veterinary Record 128,346-349.
Scott, E.W., Bairden, K., Holmes, P.H. and McKellar, Q.A. (1989). Benzimidazole
resistance in nematodes of goats. Veterinary Record 124, 492.
Scott, E.W., McKellar, Q.A., Armour, J., Coop, R.L., Jackson, F. and Mitchell,
G.B.B. (1990). Incidence of anthelmintic resistance in Scotland, UK. In
“Resistance of Parasites to Antiparasitic Drugs” (J.C. Boray, P.J. Martin and
Scott, E.W., Duncan, J.L., McKellar, Q.A., Coop, R.L., Jackson, F. and Mitchell,
G.B.B. (1991a). Benzimidazole resistance in sheep nematodes. Veterinary
Record 128,618-619.
Scott, J.G., Roush, R.T. and Liu, N. (1991b). Selection of high-level abamectin
resistance from field-collected house flies, Musca domestica. Experientia 47,
288-291.
Sheir-Neiss, G., Lai, M.H. and Moms, N.R. (1978). Identification of a gene for
P-tubulin in Aspergillus nidulans. Cell 15, 639-647.
Shelton, M. (1968). An evaluation of some newer anthelmintics. Journal of Animal
Science 27, 1136.
Shoop, W.L. (1993). Ivermectin resistance. Parasitology Today 9, 154-159.
Shoop, W.L., Egerton, J.R., Eary, C.H. and Suhayda, D. (1990). Anthelmintic
activity of paraherquamide in sheep. Journal of Parasitology 76, 349-35 1.
Shoop, W.L., Eary,C.H., Michael, B.F., Haines, H.W. and Seward, R.L. (1991).
Anthelmintic activity of paraherquamide in dogs. Veterinary Parasitology 40,
339-341.
Shoop, W.L., Haines, H.W., Eary, C.H. and Michael, B.F (1992a). Acute toxicity
of paraherquamide and its potential as an anthelmintic. American Journal of
Shoop, W.L., Michael, B.F., Haines, H.W. and Eary,C.H. (1992b). Anthelmintic
activity of paraherquamide in calves. Veterinary Parasitology 43, 259-263.
Shoop, W.L., Egerton, J.R., Seward, R.L., Eary, C.H., Haines, H.W. and Michael,
B.F. (1993a). Anthelmintic activity of milbemycin oxime against adult and
immature Uncinaria stenocephala in dogs. Australian Veterinary Journal 70,
187-1 88.
Shoop, W.L., Haines, H.W., Michael, B.F. and Eary, C.H. (1993b). Mutual
resistance between avermectins and milbemycins: oral activity of ivermectin
and moxidectin against ivermectin-resistant and susceptible nematodes.
Veterinary Record 133, 4 4 5 4 7 .
Simpkin, K.G. and Coles, G.C. (1978). Instability of benzimidazole resistance in
nematode eggs. Research in Veterinary Science 25, 249-250.
Slocombe, J.O.D. and Cote, J.F. (1977). Small strongyles of horses with cross
resistance to benzimidazole anthelmintics and susceptibility to unrelated
compounds. Canadian Veterinary Journal 18, 212-2 17.
Slocombe, J.O.D., Cote, J.F. and McMillan, I. (1989). Effectiveness of oxibenda-
zole against benzimidazole-resistant strongyles in horses. Canadian veterinary
Journal 30, 663-665.
Smeal, M.G., Gough, P.A., Jackson, A.R. and Hotson, I.K. (1968) The occurrence
of strains of Haemonchus contortus resistant to thiabendazole. Australian
Smith, G. (1990). A mathematical model for the evolution of anthelmintic
resistance in a direct life cycle nematode parasite. International Journal of
Parasitology 20, 9 13-92 1.
Smith-Buijs, C.M.C. and Borgsteede, F.H.M. (1986). Effect of cool storage of
faecal samples containing Haemonchus contortus eggs on the results of an in
vitro egg development assay to test anthelmintic resistance. Research in

Stansfield, D.G., and Hepler P.1. (1991). Safety and efficacy of milbemycin oxime
for parasite control. Canine Practice 16, 11-16.
Steams, T. and Botstein, D. (1988). Unlinked noncomplementation: isolation of
new conditional-lethal mutations in each of the tubulin genes of Saccharomyces
cerevisiae. Genetics 119, 249-260.
Stringfellow, F. (1988). An in vitro test for drug resistance in Haemonchus
contortus. Proceedings of the Helminthological Society of Washington 55,
19-23.
Sutherland, I.A. and Lee, D.L. (1990). A larval paralysis assay for the detection of
thiabendazole resistance in trichostrongyles. Parasitology 100, 131-135.
Sutherland, I.A., Lee, D.L. and Lewis, D. (1988). Detection of benzimidazole
resistance in trichostrongylid nematodes. Parasitology Today 4, 22-24.
’ Sutherland, LA., Lee, D.L. and Lewis, D. (1989). Colorimetric assay for the
detection of benzimidazole resistance in trichostrongyles. Research in
Sykes, A.R., McFarlane, R.G. and Familton, A.S. (1992). Parasites, immunity and
anthelmintic resistance. In “Progress in Sheep and Goat Research” (A.W.
Speedy, ed.), pp. 179-191. CAB International, Wallingford.
Tagawa, M., Takiyama, A., Ejima, H. and Kurokawa, K. (1985). Prophylactic
efficacy of milbemycin D against Dirofilaria immitis infection in dogs.
Japanese Journal of Veterinary Science 47, 787-790.
Taylor, M.A. (1990a). Anthelmintic resistance in nematodes in the UK and Ireland.
In “Resistance of Parasites to Antiparasitic Drugs” (J.C. Boray, P.J. Martin and
Taylor, M.A. (1990b). A larval development test for the detection of anthelmintic
resistance in nematodes of sheep. Research in Veterinary Science 49, 198-202.
Taylor, M.A. and Hunt, K.R. (1988). Field observations on the control of ovine
parasitic gastroenteritis in south-east England. Veterinary Record 123, 241-245.
Taylor, M.A and Hunt, K.R. (1989). Anthelmintic drug resistance in the UK.
Taylor, S.M., Mallon, T.R., Blanchflower, W.J., Kennedy, D.G. and Green, W.P.
(1992). Effects of diet on plasma concentrations of oral anthelmintics for cattle
-and sheep. Veterinary Record 130, 264-268.
Terada, M. (1992). Neuropharmacological mechanism of action of PF1022A, an
antinematode anthelmintic with a new structure of cyclic depsipeptide, on
Angiostrongylus cantonensis and isolated frog rectus. Japanese Journal of
Terada, M., Ishih, A., Tungtrongchitr, A., Sano, M. and Shomura, T. (1993).
Effects of PF1022A on developing larvae of Angiostrongylus costaricensis in
mice, with special reference to route, dose and formulation. Japanese Journal of
Theodorides, V.J., Scott, G.C. and Laderman, M. (1970). Strains of Haemonchus
contortus resistant against benzimidazole anthelmintics. American Journal of
Townsend, L.B. and Wise, D.S. (1990). The synthesis and chemistry of certain
anthelmintic benzimidazoles. Parasitology Today 6, 107-1 12.
Turner, M.J. and Schaeffer, J.M. (1989). Mode of action of ivermectin. In
“Ivermectin and Abamectin” (W.C. Campbell, ed.), pp. 73-88. Springer-
Verlag, New York.
Uhlinger, C. and Johnstone, C. (1984). Failure to reestablish benzimidazole

susceptible populations of small strongyles after prolonged treatment with
non-benzimidazole drugs. Equine Veterinary Science 4, 7-9.
Uhlinger, C., Eetrow, J. and Johnstone, C. (1988). A field evaluation of benzimida-
zole and nonbenzimidazole drugs in a herd of dairy goats. Journal of Veterinary
Internal Medicine 2, 113-1 16.
Uhlinger, C., Fleming, S. and Moncol, D. (1992). Survey for drug-resistant
gastrointestinal nematodes in 13 commercial sheep flocks. Journal of the
American Veterinary Medical Association 201, 77-80.
Uppal, R.P., Yadav, C.L., Godara, P. and Rana, Z.S. (1992). Multiple anthelmintic
resistance in a field strain of Haemonchus contortus in goats. Veterinary
Research Communications 16, 195-198.
Van Aken, D., De Bont, J. and Vercruysse, J. (1989). Benzimidazole resistance in a
field population of Haemonchus contortus from goats in Sri Lanka. Small
Ruminant Research 2, 28 1-287.
Vanden Bossche, H. (1985). Pharmacology of anthelmintics. In “Chemotherapy
of Gastrointestinal Helminths” (H. Vanden Bossche, D. Thienpont and P.G.
Janssens, eds), pp. 125-1 8 1. Springer-Verlag, Berlin.
Van Schalkwyk, P.C. (1984). Die invloed van weerstand en toleransie van
helminte op die doeltreffendheid van wurmmiddels by beeste en skape in die
Republiek van Suid-Afrika. Verhandeling, Department Parasitologie, Fakulteit
Veeartsenykunde, Universiteit van Pretoria.
Van Schalkwyk, P.C. and Schroder, J. (1989). Benzimidazole-resistant Ostertagia
circumcincta from angora goats. Journal of the South African Veterinary
Association 60, 76-78.
Van Wyk, J.A. (1987). Weerstandbiedendheid van wurms teen middels by
kleinveeTeorie, en die huidige stand in Suid-Afrika. In “Worm Resistance
Workshop” (J. Schroder, ed.), pp. 1 1-23. Onderstepoort.
Van Wyk, J.A. and Gerber, H.M. (1980). A field strain of Haemonchus contortus
showing slight resistance to rafoxanide. Onderstepoort Journal of Veterinary
Research 47, 137-142.
Van Wyk, J.A. and Malan, F.S. (1988). Resistance of field strains of Haemonchus
contortus to ivermectin, closantel, rafoxanide and the benzimidazoles in South
Africa. Veterinary Record 123, 226-228.
Van Wyk, J.A. and Van Schalkwyk, P.C. (1990). A novel approach to the control
of anthelmintic-resistant Haemonchus contortus in sheep. Veterinary Parasitol-
ogy 35, 61-69.
Van Wyk, J.A. and Van Wijk, E.F. (1992). Resistance of small strongyles in an
equine stud in South Africa to the benzimidazole anthelmintics. Journal of the
South African Veterinary Association 63, 144-147.
Van Wyk, J.A., Gerber, H.M. and Alves, R.M.R. (1982). Slight resistance to
the residual effect of closantel in a field strain of Haemonchus contortus
which showed an increased resistance after one selection in the laboratory.
Onderstepoort Journal of Veterinary Research 49, 257-262.
Van Wyk, J.A., Malan, F.S., Gerber, H.M. and Alves, R.M.R. (1987). Two field
strains of Haemonchus contortus resistant to rafoxanide. Onderstepoort Journal
of Veterinary Research 54, 143-146.
Van Wyk, J.A., Malan, F.S., Gerber, H.M. and Alves, R.M.R. (1989a). The
problem of escalating resistance of Haemonchus contortus to the modem
anthelmintics in South Africa. Onderstepoort Journal of Veterinary Research
56. 4 1 4 9 .
82 GEORGE A. CONOER AN0 WILLIAM C. CAMPBELL
Van Wyk, J.A., Van Schalkwyk, P.C., Gerger, H.M., Visser, E.L., Alves, R.M.R.
and Van Schalkwyk, L. (1989b). South African field strains of Haemonchus
contortus resistant to the levamisole/morantel groups of anthelmintics.
Van Wyk, J.A., Bath, G.F., Gerber, H.M. and Alves, R.M.R. (1990). A field strain
of Trichostrongylus colubriformis resistant to levamisole and morantel in South
Africa. Onderstepoort Journal of Veterinary Research 57, 1 19-122.
Varady, M., Praslicka, J., Corba, J. and Vesely, L. (1993). Multiple anthelmintic
resistance of nematodes in imported goats. Veterinary Record 132, 387-388.
Vercruysse, J., Domy, P. and Meurrens, K. (1989). Benzimidazole resistance of
nematodes in sheep in Belgium. Veterinary Record 125, 602-603.
Vercruysse, J., Domy, P., Hong, C., Harris, T.J., Hammet, N.C., Smith, D.G.
and Weatherley, A.J. (1993). Efficacy of doramectin in the prevention of
gastrointestinai nematode infections in-grazing cattle. Veterinaj Parasitology
49. 51-59.
Vieira, L.S., Berne, M.E.A., Cavalcante, A.C.R. and Costa, C.A.F. (1992).
Haemonchus contortus resistance to ivermectin and netobimin in Brazilian
sheep. Veterinary Parasitology 45, 111-1 16.
Vieira-Bressan, M.C.R., Campos Pereira, M., Cordova, C.C., Suma, R.P. and
Queiroz, A. (1988). Estudo comparativo da eficzicia de ivermectina, de oxiben-
dazole e de febantel no controle de estrongilideos de equinos de raga purosangue
ingles e puro-sangue airabe. Hora Veterinaria 46, 29-33.
Vizard, A.L. and Wallace, R.J. (1987). A simplified faecal egg count reduction test.
Australian Veterinary Journal 64, 109-1 11.
Vlassoff, A. and Kettle, P.R. (1980). Benzimidazole resistance in Haemonchus
contortus. New Zealand Veterinary Journal 28, 23-24.
Wagland, B.M., Jones, W.O., Hribar, L., Bendixsen, T. and Emery, D.L. (1992). A
new simplified assay for larval migration inhibition. International Journal for
Waller, P.J. (1990). Resistance in nematode parasites of livestock to the benzimi-
dazole anthelmintics. Parasitology Today 6, 127-1 29.
Waller, P.J. ( 1993). Control strategies to prevent resistance. Veterinary Parasitology
46, 133-142.
Waller, P.J., Dobson, R.J., Donald, A.D., Griffiths, D.A. and Smith, E.F. (1985).
Selection studies on anthelmintic resistant and susceptible populations of
Trichostrongylus colubriformis of sheep. International Journal for Parasitology
15, 669-676.
Waller, P.J., Dobson, R.J., Obendorf, D.L. and Gillham, R.J. (1986). Resistance
of Trichostrongylus colubriformis to levamisole and morantel: differences in
relation to selection history. Veterinary Parasitology 21, 255-263.
Waller, P.J., Dobson, R.J. and Axelsen, A. (1988). Anthelmintic resistance in the
field: changes in resistance status of parasitic populations in response to anthel-
mintic treatment. Australian Veterinary Journal 65, 376-379.
Waller, P.J., Donald, A.D., Dobson, R.J., Lacey, E., Hennessy, D.R., Allerton, G.R.
and Prichard, R.K. (1989). Changes in anthelmintic resistance status of
Haemonchus contortus and Trichostrongylus colubriformis exposed to different
anthelmintic selection pressures in grazing sheep. International Journal for
Waller, P.J., Dobson, R.J. and Haughey, K.G. (1990a). The effect of combinations
of anthelmintics on parasite populations in sheep. Australian Veterinary Journal
67, 138-140.
Waller, P.J., Nansen, P. and Bjorn, H. (1990b). Anthelmintic resistance and its
impact on the intensive animal production industries of Scandinavia. In
“Resistance of Parasites to Antiparasitic Drugs’’ (J.C. Boray, P.J. Martin and
Watson, T. (1993). Resistance lasts for ever. The New Zealand Farmer April,
38-39.
Watson, T.G. and Hosking, B.C. (1990). Evidence for multiple anthelmintic
resistance in two nematode parasite genera on a Saanen goat dairy. New
Watson, T.G.and Manley, T.R. (1985). Pharmacokinetics of oxfendazole in red
deer (Cervus elaphus). Research in Veterinary Science 38, 231-233.
Watson, T.G., Hosking, B.C. and Mckee, P.F. (1993). Stability of ivermectin
resistance in a field strain of Ostertagia spp. Proceedings of the New Zealand
Society of Animal Production 53, 1-3.
Weatherley, A.J., Hong, C., Harris, T.J., Smith, D.G. and Hammet, N.C. (1993).
Persistent efficacy of doramectin against experimental nematode infections in
calves. Veterinary Parasitology 49, 45-50.
Webb, R.F. and Ottaway, S.J. (1986). The prevalence of anthelmintic resistance in
sheep nematodes on the central tablelands of New South Wales. Australian
Webb, R.F., McCully, C.H., Clarke, F.L., Greentree, P. and Honey, P. (1979). The
incidence of thiabendazole resistance in field populations of Haemonchus
contortus on the Northern Tablelands of New South Wales. Australian Veter-
inary Journal 55,422426.
Webb, J.D., Burg, J.G. and Knapp, F.W. (1991). Moxidectin evaluation against
Solenopotes capillatus (Anoplura: Linognathidae), Bovicola bovis (Mallophaga:
Trichodectidae), and Musca autumnalis (Diptera: Muscidae) on cattle. Journal of
Economic Entomology 84, 1266-1269.
Wescott, R.B. (1986a). Epidemiology and control of nematodes and cestodes in
small ruminants: Western United States. Veterinary Clinics of North America:
Food Animal Practice 2, 363-366.
Wescott, R.B. (1986b). Anthelmintics and drug resistance. Veterinary Clinics of
North America: Equine Practice 2, 367-380.
Wescott, R.B., Jen, L.W., Hellier, L.E., Stenslie, J.L. and Torbeck, R.L. (1982).
Efficacy of combinations of piperazine and fenbendazole against benzimidazole-
resistant small strongyles in horses. Veterinary MedicinelSmall Animal Clinician
77, 247-249.
Wescott, R.B., Jen, L.W., Hellier, L.E., Schaefer, D.C. and Traub, J.L. (1985).
Treating for benzimidazole-resistant small strongyles (cyathostomes). Veterinary
Medicine 80, 60-64.
West, D.M., Pomroy, W.E., Probert, A.D. and Charleston, W.A.G. (1989). Multi-
generic resistance to benzimidazole anthelmintics in four sheep flocks. New
Whitlock, H.V., Kelly, J.D., Porter, C.J., Griffin, D.L. and Martin, I.C.A. (1980). In
vitro field screening for anthelmintic resistance in strongyles of sheep and
horses. Veterinary Parasitology 7, 2 15-232.
Wicks, S.R., Kaye, B., Weatherley, A.J., Lewis, D., Davison, E., Gibson, S.P. and
Smith, D.G. (1993). Effect of formulation on the pharmacokinetics and efficacy
of doramectin. Veterinary Parasitology 49, 17-26.
Williams, J.C. (1991). Efficacy of albendazole, levamisole and fenbendazole
against gastrointestinal nematodes of cattle, with emphasis on inhibited early

fourth stage Ostertagia ostertagi larvae. Veterinary Parasitology 40, 59-7 1.
Williams, J.C.. Corwin,-R.M., Craig, T.M. and Wescott, R.B. (1986). Control
strategies for nematodiasis in cattle. Veterinary Clinics of North America:
Food Animal Practice 2, 247-260.
Williams, J.C., Knox, J.W., Marbury, K.S., Swalley, R.A. and Eddi, C.S. (1991).
Efficacy of levamisole against Ostertagia ostertagi in Louisiana cattle during
maturation of inhibited larvae (September) and during minimal inhibition
(DecembedJanuary). Veterinary Parasitology 40,73-85.
Williams, J.C., Barras, S.A. and Wang, G.T. (1992a). Efficacy of moxidectin
against gastrointestinal nematodes of cattle. Veterinary Record 131, 345-347.
Williams, J.C., Nault, C., Ramsey, R.T. and Wang, G.T. (1992b). Efficacy of
Cydectinm moxidectin 1% injectable against experimental infections of Dictyo-
caulus viviparus and Bunostomum phlebotomum superimposed on natural gas-
trointestinal infections in calves. Veterinary Parasitology 43, 293-299.
Yadav, C.L. (1990). Fenbendazole resistance in Haemonchus contortus of sheep.
Veterinary Record 126,586.
Yadav, C.L. and Uppal, R.P. (1992). Levamisole-resistant Haemonchus contortus
in goats. Veterinary Record 130,228.
Yadav, C.L., Uppal, R.P. and Kalra, S. (1993). An outbreak of haemonchosis
associated with anthelmintic resistance in sheep. International Journal of
Yamazaki, M., Okuyama, E., Kobayashi, M. and Inoue, H. (1981). The structure of
paraherquamide, a toxic metabolite from Penicillium paraherquei. Tetrahedron
Letters 22, 135-136.
Zimmerman, G.L., Hoberg, E.P. and Pankavich, J.A. (1992). Efficacy of orally
administered moxidectin against naturally acquired gastrointestinal nematodes in
cattle. American Journal of Veterinary Research 53, 1409-1410.
Parasites as Indicators of Water Quality and the
Potential Use of Helminth Transmission in
Marine Pollution Studies
K. MacKenzie
SOAFD Marine Laboratory, PO Box 101, Victoria Road, Aberdeen, AB9

8DB. UK
H.H. Williams
Department of Zoology, The National Museum of Wales, Cathays Park,

Cardiff, CFI 3NP and School of Pure and Applied Biology, University of
Wales College of Cardiff, Cardiff, CFl 3TL, UK
B. Williams
Department of Zoology, The National Museum of Wales, Cathays Park,

Cardifl, CFI 3NP, UK
A.H. McVicar
SOAFD Marine Laboratory, PO Box 101, Victoria Road, Aberdeen, AB9

8DB, UK
and
R. Siddall
Department of Biology, University of Jyvaskyla, Seminaarinkatu 15,

SF-40100 Jyvaskyla, Finland
ADVANCES IN PARASITOLOGY VOL 35 Copyright 0 1995 Academic Press Limited

ISBN 0-12-0317354 All rights of reproduction in any form reserved
86 K. MACKENZIE ET AL
1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 86
2. Parasites and Pollution ......................... 87
2.1. Hydrocarbon pollution ....................... 87
2.2. Heavy metals . . . . . . . . . . . . . . . . . . . . . . . . . . . . 92
2.3. Thermal pollution . . . . . . . . . . . . . . . . . . . . . . . . . . 94
2.4. Other forms of pollution ...................... 98
3. Effects of Natural Environmental Factors . . . . . . . . . . . . . . . . . 105
4. Helminth Life Cycles, Transmission Processes and Water Quality ... . 109
5. Guidelines and Procedures for Selecting Hosts and Parasites ..... . 126
5.1. Guidelines ............................. 126
5.2. Procedures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 127
6. Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 128
Acknowledgements . . . . . . . . . . . . . . . . . . . . . . . . . . 129
References ............................... 129
1. INTRODUCTION
The need for sensitive biological indicators to monitor the effects of

pollutants on marine organisms was highlighted by McIntyre and Pearce
(1980) and Thulin (1986). Good indicators must be exceptionally sensitive
to environmental change so that a significant reduction in their numbers
can be used as a warning of deteriorating conditions before the majority of
less-sensitive organisms are seriously affected. The potential value of
parasites in this role was pointed out by one of us (HHW) as far back as
1978 (see Cole, 1979). About one hundred papers directly concerned with
the effects of pollution on fish parasites have been published since 1980,
three of which are major reviews of the subject. This is a complex area of
research because it is difficult to link parasite population levels with
pollution without considering a number of other abiotic and biotic factors
which might be involved. The problem is exacerbated by the need to assess
accurately the degree and nature of pollution. With these difficulties in
mind this review assesses the potential for research on aquatic parasites,
with the emphasis on helminths, as pollution indicators.
Since 1980 attempts to use parasites as indicators of marine pollution
have concentrated on the pathology associated with parasitic infections
(Overstreet and Howse, 1977; Sindermann, 1979; McIntyre and Pearce,
1980; Yevich and Barszcz, 1983; Moller, 1985, 1987a, b; McVicar, 1986;
Overstreet, 1988, 1993). McVicar (1986) pointed to lack of sensitivity as
one of the main limitations of pathology as a monitoring tool. That is, most
obvious pathological conditions are the end result of a sequence of events
that may have occurred over a prolonged period and therefore record
changes in environmental conditions dating back some considerable
time. This is particularly true of pathology resulting from infections with
PARASITES AS INDICATORS OF WATER QUALITY 87
metazoan parasites, which have longer generation times than micro-

organisms such as viruses, bacteria and protozoans, and which con-
sequently take longer to build up their populations to levels likely to
cause disease. We suggest that a more rapid response to environmental
change is likely to be observed by monitoring the transmission processes of
helminth parasites.
There are good reasons for focusing on parasitic organisms in general,
and helminth parasites in particular, in the search for highly sensitive
indicators. First, there are more parasitic than free-living species (Burn,
1980; May, 1988). Second, since helminth parasites have complex life
cycles and the different developmental stages have widely differing
requirements, each stage must be assessed separately for sensitivity to
environmental change, thereby widening the choice of potential indica-
tors. Khan and Thulin (1991) reviewed the effects of pollutants on parasites
of aquatic animals, and suggested areas requiring further study. In this
review our aims are: (1) to update Khan and Thulin (1991); (2) to highlight
the complexity of this area of research because of the need to consider the
effects of natural environmental factors on the ecology of marine parasites;
(3) in planning research, to highlight the potential value of concentrating
on the delicate free-living and often short-lived transmission stages of well-
known helminth life cycles; and (4) to suggest selection criteria for the use
of host-parasite systems in marine pollution monitoring.
2. PARASITES AND POLLUTION
Many of the papers claiming that pollution influences the numbers and
distribution of marine parasites deal with three specific categories of
pollution - hydrocarbon, heavy metal and thermal. We therefore consider
these categories separately below.
2.1. Hydrocarbon Pollution
The toxic effects on marine fish of crude oil and its fractions have been
studied since the early 1970s (Blanton and Robinson, 1973; Anderson et al.,
1974a, b, 1977; Gardner, 1975; Di Michele and Taylor, 1978; McCain et al.,
1978; Payne et al., 1978; Dey et al., 1983; Kiceniuk and Khan, 1983, 1987;
Khan, 1987a, b) and histopathological effects have been studied since the
late 1970s (Hawkes, 1977; Hodgins et al., 1977; Solangi and Overstreet,
1982; Haensly et al., 1982; Khan and Kiceniuk, 1984; Grizzle, 1986).
The effects of oil and its components on fish parasites have been
examined by Haensly et al. (1982), Kiceniuk and Khan (1983), Kiceniuk et

al. (1982), Khan and Kiceniuk (1983, 1988) and Khan (1987a, b, 1990).
Barszcz et al. (1978) reported an increase in the numbers of parasites in
oysters exposed to crude oil.
Laboratory experiments and studies of wild-caught fish suggest that
exposure to oil and its components reduces the prevalence and intensity
of gut parasites (nematodes, Kiceniuk and Khan, 1983; Protozoa and
encysted helminths, Haensly et al., 1982), but may increase infection by
gill parasites (monogeneans, Khan and Kiceniuk, 1988; trichodinid ciliates,
Khan, 1990).
2.1.1. Gill and Skin Parasites

Paperna (1975) reported that mullet may have died from increased mono-
genean infection (Benedenia s ~ . in ) the El Bilaim Lagoon on the eastern
shore of the Gulf of Suez where there is heavy pollution from oil leaking
from submarine oil wells. Kiceniuk and Khan (1983) showed that leeches
were directly affected by oil: the fish leech Johanssonia arctica was
allowed to feed, then exposed for 69 days to a water-accommodated
fraction of Venezuelan crude oil. Digestion was slower in oil-exposed
leeches, and reproduction was poorer, with control leeches producing
three times more eggs than exposed leeches. The possible effects of
hydrocarbons on fish parasites are discussed below under headings accord-
ing to increasing complexity of the life cycles and thus the ecology of the
parasites.
When monogeneans (Gyrodactylus spp.) parasitizing the gills of cod
Gadus morhua were exposed to water-soluble fractions of Venezuelan
crude oil at different concentrations, there was no significant difference
in prevalence and intensity of infection between exposed and control fish
(Khan and Kiceniuk, 1988). However, in a group of 34 cod retained for 16
weeks after the end of the oil exposure period, all individual fish were
infected, compared with only 45% of control fish. Additional infection
occurred during the weeks when cod were exposed to oil-free sea water.
Khan and Kiceniuk (1988) suggested that gill imtation by water-soluble
fractions of oil, resulting in epithelial and mucus cell hyperplasia accom-
panied by excessive mucus secretion and coagulation, might have produced
a habitat conducive to parasite infection and reproduction. The authors
quote Hawkes (1977) as finding that oil exposure was associated with an
increase in gill parasites in fish.
In the period 1990-92, three of the present authors (K. MacKenzie, H.H.
Williams and B. Williams) investigated the occurrence of the monogenean
Diclidophora merlangi on the gills of whiting Merlangius merlangus from
both hydrocarbon polluted and relatively clean areas of the North Sea.
Generally higher prevalences were found than those reported in previous

studies (Smith, 1969; Arme and Halton, 1972; Pilcher et al., 1989),
suggesting that infection of whiting with D . merlangi in the North Sea
has increased over the last three decades. Both prevalence and intensity of
infection with D . merlangi were significantly greater in samples from areas
where the sediments were known to be contaminated with hydrocarbons
than in those from relatively clean areas. These results will be presented in
full in a separate publication, but meanwhile further work is required to
establish if there is a direct link between hydrocarbon pollution and
increased D . merlangi infection in North Sea whiting, and what influence
anumber of other biotic and abiotic factors may have on this host-parasite
relationship.
Khan (1990) reported that the prevalence and intensity of infection by
trichodinid gill ciliates of cod and longhorn sculpin Myoxocephalus octo-
decemspinosus was significantly higher in oil-exposed fish than in controls.
Similarly, in wild-caught Alaskan intertidal sculpin Oligocottus maculosus,
collected from a beach contaminated by an oil spill from the Exxon Valdez
five months earlier, the fish had significantly higher prevalences and
intensities of infection than fish collected from uncontaminated beaches.
Khan (1990) suggested that immunosuppression and changes in the gill
habitat caused by pollutants may account for the increase in parasitism.
Studies by Lehtinen et al. (1984), Dabrowska (1974) and Das and
Shrivastava ( 1984) showed increased trichodinid infection associated
with exposure to pollutants other than oil.
Haensly et al. (1982), however, found no significant difference between
prevalence of gill parasites (predominantly peritrich ciliate protozoans with
some encysted helminths) in plaice Pleuronectes platessa from estuaries
polluted with oil from the Amoco Cadiz crude oil spillage, and plaice from
reference (control) sites. Protozoan parasites of the kidneys of plaice were
similar in prevalence and intensity throughout all the fish collected, and so
appear to be unaffected by oil. A difficulty in interpreting the results of
Haensly et al. (1982) is that migration patterns of plaice in the area were
not well documented so length of exposure of fish to the oil was not known.
2.1.2. Blood Parasites

Kiceniuk et al. (1982) investigated the interactions of trypanosome infec-
tion and crude oil exposure in longhorn sculpins and found that the haemo-
globin concentration of the blood was lower in infected fish exposed
to water-soluble fractions of Venezuelan crude oil than in uninfected
oil-exposed fish and infected fish not exposed to oil. Uninfected, oil-
exposed fish had significantly lower total plasma protein than control fish.
Oil exposure thus appears to affect the haematology of sculpins, and
trypanosome infection potentiates the effect. Most gill monogeneans are

blood feeders and may therefore be affected by hydrocarbon pollution via
the host blood.
Eiras (1987) reported a high prevalence (78.6%) of the blood protozoan
Haemogregarina bigemina in blennies Blennius pholis at Foz do Doura
(west coast of Portugal), an area heavily polluted with hydrocarbons, a
lower prevalence (2.9%) at the least polluted site and intermediate
prevalences (16.8% and 29.4%) at two moderately polluted sites.
Khan (1987a) described the effects of chronic exposure to petroleum
hydrocarbons on winter flounder Pseudopleuronectes americanus and cod
infected with the trypanosome Trypanosoma murmanensis. Oil exposure
. caused increased prevalence of infection, higher parasitaemias and
increased mortality in juvenile winter flounder. In adult winter flounder,
increased prevalence of infection was recorded, but parasitaemias were
low. In sub-adult cod, trypanosome infection alone was related to fewest
mortalities (11% of 36 fish), oil alone to 47% of 19 fish and oil plus
infection to most deaths (68% of 17 fish). In surviving cod from the
experimentally infected group, all oil-exposed fish had continued to be
infected and parasitaemias were high (1.3 ? 0.5 X lo4 parasites per ml of
blood), whereas 66% of fish not exposed to oil had lost their infection and
parasitaemias in the infected fish were too low to be estimated accurately.
In adult cod exposed to oil and then challenged with T. murmanensis, 79%
of 19 cod became infected and parasitaemias were higher (3.0 2 0.8 X lo4
parasites per ml of blood) than in fish not exposed to oil before challenge
(38% of 16 fish, parasitaemia 1.8 2 0.9 X lo4 parasites per ml of blood).
Several workers have related immunosuppression in fish and enhancement
of effects of parasites to the presence of pollutants (Boyce and Behrens
Yamada, 1977; Pascoe and Cram, 1977; Pascoe and Woodworth, 1980;
H-ansen et al., 1982; Sakanari et al., 1984; Wojdani and Alfred, 1984;
Fries, 1986). These results showing persistent infection and high para-
sitaemias in fish exposed to oil suggest immunosuppression in the fish,
and demonstrate the synergistic effect in fish disease of oil (as a stressor)
and parasites.
2.1.3. Gut Parasites

Kiceniuk and Khan (1983) made the point that a number of aromatic or
heterocyclic compounds (which are components of crude oil) anaesthetize
worms and may be used as antinematode drugs. They describe the “old
farmers’ remedy” of administering a jigger of oil to an animal to de-worm
it. They found that winter flounder exposed to newly oiled sediments had
an average number of the gut digenean Steringophorus furciger (= Fello-
distomum furcigerum) of 1.47 compared with 3.95 in flounder exposed to
oiled sediment which had been weathered for a year, and 6.84 in control
fish. Haensly et al. (1982), in their investigations of parasites of plaice after
the Amoco Cadiz oil spill, found that at their last three collection times,
stomach parasites (Protozoa and encysted helminths) were significantly
less frequent at oiled sites than at reference sites.
Khan and Kiceniuk (1983) found that exposure to water-soluble frac-
tions of oil reduced the prevalence and intensity of infection of the
acanthocephalan Echinorhynchus gadi in cod. Prevalence was 100% and
intensity 4.9 worms/fish in controls compared with 7 1 % and 2.0 worms/fish
in cod treated with water-soluble fractions of Hibernian oil for 81 days.
Venezuelan oil appeared to have a weaker effect on parasitism since
differences in prevalence and intensity of infection in exposed and control
fish were significant only after 140 days exposure. Similarly, exposure to
oil reduced the prevalence and intensity of infection of the digenean
Steringophorusfurciger in the gut of winter flounder.
Khan and Kiceniuk (1983) suggested reasons for a reduction in gut
parasites in fish exposed to oil: there could be direct toxicity to the
parasites, or the effects may be indirect. Indirect effects include modifica-
tions of the gut environment so that it becomes inhospitable to the para-
sites, owing to a change in the fish’s physiology and a change in the
composition of bile, which can happen in cod exposed to oil and hydro-
carbons (Dey et al., 1983). If this is so, cod may be an ideal fish for future
work since it has a distinctive tapeworm Abothrium gadi, and it is well
known that tapeworms have requirements for host-specific bile salts
(Williams and Halvorsen, 1971). Although the life cycle of A. gadi has
not been elucidated, it may be a useful model for investigating changes in
water quality.
Some caution is necessary in setting up and interpreting the results of
laboratory experiments with fish carrying gut parasites. Williams (1966)
and Moller (1976) found that starved fish lost their gut parasites. Moller
found that the rate of ejection was particularly high for parasites not
attached to or only loosely attached to the gut wall, or relying on the host’s
gut contents for nutrition. For example, 84.2% of the acanthocephalan
Echinorhynchus gadi was lost from cod over 46 days, and 91.8%, 100%
and 100% of the nematode Hysterothylacium (= Contracaecum) aduncum
(feeders on gut contents), were lost from eelpout Zoarces viviparus, sculpin
Myoxocephalus scorpius and flounder Platichthys jlesus, respectively, over
76 days. The lowest rates of ejection were 38.2% loss of the mucosa-
feeding nematode Cucullanus heterochrous, 13.9% loss of the digenean
Podocotyle atomon and 6.5% loss of the cestode Bothriocephalus scorpii.
Since cestodes and acanthocephalans have no gut and rely for nutrition on
the active transport of amino acids, fatty acids and monosaccharides
through their body surfaces, it is perhaps to be expected that they would
be lost quickly from starving fish. Williams (1966) found that rays Raja
clavata starved for a week were devoid of infection by the cestodes
Echeneibothrium and Echinobothrium.
In contrast to the above results, Khan and Kiceniuk (1983) recorded the
same prevalence (94%) of the digenean Steringophorus furciger in control
flounder kept unfed for 34 days in aquaria as in freshly captured flounder
necropsied immediately, and the prevalence of the acanthocephalan
Echinorhynchus g a d was 100% in two sets of control fish compared
with 94% in freshly captured cod.
2.2. Heavy Metals
Riggs et al. (1987) reported direct effects of selenium pollution on the

cestode Bothriocephalus acheilognathi in fish at Belews Lake, a cooling
reservoir in North Carolina, USA. At the selenium-polluted site, cestode
fecundity was reduced: the ratio of gravid to non-gravid proglottids and the
number of eggs shed per gravid proglottid were significantly lower than at
the non-polluted site, and there were statistically significant associations
between selenium concentration in tapeworm tissue and fecundity. An
important finding was that cestode reproductive tissue accumulated almost
three times as much selenium as host reproductive tissue. Seasonal patterns
of growth differed according to site. At the non-polluted site cestodes in
fathead minnows Pimephales promelas underwent considerable growth in
autumn as well as in spring and summer, but at the polluted site cestodes
grew primarily in spring and summer. The mean infrapopulation biomass
of cestodes in fathead minnows and mosquito fish Gambusia afJinis at the
polluted site was significantly smaller than at the non-polluted site.
Biotic and abiotic factors may act in a complex way on transmission
processes to give rise to apparently contradictory findings. For instance,
Riggs and Esch (1987) found that autumn-winter peaks in mean infra-
population density of B. acheilognathi in mosquito fish and fathead
rpinnows at the unpolluted site were absent, and mean densities of the
cestode were significantly lower. It appears that at the unpolluted site, the
large diverse community of piscivorous fish reduced the abundance of
mosquito fish and fathead minnows, restricting them to the littoral zone
where cover was abundant. In this zone, the copepod intemediate hosts of
the cestode were relatively uncommon. Thus the presence of the predatory
fish reduced the chances of these two species being infected. At the
polluted site, the fish fauna was impoverished owing to direct toxicity
from selenium, so mosquito fish and fathead minnows fed freely in the
limnetic zones where infected copepods were abundant.
Munkittrick and Dixon (1988) found significantly lower prevalence and
intensity of infection of acanthocephalans in the guts of white suckers

Catostomus commersoni in lakes polluted with zinc and copper. The
macroinvertebrate fauna of the lakes was reduced so loss of intermediate
hosts could account for this effect, though direct toxicity to the parasite
cannot be ruled out.
Mohan and Sommerville (1988) provided evidence of a change in the
immune response of fish exposed to sublethal concentrations of cadmium.
This was linked to significantly higher levels of infection by the protozoan
skin parasite Ichthyophthirius multijiliis in carp Cyprinus carpio. In their
experiments carp were immunized against I. multijiliis infection, and a trial
group of carp was exposed to cadmium for 10 days, then challenged with
the infective stages of the parasite. The control group was not exposed to
cadmium. The intensity of infection after challenge was significantly higher
in the cadmium-exposed fish than in the controls. In another set of experi-
ments, c q were~ exposed to both cadmium and infection by the protozoans
for 21-35 days. Control fish were exposed to infection alone. The cadmium-
exposed carp took longer to develop immunity to the parasites. Ewing et al.
(1982) found a weak positive correlation between susceptibility of channel
catfish Ictalurus punctatus to I . multijiliis and their exposure to sublethal
concentrations of copper (0.45-1.6 mg copper 1-, or 0.00045-0.0016 ppm).
Their work suggests that the response of gill tissue to copper exposure was
important in determining susceptibility of fish to the protozoan.
Where pollutants are toxic to parasite larval stages, the efficiency of
transmission processes may be affected. Copper and zinc toxicity to
parasite larval stages has been studied by Evans (1982a, b). Both metals
bind to active sites on enzymes and so may have widespread effects on a
variety of enzyme-mediated metabolic processes.
Evans (1982a) found that concentrations of copper at 10 ppm (0.01 mg
1-') in soft water significantly reduced the numbers of cercariae of the
freshwater digenean Notocotylus attenuatus that successfully encysted.
Concentrations of zinc at 4, 5 and 10 ppm (0.004, 0.005 and 0.01 mg
1-') in hard water had the same effect. In soft water, zinc was slightly
less toxic. Deaths occurred most frequently in the later stages of cyst
formation: it appears that copper and zinc interfere with the extension from
specialized cells of keratin-like granules which dissociate and unfold to
produce the inner layers of the cyst wall. Once the cyst wall is complete, its
thick and chemically complex character provides an effective barrier, since
viability of metacercarial cysts was not significantly reduced, even after six
weeks' exposure to copper and zinc solutions. However, cercarial shedding
was si nificantly reduced at low copper concentrations: at 0.5 ppm (0.0005
-?
mg 1 ) the intermediate snail hosts died within three days, and even at
lower concentrations (0.1 ppm; 0.0001 mg 1-') of copper and zinc both
snail activity and cercarial shedding was reduced.
Evans (1982b) found that copper and zinc at concentrations of 0.1-10.0

mg 1-' (100-10 000 ppm) reduced survival time and infectivity of cercariae
of the freshwater digenean Echinoparyphium recurvatum. Cercarial infec-
tivity was particularly affected: a 15-min exposure of cercariae to 0.5 mg 1-'
(500 ppm) of copper caused significant reductions in their ability to infect
second intermediate molluscan hosts. Copper toxicity was enhanced in
hard water.
Evans (1982a) referred to Forstner and Whittman's (1979) quote of 0.01
and 0.002 ppm as typical concentrations of zinc and copper in water, and to
Brown's (1977) record of a maximum of 2.4 ppm of zinc and 0.8 ppm copper
in rivers contaminated by mine waste water. These higher concentrations of
soluble metal would severely affect cercarial shedding in N. attenuatus, but
the shed cercariae and metacercarial cysts would be little affected. Bryan
and Hummerstone (1973) and Aston et al. (1974) recorded copper at 1 mg
1-' (loo0 ppm) in water polluted by copper mine .effluent and this is within
the range which affects the efficiency of transmission in both N. atrenuatus
and E . recurvatum. Oxygen depletion due to organic pollutants may increase
toxicity of certain metals such as zinc (Guth et al., 1977).
Siddall et al. (1993) reported decreasing prevalence of larval digenean
parasites in the mollusc Buccinum undatum with increasing levels of
sediment contamination by trace metals from a sewage sludge dumping
site in the Firth of Clyde, Scotland. At a relatively clean reference site 20%
of Buccinum were parasitized compared with a prevalence of 2% in
Buccinum on the periphery of the dump and 15% at a site 3 km north of
the dump. Patterns of parasite prevalence were not correlated with several
host-related or natural environmental factors examined. The gradient of
parasite prevalence was considered to be caused primarily by toxic effects
on the miracidia of trace metals in the sludge, reducing parasite transmis-
sion to the mollusc. The benthic invertebrates in the area were not adversely
affected by sewage sludge, in fact an increase in abundance, biomass and
diversity of these organisms was closely related to the organic enrichment
of the sediments.
2.3. Thermal Pollution
Thermal pollution has complex and widespread effects on parasite faunas.

The effects of thermal effluents on parasitism in the large-mouth bass
Micropterus salmoides by the acanthocephalan Neoechinorhynchus cylin-
d r a m were examined by Eure and Esch (1974). Significantly higher
parasite densities were found in fish from heated water during the winter
months, due to greater densities of possible intermediate host species and
larval parasites in the effluent. Hirshfield et al. (1983) thought that
increased numbers of oligochaetes (suspected first intermediate hosts) in

the discharge canal of a power plant in Chesapeake Bay, USA resulted in a
four times greater prevalence of larval nematodes (Eustrongylides sp.) in
mummichogs Fundulus heteroclitus sampled from the canal than in fish
taken from near the water intake. In another study, thermal loading was
associated with changes in the distribution and abundance of two larval
trematodes in mosquito fish (Ah0 et al., 1976). Esch et al. (1976)
suggested that thermal pollution in a South Carolina reservoir might
have caused an epizootic of the ciliate Episfylis sp. on fish. MacKenzie
et ul. (1976) thought that the elevated water temperature at Hunterston
power station was a prerequisite for the development of the micro-
sporidian Glugea stephani in plaice, absent from nearby coastal waters,
but probably transferred with the fish from more southern, warmer waters.
Hoffman and Putz (1964) found that an increase in temperature from 12°C
to 17-24°C caused the disappearance of the monogenean Gyrodactylus
macrochiri from bluegills Lepomis macrochirus.
Moller (1978b) was of the opinion that the effects of artificial currents
were far more important than elevated temperature at the cooling-water
outflow of a power plant at Kiel Bay where increased transport of plankton
resulted in increased zoobenthos, but no increases in helminth infection
were seen in cod examined. However, several studies indicate that
ambient temperatures strongly influenced growth, maturation, fecundity
and seasonal population dynamics of pseudophyllidean cestodes (Liao and
Shih, 1956; Strazhnik and Davydov, 1975; Granath and Esch, 1983a, b;
Walkey and Korting, 1985) and reproduction and life span in monogeneans
(Bychowsky, 1961; Bauer, 1968; Lester and Adams, 1974). Kamiso and
Olson (1986) found that the faster reproductive rate of Gyrodactylus
stellatus on English sole Parophrys vetulus more than compensated for
the shortened life span at elevated temperatures, so that fish kept at the
highest temperature in the laboratory were the most heavily infected.
Water temperature may affect efficiency of parasite transmission: the
motility of Bothriocephalus acheilognathi coracidia is poor below 20°C
and above 30°C (Granath and Esch, 1983a) and cercariae of the digenean
Echinostoma liei are non-infective below 12°C and decreasingly infective
above 30°C (Evans, 1985). High temperatures may influence the resistance
of the fish host: Evans et al. (1965) found that cold water fish produce an
immune response over a temperature range 5-8"C, and this may be
impaired as temperatures increase.
The work of Pojmanska and colleagues (Pojmiinska et al., 1980;
Pojmainska 1984a, b, c, 1985a, b; Dzika, 1987) demonstrates the com-
plexities associated with thermal pollution. They investigated the parasites
of bream Abramis bruma in Lake Goslawskie (affected by long-term
thermal pollution) and in Lake Goplo (natural temperature fluctuation) in
Poland between 1972 and 1974. It was found that the thermally polluted
Lake Goslawskie had two fewer parasite species than Lake Goplo (with 27
species) and the array of parasites present differed: 19 species were common
to both lakes, but eight species occurring in Lake Goplo were absent from
Lake Goslawskie, and six species in that lake were absent from Lake Goplo.
in the thermally polluted lake fewer fish were infected with Myxosporea, the
monogeneans Ductylogyrus fulcatus and D . wunderi, the digenean Sphuero-
stoma brumue and the cestode Curyophyllueus laticeps. In Lake Goslawskie
the parasitic copepod Ergusilus sieboldi matured earlier and had a longer
reproductive period. Changes in the maturation and recruitment of Curyo-
phyllueus luticeps to new fish hosts produced different patterns in the
dynamics of infrapopulation densities in this parasite, as shown by the
change in prevalence and intensity of infection in Lake Goslawskie.
Pojmhska and Dzika (1987) in a report on data collected at Lake
Goslawskie between 1983 and 1984, found that there had been a number
of changes in the ten years since 1974. The parasite community in the lake
had been reduced by two species and five species had disappeared, with
three new species establishing. Four species decreased in frequency and six
others increased, thus changing the dominance structure of the parasite
community. The number of Ergusilus sieboldi had decreased; although the
higher temperatures allowed a longer reproductive period, it is likely that
the poor tolerance by the free-living stages of the higher temperatures
caused an overall decrease to a very low population density. Since most
Ductylogyrus species specific to bream increased it appears that they could
tolerate the higher temperatures.
Some of the trends suggested by the 1972-74 study were confirmed for
parasites with complex life cycles. They were: the earlier peak in density of
some parasite infrapopulations in fish (Curyophyllueus luticeps and the
digenean Tylodelphys cluvutu); earlier maturation of some species (C.
luticeps); prolongation of reproductive period and time of invasion by
young stages (C. luticeps and the digenean Sphuerostoma muius); and
loss of seasonal occurrence in fish (the digeneans Diplostomum sp. and
Ichthyocotylurus sp.) Pojmhska et al. (1980) regarded changes in the
benthos and littoral invertebrates, which act as intermediate hosts to the
parasites, as the main reason for the difference in the parasite communities
between the thermally polluted lakes and Lake Goplo. In 1983-84
Pojmhska and Dzika (1987) found that devastation of the habitats of
invertebrate intermediate hosts and of the nesting and feeding grounds of
the bird final hosts was much more important than thermal pollution in
influencing the parasite fauna. The authors commented that parasitological
studies have limitations in evaluating disturbance to ecosystems from
thermal pollution, but nevetherless the data indicated some stabilization
in Lake Goslawskie. For example, the appearance of Acunthocephalus
anguillae and the increase in Caryophyllaeus laticeps were evidence that

the invertebrate benthos fauna was recovering, and the population
dynamics of some parasite species were stabilizing in a new pattern which
included earlier maturation, longer reproductive period and blurring or loss
of annual seasonality of parasite occurrence in fish.
Hoglund and Thulin (1989) found that the prevalence and relative
density of Paradiplozoon homoion parasitizing roach in an artificial lake,
warmed by cooling water from the Forsmark nuclear power station in
Sweden, were no differrent from control areas. However, there was a
temporal shift in the parasite life cycle: recruitment of the parasites to the
roach hosts started earlier in the year. It appears that increased mortality of
adult parasites owing to intolerance of raised temperatures offsets the
increase in parasite population which should result from earlier recruitment.
The work of Sankurathri and Holmes (1976) further exemplifies the
complexities associated with thermal pollution. They provided a model
of interspecific interactions between the pulmonate mollusc Physa gyrina
and its parasites and commensals, based on observations at Lake Wabamun
in Alberta, Canada. Parts of the lake are heated by thermal effluent from
power stations, and one of these areas was compared with an unheated area
as a control. Temperature was central in modifying the system of interac-
tions, exerting a direct influence not only on the snails, their digenean
parasites and oligochaete commensals, but also on the three major external
factors which act to modify the system - light, amount of vegetation and
population size of definitive hosts (waterfowl). The higher temperatures
exerted an influence primarily through keeping an area of the lake ice-free
during the winter. This allowed more light to get to the vegetation, which
continued to grow, thus providing food and habitat for the snails. The area
of ice-free water attracted and concentrated waterfowl, the definitive hosts
of the digenean parasites, thus favouring an increase in their rate of
transmission to the snails. Although the increased amount of vegetation
favoured an increase in snail numbers, the waterfowl fed on the snails, thus
reducing the intermediate host population and maintaining the transmission
of parasites to the waterfowl throughout the winter (Sankurathri, 1974). In
the unheated parts of the lake, digenean transmission was interrupted by
winter temperatures, but thermal effluents enhanced the suprapopulations
of all parasites and the populations of the intermediate host, P . gyrina. For
instance, prevalence of metacercarial infection in snails in the heated area
was 75.8% compared with 6.5% in the control area. Seasonal variations in
prevalence of total infections with rediae or sporocysts (germinal sacs)
were different in the two areas. In the control area, infections with germinal
sacs increased in prevalence through the summer and peaked in July at
35-50%, whereas in the heated area the summer prevalence was low, and a
peak occurred in winter (2040% prevalence). Four species of larval
digeneans were found in snails in the control area, compared with seven
species in the heated area.
Sankurathri and Holmes (1976) described an interesting interaction
between the commensal oligochaete Chaetogaster limnaei limnaei, living
on the head, foot and in the mantle cavity of P. gyrina, and the larval
digeneans parasitizing this snail. C.1. limnaei actively ingested a number
of cercariae (Echinoparyphium recurvatum, Echinostoma revolutum,
Notocotylus urbanensis, Trichobilharzia cameroni, Apatemon gracilis,
Zchthyocotylurus erraticus and Cotylurus sp.). In experiments using iso-
lated snails and miracidia of E. recurvatum, the number of miracidia
successfully penetrating and developing was inversely proportional to the
number of oligochaetes present. This was also true for cercarial penetration
and development. If snails were grouped together, the protective effect of
the oligochaetes was enhanced. Even if miracidia or cercariae were not
eaten, the oligochaetes obstructed them: dead cercariae were observed in
dishes containing snails with commensal oligochaetes, but not in control
dishes. Wright ( 1956) recorded a similar phenomenon: Turritella snails
from Millport, Scotland, carried on their opercula colonies of the hydroid
Leuckartiara which, together with their free-swimming medusae, fed on
large numbers of cercariae shed by the snails.
C.1. limnaei was sensitive to higher temperatures: in the heated area of
the lake with water temperatures of 24°C or over the oligochaetes were
present on 36.5% of the snails with a mean intensity of 3.6. Conversely, in
the control area, they occurred on 92.7% of snails, with a mean intensity of
10.1 oligochaetes. The lower mean intensity in the thermally polluted area
would favour parasite transmission to snails.
Thulin (1984) suggested that thermal pollution may have been a factor in
changing the distribution and infection intensity of the digenean eye fluke
Diplostomum sp., a freshwater parasite found in coastal waters of the Baltic
Sea. At one site, near the nuclear plant at Oskarshamn, 90% of cod
examined carried these eye flukes, and a quarter of the fish had very
high infection intensities of 50-200 flukes, causing greying or whitening
Qf the lens. Invasion of a marine host (cod) by a freshwater parasite, and
thermal pollution, were factors regarded as perhaps contributing to high
infection levels.
2.4. Other Forms of Pollution
Several studies report on other forms of pollution such as effluent from

bleached kraft pulp mills, sewage, agricultural and industrial waste. Much
of the work is speculative or inconclusive and there is a clear need for
further carefully planned field and laboratory studies. In some cases fish
PARASITES AS INDICATORS OF WATER QUALllY 99
behaviour may be an important factor: Moller (1978b) and Thulin (1987)

pointed out that fish may be attracted to sewage outlets or warm water
outflows from power stations with the result that an increased population
density of fish favours the spread of ectoparasites and infectious diseases.
Fish exposed to pollutants may modify their behaviour and come into
greater or lesser contact with prey items acting as parasite intermediate
hosts. However, some of the reports cited (Table 1) appear to relate some
kinds of pollution to an increase in gill and skin parasites which have a
direct life cycle (e.g. Protozoa and ectoparasitic monogeneans). This may
be related to stress in fish exposed to pollutants and possibly to immuno-
suppression. These increases in parasite prevalence and intensity may
represent an indirect indication of host response to the presence of pollu-
tants. Other reports indicate that changes in parasite prevalence may be
associated with changes in numbers of invertebrate intermediate hosts
exposed,to pollutants, or in vertebrate hosts living near or visiting a water
body (Table 2). There is some evidence that infected intermediate hosts are
more vulnerable to pollutants, so that differential mortality among infected
and uninfected intermediate hosts may reduce transmission of parasites to
fish. For instance, Guth et al. (1977) found that snails Lymnaea sfagnalis
infected with digenetic trematodes were more vulnerable than uninfected
snails to toxicity from zinc at 24 and 75 ppm. Brown and Pascoe (1989)
showed that mortality of freshwater amphipods (Gammuruspulex) exposed
to cadmium was greater for those infected with the acanthocephalan
Pomphorhynchus laevis than for those uninfected. Changes in parasite
prevalence and intensity owing to effects on intermediate hosts represent
another indirect indication of the presence of pollutants. Other observations
on possible effects of pollution on parasites are given in Table 3. Pollutants
may affect the numbers of parasite larval stages carried by invertebrate
intermediate hosts, or cause changes in pathogenicity of invertebrate
pathogens, which may in turn influence the levels of infection of helminths
in fish hosts. Table 4 summarizes some observations relating to this.
However, findings such as those of Yevich and Barszcz (1983) need to
be treated with caution, since electrophoretic work has indicated that
mussels from the west coast of the USA, earlier assumed to be all Mytilus
edulis, were actually two species, Mytilus galloprovincialis and M. trossu-
lus, and it is possible that these sibling species have different responses to
pollutants and parasitic infection (McDonald and Koehn, 1988; Parker,
1991).
In Nova Scotia, Canada, metazoan parasite communities in yellow eels
Anguilla rostrata from a river affected by acid precipitation were compared
by Cone et al. (1993) with communities in eels from a neighbouring river
that had been limed and deacidified. They found that species richness was
greater and there were more multiple infections in the parasite community
Table 1 Reported effects of pollution on gill and skin parasites of fish.

Type of pollution Parasitesfiosts and observations Reference and locality
Eutrophic waters Monogeneans and the parasitic Valtonen et al. (1987)
branchiuran Argulus foliaceus on gills Finland (lakes)
and skin of roach Rutilus rutilus and
perch Perca fluviatilis. Increased
prevalence attributed to lowered
resistance of fish
Eutrophic lakes Digenean' Rhipidocotyle campanula Valtonen and
metacercariae in roach. Exceptionally Taskinen (1988)
high prevalence of infection possibly Finland (lakes)
due to lowered resistance in the fish
Paper and pulp Skin ciliates Trichodina sp., Apiosoma Valtonen and
mill effluent sp. and Ichthiophthirius multifliis in Koskivaara (1987,
roach. Increase in prevalence of 1989)
infection, possibly due to lowered Finland (lakes)
resistance in hosts
Paper and pulp Monogeneans Dactylogyrus spp. on Koskivaara et al.
mill effluent and gills of roach Rutilus rutilus. Intensity (1991)
eutrophic waters of infection significantly higher in Finland (lakes)
polluted lake in 1988, but not in 1986
Pulp mills Monogeneans Paradiplozoon homoion Thulin et al. (1988)
and Dactylogyrus sp. on roach. Baltic Coast
Decrease in prevalence and intensity
with closeness to mills
Pulp mill effluent The ciliate Trichodina sp. on gills of Lehtinen et al. (1984)
Platichthys flesus. Increased infection Baltic (marine)
in exposed fish. Field and laboratory
observations
Industrial waste, The protozoans Trichodina domergulei Das and Shrivastava
. detergents, and Chilonodella cyprini and the ( 1984)
domestic sewage, digenean Dactylogyrus longicirrus on Uttar Pradesh, India
silt from soil gills of Puntius sp. Fish kills (lake)
erosion associated with high levels of
infection
Ammonia, trace Gill monogeneans, Neodiplectanum Skinner ( 1982)
metals and wenningeri on yellowfin mojarra Florida (marine)
pesticides Gerres cinereus; Ancyrocephalus sp.
on grey snapper Lutjanus griseus; A.
parvus on needlefish Strongylura
timucu. Increased infection in
polluted areas
Pollutants from The ciliate Epistylis sp. on skin of Overstreet and Howse
housing, industry fish. Common in bays subject to (1977)
and agriculture less flushing by tides Mississippi Sound,
Gulf of Mexico
(estuarine/marine)
Table 1 Continued
Type of pollution Parasites/hosts and observations Reference and locality
Low level of The myxosporean Myxobolus exiguus Papema and
dissolved oxygen, on gills of mullet Mugil cephalus Overstreet (1981)
possibly caused and M . auratus. Fish kill associated Crimea (marine)
by pollutants with heavy infection
“Municipal Gill trichodinids on gills of several Dabrowska (1974)
pollution” fish species. Increased infection and Poland (rivers)
gill damage in fish from polluted
stations
from the limed river. The difference was particularly marked in those
parasite groups, notably digeneans, with complex life cycles, this being
in large part attributable to the adverse effects of acidification on the
invertebrate intermediate hosts. Differential susceptibility of the parasites
or their intermediate hosts to acidification appeared to be major factors in
structuring the parasite communities. The authors concluded that the
assemblage of metazoan parasites of fish may be useful as environmental
indicators and may also provide information on the dynamics of altered
food webs.
Valtonen and Taskinen (1988) suggested that the increased prevalence of
metacercariae of the digenean Rhipidocotyle campanula in fish from
eutrophic lakes in Finland might be attributable to lowered resistance in
the fish. Snieszko (1974) and Sindermann (1979) reviewed examples of
disease outbreaks associated with pollutants as environmental stressors. In
the light of studies by O’Neill (1981), Zeeman and Brindley (1981),
Kiceniuk et al. (1982), Sakanari et al. (1984), Wojani and Alfred (1984),
Fries (1986), Khan (1987a) and Mohan and Sommerville (1988) it seems
that increased parasitism could reflect increased susceptibility in fish hosts,
since fish exposed to pollutants showed suppressed antibody synthesis,
damage to lymphoid tissue, changes in numbers of leucocytes, reduced
phagocytosis and damage to the mucus layer protecting skin and gills.
Andrews et al. (1966) found that bluegills Lepomis macrochirus exposed
once to concentrations of the insecticide heptachlor at 0.0125-0.500 ppm
became heavily infected with trematode metacercariae which encysted in
the liver, kidneys and heart, whereas unexposed control fish did not.
Some experimental work appears to lend support to the idea that pollu-
tants increase susceptibility of fish to parasites. In experiments with carp
Cyprinus carpio, Vladimirov and Flerov (1975) exposed fish to 1.5 mg
polychloropinene 1-’ water (0.0015 ppm) for 48 h. Only half the fish
survived this exposure. Surviving fish were challenged with parasitic
invasion by I . multijiliis 1, 7, 15, 30 and 60 days after the time of their
Table 2 Possible effects of pollution on fish parasites via invertebrate inter-

mediate hosts and non-piscine vertebrate hosts.
~~
Type of pollution Parasite/host and observations Reference and locality

Pulp mill effluent Digenean Rhipidocotyle illense Valtonen et al. (1987)
metacercariae in roach. Decrease in Finland lakes
prevalence, attributed to decrease in
numbers of intermediate host, the
freshwater mussel Anodonta piscinalis.
Digenean metacercariae Diplostomum
sp. and Tylodelphus sp. in eyes of
perch and roach. Decreased infection,
attributed to loss of intermediate
mollusc hosts
Industrial pollution Protozoans, monogeneans, the Kostarev (1980)
cestodes Proteocephalus, Russia (reservoirs)
Triaenophorus, Ligula, Digrammai the
acanthocephalans Acanthocephalus and
Neoechinorhynchus, the nematodes
Philometra and Rhabdochona, in several
fish species. Decrease in numbers
associated with corresponding decrease
in intermediate host numbers. Digenean
metacercariae Diplostomum sp.,
Posthodiplostomum cuticola and
Cotylurus sp., in several fish species.
Increase in numbers associated with
larger numbers of birds, attracted by
dying fish. The cestode Caryophyllaeus
laticeps in fish. Increased infection
associated with an increase in
oligochaete populations.
Domestic sewage The digenean metacercaria Kostarev (1980)
Opisthorchis felineus, the cestode Russia (reservoirs)
Diphyllobothrium latum in fish.
Increased infection (these helminths
found in man, dogs, cats and other
fish-eating mammals)
A mixture of Monogenean Macrovalvitrematoides Overstreet and Howse
pollutants resulting micropogoni, digenean (1977)
in low levels of Diplomonorchis leistomi, Mississippi (rivers)
dissolved oxygen; acanthocephalan Dollfusentis Overstreet (1988)
river bottom chandleri in Atlantic croaker
composed of (Micropogon undulatus). Slightly lower
black anaerobic prevalence in polluted river, attributed
mud to lower abundance of intermediate
hosts in case of D . chandleri
Table 2 Continued
Type of pollution Parasitehost and observations Reference and locality
Paper and pulp Numbers of helminth parasite species Anikieva (1982)
plant effluent present in 1 1 fish species reduced in Lake in Karelia,
more heavily polluted zones of lake: Russia
41 species in clean zone versus 20
species in most polluted zone.
Monogeneans most affected: 10 species
in clean zone, 1 (Diplozoon paradoxum)
in most-polluted zone. Most differences
were recorded for fish which shoal in
circumscribed areas, e.g. perch Perca
fluviatilis and roach Rutilus rutilus
Urbdindustrial Reduced diversity of species and Bum (1980)
pollution parasite numbers in winter flounder Raritan Bay (polluted
Pseudopleuronectes americanus in area)
heavily polluted area (five parasites Block Island Sound,
missing: E . gadi, Ascarophis sp., (clean area)
Podocotyle atomon, Cryptocotyle E Coast USA
lingua and a digenean of family
Opecoelidae); this area had reduced
benthic fauna so reduction in
intermediate hosts a likely cause (but
direct pollution effects not ruled out)
Urbdindustrial 50% decrease in parasite species of Sulgostowska (1988)
pollution flounder Platichthys flesus since 1930s Gdansk Bay, Poland
from 14 to 7 species. Some species compared 1980s data
lost: cestodes Caryophyllaeus laticeps, with that of
Proteocephalus sp.; acanthocephalans Janiszewska (1939)
Neoechinorhynchus rutili,
Echinorhynchus salmonis and
Corynosoma semerme; digenean
Brachyphallus crenatus. Remaining spp.
tended to increase: cestode
Bothriocephalus scorpii, digenean
metacercariae of Diplostomum sp.,
acanthocephalans E. gadi (2.5% of fish
infected in 1930s, 13.0%infected
1982-83), Pomphorhynchus laevis
(8.5% of fish infected in 1930s, 31.4%
1982-83); nematodes, Thynnascaris
adunca, Cucullanus minutus (latter, from
50% prevalence to 80%, 1982-83).
Changes in parasite community
attributed to loss of certain intermediate
and even final hosts
Table 3 Other observations of possible effects of pollutants on fish parasites.

Type of pollution Parasites/hosts and observations Reference and locality
Sewage and Digenean larvae in 16 fish species. 300(1988)
pesticides Increased prevalence of Clonorchis Korea (river)
sinensis metacercariae accompanied
by lower intensity
Bleached pulp Some acanthocephalans and cestodes Thulin et al. (1988)
mill effluent; in perch Perca jluviatilis, roach Sweden (lakes)
unbleached pulp Rutilus rutilus and ruffe
mill effluent Gymnocephulus cernuus. No increase
or decrease compared with reference
lake
Coal mud, 23 parasite species (protozoans, Rehulka (1983)
petroleum wastes monogeneans, digeneans, an Czechoslovakia (river)
and high-iron acanthocephalan, nematodes, a ,
effluent crustacean) in 15 fish species. The

parasites survived despite pollution;
high infection intensities were found
for the larval nematodes Raphidascaris
acus in liver and intestine wall of
Neomacheilus barbatulus and Phoxinus
phoxinus
Chlorine in Monogenean Gyrodactylus unicopula MacKenzie et al.
cooling water on gills of plaice Pleuronectes (1976)
from power platessa. Absence of parasite perhaps Scotland (estuary)
station due to chlorine treatment
exposure of polychloropinene. Each fish to be challenged was placed singly

for 12 h in a one litre tank containing approximately 2000 ciliates. After 12
h the fish was removed to a larger tank and the extent of invasion by I.
multifiliis estimated on the fifth or seventh day. For each group of fish the
mean number of parasites per fish (abundance) was compared with that for
control fish which had not been exposed to polychloropinene. Suscepti-
bility to the parasite began to increase between one and seven days after the
time of exposure to the toxin: at seven days the mean number of parasites
per experimental fish was twice that in the controls. Susceptibility con-
tinued to increase throughout the subsequent two months: at 60 days after
exposure, the figure in experimental fish was 33 times that in the controls.
Chronic exposure to the toxin increased susceptibility: in fish exposed to
0.2 mg polychloropinene 1-' for 20 days the mean number of parasites per
fish was 46 times greater than that in the controls. In other experiments,
Vladimirov and Flerov (1975) exposed a parental generation of guppies to
phenol and polychloropinene, and subsequently raised their FI and F2
progeny in clean water. The progeny were five or six times more suscep-
Table 4 Possible effects of pollutants on parasites and pathogens of invertebrate

intermediate hosts.
Type of pollution Observation Reference and
locality
PCBs High levels of parasitism in mussels Yevich and Barszcz
exposed to high PCB pollution. East (1983)
coast mussels (particularly along coast West, East and Gulf
of industrialized area from Boston to coasts of USA,
Cape Henlopen) most heavily (Mussel Watch
parasitized by ciliates, nematodes and Programme)
larval trematodes, e.g. Cercaria
milfordensis
Discharge waters Increased infection intensity of gill Sunila (1987)
of iron and steel ciliates in mussels (M.edulis) in Gulf of Finland
factory polluted area. Infection intensity of (Baltic Sea)
trematode stages and nematodes (International
appeared unrelated to water quality Mussel Programme)
Contaminant Larval trematodes in mussels Auffret (1988)
gradients of M . edulis not correlated with pollutant Brittany
copper, PCBs, gradients in experiments or field sites
polycyclic
aromatic hydrocarbons
(PAHs) water-
accommodated
fraction of diesel oil
Experimental Increase in pathogenicity of the Winstead and
exposure to the protozoan Perkinsus marinus in Couch (1988)
carcinogen DENA American oyster Crassostrea virginica. Florida
(n-nitrosodiethyl- Protozoan reproduced at lower than
mine) usual temperatures, became more
invasive, and the oysters’ haemocytic
defence response was atypically low
tible to I . multijiliis infection than control fish. The implications of this

study are that exposure to heavy metals and certain other toxins may
increase the susceptibility of fish to parasites and disease, that a fish’s
immune system may be affected even two months after exposure to the
toxin, and that the first and second generation progeny of parental fish
exposed to toxin could be affected.
3. EFFECTS OF NATURAL ENVIRONMENTAL FACTORS
In the foregoing review of marine parasites and pollution our main aim was
to update Khan and Thulin (1991). However, much of the work published
up to now in this field is inconclusive and largely unconvincing because

there is little evidence to show that the many natural biotic and abiotic
factors that regulate the numbers and distribution of parasites have been
considered together with the effects of the pollutants. The section below
briefly summarizes the effects of these natural factors on parasite ecology.
The fact that many biotic and abiotic factors affect the numbers and
distribution of marine parasites is now well established in a voluminous
literature. With regard to numbers there is much information on preva-
lence, intensity, mean intensity, relative density or abundance, and diver-
sity, as defined by Margolis et al. (1982). Publications on distribution
address three fundamental patterns, namely regular, random and con-
. tagious (also termed “clumped” or “aggregated” ) distributions. Studies
of helminth parasites in particular provide detailed knowledge of the
ecology of Gyrodactylus spp., Diplozoon paradoxum, Caryophyllaeus
laticeps, Bothriocephalus acheilognathi, Pomphorhynchus laevis and
Transversotrema patialense. Research on these species has focused on
parasite birth and death rates, seasonal infection dynamics and the effects
of host age, diet and feeding behaviour. From field and laboratory studies
information has become available on parasites as regulators of host popu-
lations, and on the consequences of competition between parasites. From
the kind of work referred to above it is clear that the factors listed in Table
5 are the most influential biotic and abiotic factors in marine parasite
ecology.
It has become apparent from recent work on parasites and pollution
Table 5 Principal biotic and abiotic factors identified as affecting marine

parasites.
Biotic Abiotic
Host species Temperature
Schooling of fish Season
Age, length and growth rate of host Latitude/longitude
HDst pathology Salinity
Post-spawning migrations of host Oxygen
Host sex and reproductive behaviour Water mineral contents
Host hormone levels Ammonia
Host immunity responses PH
Host life history (including growth, maturation and Light
longevity)
Host dievfeeding behaviour Depth
Availability of infected intermediate hosts as food Water levels
Site of infection Pollution
Intensity, mean intensity and density of infection
Interactions between parasite species
PARASITES AS INDICATORS OF WATER QUALIW 107
that it is not clear how, and to what extent, the factors referred to in
Table 5 individually and in various combinations, affect the numbers and
distribution of parasites under natural conditions. This problem highlights
the complexity of parasite ecology, which can be defined as the total
relationship of a species to its biotic and abiotic environments.
It follows that the relationship between pollution and marine parasite
ecology is also very complex. The problem is exacerbated by the need (1)
to define the kind of pollution that is involved, and (2) to know the form of
parasite life cycle under investigation - what stages are most susceptible
and what host species are involved.
In some instances a parasite may be directly susceptible to the toxic
effects of pollutants, in which case pollution may reduce infection pre-
valence and intensity. On the other hand, if the host is more susceptible
than the parasite to the pollutant, its resistance to infection may be lowered,
leading tb higher prevalence and intensity. Being parasitized may itself
increase the host’s susceptibility to pollution. Fish mortality due to the
combined effects of pollution and parasitic infection has been demon-
strated experimentally (Perevozchenko and Davydov, 1974; Boyce and
Yamada, 1977; Pascoe and Cram, 1977; Pascoe and Woodworth, 1980).
Further complications arise when the life cycle of the parasite is indirect,
since pollution can affect intermediate or alternative hosts.
It is now accepted that a large number of biotic and abiotic factors affect
the prevalence and abundance of parasites and that temperature is among
the most important of these. Esch (1982) divided abiotic factors into two
groups: those which are determined and predictable in character and those
which are not. In nature, temperature is a determinate predictable character,
e.g. according to season, but through the influence of humans in artificially
heating water with thermal effluent from coal-fired and nuclear electricity
power stations, it is often unpredictable. Most indeterminate factors are
artificial and include fire, flood, artificially elevated temperature, radical
temperature change on a short-term basis and nutrient enrichment of water.
This area of research is an important but little understood aspect of
parasites and pollution because it is well known that water temperature
plays a crucial role in the development and activity of free-living and
ectoparasitic stages of fish helminths. Most of the evidence to support this
statement is based on freshwater fish helminths such as Gyrodactylus,
Dactylogyrus, Caryophyllaeus, Proteocephalus, Bunodera, Allocreadium
and Eustrongylides. The monogenean Dactylogyrus solidus on the gills
of carp Cyprinus carpio is sluggish and easily removable at low tempera-
tures (Bauer, 1962). Since temperature will vary according to the season of
the year at various latitudes and longitudes, it is not surprising that much
has been written on the seasonality and zoogeography of parasitic worms
with many references to water temperature as an all-important factor in
explaining seasonal prevalence and intensity of infection. Much of the

literature, however, refers to the difficulty of drawing conclusions on the
precise effect of temperature on the ecology of parasites without also
considering other biotic and abiotic factors. It is beyond the scope of
this review to discuss the literature on temperature, seasonality and bio-
geography. We merely wish to emphasize the need to consider these factors
before drawing conclusions on the effects of the equally complex factor of
pollution. Most of the references on the effects of temperature on fresh-
water fish helminths are cited in Chubb (1977, 1979, 1980, 1982). The
same principles will apply to its effects on marine helminths.
The parasite faunas of euryhaline spec,ies of fish may change con-
* siderably when the fish move into areas of different salinity (Janiszewska,
1939; Moller, 1978a). Low salinity may affect parasites directly, or

indirectly through its effects on less-tolerant host species essential to the
completion of parasite life cycles. Thulin (1984) recorded a decrease in the
parasite fauna of cod from 18 helminth and five copepod species on the
northwest coast of Sweden, where the salinity is 34 ppt, to only two
helminth species on the Baltic Sea coastline north of Stockholm, where
the salinity is 2 ppt. Engelbrecht (1958) showed that the ectoparasitic fauna
of freshwater fish is much reduced when the fish move into low salinity
coastal areas in the Baltic Sea. Osmanov and Yusopov (1985) found that
increased salinity in the Aral Sea, caused by human activity, resulted in a
reduction in both the numbers of species of fish parasites (from 215 in the
early 1970s to 37 in 1977), and the intensities of infection of the remaining
species. Moller (1987b) commented that similar effects of artificially raised
salinity on fish parasite faunas should become evident in Central European
sections of the Rivers Rhine and Weber that receive waste water from
salines. Moller (1981, 1984) found higher levels of disease in flounders
Pkutichthysflesus from the central estuaries of the River Weser and Elbe,
which experience wide variations in salinity due to tidal effects, than in
flounders from regions of more stable salinity upstream and downstream.
Thjs suggests that the unstable environment has an adverse effect on
these fish, rendering them more susceptible to disease and parasitic
infection.
Low oxygen concentrations can result from the introduction of fertilizers
from agriculture, sewage and other organically enriched pollutants. Para-
site eggs which sink to the anoxic bottom layers of a water body may be
particularly vulnerable; the eggs of some cestodes and leeches are known
to be very sensitive to oxygen depletion (Schleip et ul., 1937; Bauer, 1962).
Dorjes (1986) reported changes since the 1920s and 1930s in benthic
communities, which include the intermediate hosts of helminth parasites,
in the Helgoland region of the North Sea. Several species have dis-
appeared, others have decreased markedly, and some have undergone

large fluctuations in abundance.
The biotic factors that affect diseased conditions in fish (see Moller,
1985, 1987a) may also affect parasite numbers and distribution. Among
these factors are: competition or negative interaction between parasites; the
nutritional state of the host; spawning, which reduces resistance to para-
sites; and high host population densities, which favour parasite transmis-
sion. A knowledge of host migration patterns is important when
parasitological data are being assessed for the influence of pollutants.
Heavily parasitized hosts may behave differently from uninfected or
lightly infected individuals. The occurrence of some parasites is dependent
on host age, so it may be important to compare age-matched samples.
4. HELMINTH LIFE CYCLES (TABLES 6-13), TRANSMISSION

PROCESSES AND WATER QUALITY
About 200 fish helminth life cycles have been elucidated, and we there-
fore have a good knowledge of the basic transmission patterns for each
of the main taxonomic groups, namely Monogenea, Gyrocotylidea,
Amphilinidea, Caryophyllidea, Cestoda, Aspidogastrea, Digenea,
Nematoda and Acanthocephala. A transmission route is the pathway
through which a parasite progresses from one developmental stage to
the next and from one host to another. Most helminths have a range of
possible host species at certain life cycle stages, resulting in a number
of possible transmission routes. Associated with each transmission route
is a transmission window - the period during which transmission of the
parasite from one host to another can take place. The length of the
window depends on the concurrence of infective stages of the parasite
with hosts that are suceptible to infection. In some transmission routes
these windows are open virtually throughout the entire life span of the
host so that infection can take place at almost any time. In others the
windows are very narrow and transmission is limited to brief seasonal
periods. These narrow transmission windows represent weak links in
parasite life cycles at which routes through some host species may be
easily disrupted by changes in environmental conditions.
The Monogenea and some Gyrocotylidea have delicate free-living
ciliated larval stages in their direct form of life cycle, which involves
one host only (Tables 6 and 7). The larvae are also known to be short-
lived (up to 30 h) and therefore transmission from one host to another must
take place during a very brief period in the life cycle. Some nematodes (e.g.
some cucullanids) also have a life cycle involving only one host, but the
surface of sole,
and then Fish host - the common sole,
Solca solca
Egg hatches to release

oncomiracidium
Figure 1 The life cycle of the monogenean Entobdella soleae.
survival time of the second-stage larva in sea water can be up to 75 days

(Table 12).
The life cycles of cestodes and acanthocephalans maturing in fish are
more complicated than those of the groups referred to in the previous
lamellae of fish I
Fish host - the whiting,

Mcrlangius mcrlangus
1
attach; to host
Egg hatches to release

oncomiracidium.
Figure 2 The life cycle of the monogenean Diclidophora merlangi.
paragraph, in that the development time for the egg may be longer, they
may or may not include free-living larval stages, and intermediate hosts
(invertebrates and fish) are often implicated (Tables 8, 9 and 13). The
number of possible transmission routes is therefore greater. The free-living
- Marine rays eg.

ta, Raja naevus
oncomiracidium
(no host chemical
stimulus required for
Figure 3 The life cycle of the monogenean Dictyocotyle coeliaca.
stages (coracidia) of trypanorhynch tapeworms survive in sea water for

some days according to temperature: the survival time of Grillotia
erinaceus coracidia is at least 14 days at 10-16°C and 9-12 days at
16-20°C (unpublished observations) and coracidia of Lacistorhynchus
Table 6 Life-cycles in the Monogenea. Most vulnerable stages: eggs (oncomir-

acidium within is very sensitive to chemicals in the environment); free-living,
hatched oncomiracidium.
Species Key observations References
Diplectanum These two species are ovoviviparous Oliver (1976, 1978,
aequans 1982)
D. sciaenae
Entobdella soleae Eggs attach to sand particles and Keam (1986);
(Figure 1) remain on the bottom; hatch in 27 Keam and Macdonald
days at 13-17°C; host-hatching (1976)
factor in fish mucus stimulates
oncomiracidia to emerge
Acanthocotyle Eggs on the bottom where rays spend Keam (1967);
lobianchi much of their time resting Keam and Macdonald
(1976);
Macdonald (1974)
Diclidophora spp. Eggs are laid in bunches; 13 marine Frankland (1955);
(Figure 2) species known to require temperature Llewellyn (1957);
over 8°C during incubation; hatching Macdonald (1974,
related to host behaviour; life span of 1975)
oncomiracidia short, 10-13 h
Squulonchocotyle Hatching stimulated by host skin Euzet and Raibaut
torpedinus, mucus. Eggs fall to the bottom; ( 1960);
Microcotyle hatched oncomiracidia short-lived at Ktari (1969);
salpae, 18°C Llewellyn (1962)
Gastrocotyle
trachuri
Microcotyle Infection prevalence and intensity Noisy and Maillard
chrysophrii details given (1980);
Oliver (1984)
Atrispinum Infection of bass Dicentrarchus labrax Winch (1983)
labracis occurs in the fourth year
Dictyocotyle Found in coelom of rays, e.g. Raja Williams ( 1965);
coeliaca batis, R. naevus, R. radiata, less Keam (1970, 1975)
(Figure 3) frequently in R. fullonica. At 10°C
eggs hatch after 4-5 months; at 12°C
in lighvdark periods of 12 h each,
hatching begins after 96 days with
no daily hatching rhythm; host skin
and mucus do not stimulate hatching
Calicotyle kroyeri Found in the rectal gland and cloaca Williams (1965);
of rays, e.g. Raja radiata, R . batis, Keam (1987)
R . fullonica
Table 7 Life cycles in the Gyrocotylidea. Parasites of the spiral valve of

chimaeroid fish; life cycle may be direct and a tissue-dwelling post-larval stage
may be normal part of the life cycle (Colin et al., 1986; Williams et al., 1987).
Most vulnerable stage: free-living lycophore (decacanth) larva.
~~
Gyrocotyle rugosa Eggs hatch within a few seconds of Manter (195 1)

being laid
Gyrocotyle Eggs need three weeks at about 12°C Simmons (1974)
fimbriata for hatching
Gyrocotyle urna At 6°C eggs hatch after 57 .days; Xylander (1986, 1989)
average survival time of laiva
24.5 h; maximum survival time,
72 h; freshly hatched larvae swim
>20 X faster than larvae 24 h old
Table 8 Life cycles in the Spathebothriidea. No free-swimming stage in the life

cycle. Eggs operculate and with filaments for attaching to seaweed before ingestion
by gammarids. Adults occur in salmonids, gadoids, sturgeons and flatfish. Adult
worms may be the most vulnerable.
Bothrimonus sp. Larval stages (plerocercoids) in Stark (1965);
gammarids; these develop precociously Sandeman and Burt
from December to April, coinciding (1972)
with migration of salmon Salmo salar
to the sea in March
tenuis (= dollfusi) survive for 35 days at 11”C, 58 days at 15°C and 19 days
-at 19°C (Sakanari and Moser, 1985a). This longevity opens the opportunity
for several transmission routes to be utilized.
The life cycles of digeneans are even more complicated, involving from
one to four developmental stages in different host species plus a number of
alternative host species for some of these stages, giving rise to a large
number of possible transmission routes (Table 11). A digenean is usually
highly specific to its primary host (usually a mollusc), infecting only a
single species, or a group of closely related species, but may have a wide
range of alternative intermediate or definitive host species. Alternative host
species in helminth life cycles may be arranged along a continuum from
those to which the parasite is best adapted and which provide it with the
highest probability of successful transmission, to “dead-ends’’ or “eco-
logical &ks”, through which no further development is possible (Holmes,
1979).
An example of a cestode species with alternative hosts giving widely
plerocercoid
ingestcd with
second
intermediate
. host by elasmobranchs of
elasmobranch gnvid pmglottid
'
definitive host.
the genus R a j a (body acgment
and develops to containing eggs)
adult detaches and is
I
shed with host
filer ocer coi d 7

faecii
pmglottid breaks
up in water 10
~rocercoid innested
L i b copcpodiy
fish second pcrculurn (lid) -
intermediate host
'on time = 3-6
Coracidium - f m - \1 to rele.sc
t-'
coracidium
+;EJK 3
coracidium swimming luvd stage.
survival lime = m u . 9-11
drys n 16-ZO°C (McCuCay
Procercoid unpubl). d u least 14 days
in copepod first procercoid 10-16°C. (Bates 1987)
intermediate
host. eg.
Acarrfa.
PscudocaIanus.
Poracalanw ,
Tcmora.
Figure 4 The life cycle of the strobilate tapeworm, Grillotia erinaceus.
different probabilities of successful transmission is the trypanorhynch

Grillotia smuris-goru. This cestode infects a number of teleost fish inter-
mediate hosts, including the pelagic species mackerel Scomber scombrus
and scad Truchurus truchurus. It is highly unlikely, however, that pre-adult
Adult cestode
plero
inges
sccond
intermediate Definitive host -
host by the spur dog,
elasmobrancb gravid proglottid
definitive host. Squalus acanthias (body segment
and develops to containing eggs)
adult &tubes and 1s
sbcd w ~ t bhost
proglottid brcaks
up in water LO
proccrcoid ingested
with copcpod by
fish second
intermediate host
Egg ingested by benthic (bottorn-

living) copcpods. No free-swimming
coracidiurn stage, (according to the
view expressed by MacKenzie 1975)
Figure 5 The life cycle of the strobilate tapeworm, Gilquinia squali.
cestodes in these hosts will ever complete their life cycles because the
definitive host, the angel-shark Squatina squatina, is a bottom-living
ambush predator which is unlikely ever to prey on pelagic fish (MacKenzie
et af., 1984). Transmission routes through demersal fish hosts such as the
sea-breams (Sparidae) are likely to be more successful.
Table 9 Life cycles in the Eucestoda (strobilate tapeworms). Trypanorhyncha: in
some species eggs hatch to release ciliated coracidia. Most vulnerable stage: free-
swimming coracidium.
Grillotia erinaceus Coracidium infected copepods and Ruszkowski (1934)
(Figure 4) developed to procercoid in experiments
Lacistorhynchus Hatching and survival of coracidium dependent Sakanari and Moser
tenuis (= Lacisto- on temperature and salinity (eggs hatched after (1985a);
rhynchus dolljhJa 11 days in 50% sea water at 15OC). Mudry and Dailey
Coracidium infected copepods and developed (1971);
to a procercoid in experiments; the Moser (1981);
plerocercoid stage is found in > 60 teleost Young (1954)
species. In lab, plerocercoids from the
shinerperch Cymatogaster aggregata infected
leopard shark Triakis semifasciata. The
copepod Tigriopus californicus, mosquito fish
Gambusia aflnis and T. semifasciata were
suitable first and second intermediate and
definitive hosts
Parachristianella No free-swimming coracidium. Develops in Mudry and Dailey
monomegacantha copepod to form an immature plerocercus (1971)
probably directly infective to elasmobranchs
Gilquinia squali No free-swimming coracidium. Probably MacKenzie (1975)
(Figure 5 ) develops in benthic copepods. Plerocercoids
in humours of eye of whiting Merlangius
merlangus. Adult worms in spurdog Squalus
acanthias
Pseudophyllidea Life cycles of fresh water species best known
Eubothrium Marine form's preferred host is Salmo solar. Kennedy (1978a, b);
crassum Eggs are sensitive to salinity changes: Kuperman (1978)
need 5-20 ppt
Bothrioce halus Adults in turbot Psetta maxima; plerocercoids Davey and Peachey
gregariusf in the teleost Gobius minutus (= second (1968);
intermediate host); first intermediate host may Robert et al. (1990)
be a crustacean. Plerocercoids in first
intermediate host appear to be infective to
young turbot
Diphyllidea Adults parasitize elasmobranchs; intermediate hosts include molluscs,
gammarids and crabs. Adult worms may be the most vulnerable
Echinobothrium Metacestodes in Carcinus maenas. Adults in Dollfus (1964)
afine Raja clavata
Lecanicephalidea Very little useful information available on life cycles, but marine
and Tetraphyllidea species are very abundant
a Sakanari and Moser (1985a, b) called their material L. tenuis, but collected from
the Pacific coast of the USA, which means that it is really L. dollfusi, according to
Beveridge and Sakanari (1987).
Bothriocephalus scorpii was recognized as a species complex: Renaud et al.
(1983) and Renaud and Gabrion (1984) distinguished four species: B. scorpii, B.
gregarius, B. barbatus and B . funiculus.
I
pre-adult
ingesrcd by the
Fish host -
the thornback Lays eggs
ray with the ray, Raja clavata

crustacean host,
and develops
into adult
I Egg - operculate
I L J
Symonda (1972) comments on
the low prcvilcnce of this
sugc in N. wrvegfcvr from
Preadult larva chc Nonh Sea compared to
occurs encysted that recorded by Cunningbun
in Nephrops (1887) from the aunc lowtion transmission route to
nearly 100 y c u s earlier. crustacean host unknown
norvegicus
Egg is ingested dirtctly by the

crustacean ffcphrops norvegicus
Figure 6 The life cycle of the aspidogastrean Stichocotyle nephropis.
Some marine digenetic trematodes with fish intermediate hosts provide

good examples of narrow transmission windows. All infection of plaice
Pleuronectes platessa with Stephanostomum baccatum and Rhipidocotyle
sp. occurs in the first year of life, with S. baccatum only during June and
July and with Rhipidocotyle sp. only from about the middle of September
to the end of October (MacKenzie and Gibson, 1970). Transmission of
each of these parasites to plaice is therefore limited to a period of 6-8
fish, Lophius piscatorius
penetrates fish second
I intermediate host and

develops to metacercaria
I
penetrates mollusc
Af 4
Cercaria - free-
swimming, fork-
tailed larval stage via exhalent siphon of
mollusc. Several
hundred emitted per
day
Figure I The life cycle of the digenean Bucephaloides gracilescens.
weeks in a maximum host life span of over 20 years. Cercariae of Cercaria

doricha and Cercaria pythionike are released from the mollusc host during
a period of about three months in spring and early summer (Wright, 1956).
Fish become infected through ingesting the cercariae which then penetrate
metacercaria
ingested with
second
intermediate
-
Main definitive host - five
bearded rockling, Ciliutu
-
I
inhabits body cavity
of rockling, and
stomach of eel and
conger eel
I
I
host copepod and n u s f c f u . (also reported from eel
develops to adult Anguilla angurlla and conger eel retained in
Conger conger) worm in body cavity of
rockling. Eggs are
released when rockling
dies or is eaten, and
\broken down. 1
Metacercaria - devel
in copepod second
intermediate host eg.
Tigrropsrs brevrcornis Miracidium -
possibly occurs, but
cercarial body penetrates to

haemocoel of copepod via a I
penetrates mollusc ?
fsporoeyst - germinal sac,

occurs in the prosobranch
mollusc Gibbula umbilicalis.
cercariae emerge Cercariae develoo inside
Figure 8 The life cycle of the digenean Lecithochirium furcolabiatum.
the gut wall and encyst in the visceral cavity. All infection of herring
Clupea harengus takes place in their first summer of life, when they feed
on planktonic organisms of approximately the same size as these cercariae.
The fastest growing juvenile herring apparently progress to feeding on
Table 10 Life cycles in the Aspidogastrea. A small group poorly adapted to

parasitism; most use molluscs as the only host. Most vulnerable stage: free-
swimming cotylocidium.
Stichocotyle nephropis he-adults encyst in crustaceans e.g. Odhner (1910);
(Figure 6 ) Nephrops (transport/accidental hosts); MacKenzie (1964);
adults in rays Symonds (1972)
Taeniocotyle elegans Free-swimming cotylocidium hatches "honey and
(Syn. Macraspsis from operculate eggs Burreson (1987)
elegans)
Rugogaster hydrolagi Aciliate cotylocidium hatches from Schell (1973)
operculate eggs
larger organisms before the end of the period of cercarial emergence

(MacKenzie, 1985). Transmission to herring is therefore limited to a
period of 1-3 months in a maximum host life span of over 20 years. In a
year when herring growth rate is high and cercarial emergence is delayed,
levels of infection might be greatly reduced.
A good example of how human interference can drastically affect the
transmission of a parasite is given by Hanzelova (1992). In the year
following drainage of a freshwater reservoir in Czechoslovakia, the species
composition of the copepod community and the seasonal dynamics of
copepod numbers changed markedly. The copepods serve as intermediate
hosts for the fish tapeworm Proteocephalus neglectus and the effect on
transmission of this parasite in the year following drainage was that infec-
tion of copepods decreased by 95% and that of the fish definitive host by
97.5%.
Transmission failure of marine parasites are also recorded in the litera-
ture. Transmission of the cestodes G. smuris-gora and Lacistorhynchus
sp. to mackerel and herring respectively in two European study areas
was suddenly greatly reduced in 1979 and has not recovered since
(MacKenzie, 1987, and unpublished results). Transmission of the parasitic
copepod Pennella sp. to saury Cololabis saira in the Northwest Pacific
failed in 1985 and no infected saury were found in this area in 1986
(Nagasawa et al., 1988). However, the reappearance of the parasite in
1990 was reported by Honma and Imai (1991). The reasons for these
marine transmission failures are unknown, but are probably related to
natural environmental changes. It is probably significant that the study
areas referred to are close to the limits of the parasites' geographical
distributions. Populations at the fringes of a species' range are likely to
be the most vulnerable to even small changes in environmental conditions.
Table 11 Life cycles in the Digenea. The life cycles involve active penetration
of the mollusc first intermediate host by free-living miracidia and penetration of the
second intermediate host by free-living cercariae. These free-living stages are the
most vulnerable. Free-swimming miracidia are present in rhe life cycles of digen-
eans in the following families: Allocreadiidae, Bucephalidae, Derogenidae, Didy-
mozoidae, Gorgoderidae, Lecithasteridae, Lepocreadiidae and Opecoelidae. Free-
swimming cercariae are present in the life cycles of digeneans in the families
Hemiuridae, Fellodistomidae (Angel, 1971; K0ie, 1979a, 1980), Derogenidae
( K ~ i e ,1979b), Acanthostomidae (Maillard, 1976), Azygiidae (Stunkard, 1956)
and Bivesiculidae (Coil et al., 1965; Pearson, 1968). In the last two families,
cercariae are eaten by fish.
Proctoeces maculata This species may be ideal in view of the Freeman and
(syn. subtenuis) range of its life cycle strategies, which Llewellyn (1958)
(Fellodistomidae) include three or more generations of
sporocysts which produce cercariae in Bray and Gibson
spring-summer when water temperature (1980);
is high and labrid fish present, but only Bray (1983)
sporocysts in autumn when temperatures
fall and fish move offshore. In estuaries Aitken-Ander
of southeast England, sexually mature and Levin
adults are found in the lamellibranch (1985);
Scrobicularia plana Turner (1986)
Opechona Free-swimming miracidia penetrate Kdie (1975);
bacillaris Nassarius pygmaeus (first intermediate Bray and Gibson
(Lepocreadiidae host) and rediae produce cercariae which ( 1990)
emerge and penetrate ctenophores,
chaetognaths and medusae, but do not
encyst. Adult worms are found in the
teleost fish Cyclopterus lumpus and
Scomber scombrus
Derogenes varicus First intermediate hosts are molluscs of K d e (1979b)
- (Derogenidae) the genus Natica
Stephanostomum First intermediate host is the gastropod Koie (1978)
caducum Natica alderi. Rediae produce cercariae
(Acanthocolpidae) which emerge to encyst in gobies
(Pomatoschistus pictus, P. microps and
Chaparrudo flavescens), usually just
under epithelium of mouth. The final host
is the cod Gadus morhua
Podocotyle reflexa Free-living miracidia penetrate the KBie (1981);
(Opecoelidae) prosobranch Buccinum undatum; sporo- Gibson and Bray
cysts produce cotylocercous xiphidio- (1982)
cercariae which penetrate amphipods,
decapods and mysids as second inter-
mediate hosts. Adult worms are found in
the teleosts Gadus morhuu, Pholis
gunnellus, Rhinonemus cimbrius,
Cyclopterus lumpus and Zoarces viviparus
Table 11 Continued
~~ ~ ~

Paratimnia gobii First intermediate host is Abra ovata. Maillard (1976)
(Monorchiidae) Sporocysts give rise to gymnocephalous
cercariae which encyst in inhalent siphon
of lamellibranchs (Abra ovata, Cardium
glaucum). Adults are found in the teleost
Pomatoschistus microps
Bucephaloides First intermediate host is the mollusc Matthews (1974)
gracilescens Abra alba; metacercariae in gadoid fish;
. (Bucephalidae) definitive host is the angler fish Lophius
(Figure 7) piscatorius
Lecithochirium First intermediate host is the mollusc Matthews (1980,
furcolabiatum Gibbula umbilicalis, metacercariae in 1981a, b, 1982)
(Hemipidae) copepods, e.g. Tigriopsis brevicornis, main
(Figure 8) definitive host is the five-bearded rockling
Ciliata mustela, but adult also found in
eels Anguilla anguilla and conger eel
Conger conger
Zoogonoides Sporocysts produce tailless K ~ i (1976,
e 1983)
viviparus xiphidiocercariae which creep on the
(Zoogonidae) substrate using suckers. These penetrate Bray and Gibson
and encyst to form metacercariae in a (1986)
wide variety of second intermediate hosts
including brittle stars, polychaetes,
lamellibranchs, prosobranchs; in Danish
waters the most important second
intermediate host is Ophiura albida (Klie,
1976). Adult digeneans are found in flatfish
(plaice, flounder, dab, long rough dab) and a
number of other teleost fish, particularly
Callionymus (dragonets), Zeus, Cottidae
(sculpins), Blennidae (blennies) and
Gobiidae (gobies)
Ptychogonimus Sporocysts containing cercariae float and Palombi (1942)
megastomus are eaten by crustacean intermediate hosts
Aporocotyle Rediae produce cercariae which penetrate Klie (1982)
simplex flatfish hosts and infect the vascular system
(Skguinicolidae)
Bum ( 1 980) noted that the two dominant trematode species in winter
flounder in Jamaica Bay, New York, fluctuated greatly over a relatively
short period (10 weeks) in the summer, and regarded this as an example of
temperature-dependent parasite seasonality. The trematode Lepocreadium
Table 12 Life cycles in the Nematoda. Most require an intermediate host, but this
is not obligatory for some, e.g. the cucullanids. Some cucullanid eggs hatch to
release second-stage larvae. Other cucullanid and anisakid eggs are released non-
embryonated. The rhabdochonids and cystidicolids release partly or fully embryo-
nated eggs. Camallanids and philometrids are viviparous, releasing first-stage
larvae which behave so as to attract intermediate hosts. Survival of eggs, first- and
second-stage larvae, and rate of egg development are temperature dependent.
Cucullanus truttae First moult occurs in egg; egg hatches in Moravec, (1976,
7-8 days at 22-24°C. In some 1979, 1980)
populations, lampreys Lampetra planeri
are main, obligatory intermediate hosts.
Salmonids are definitive hosts
C. minutus First moult may occur in egg; egg hatches Gibson (1972)
in seven days at 19°C. Intermediate host
may be involved, but no firm evidence
yet; adults in flounder, Platichthys Jesus
C. heterochrous Egg hatches in seven days at 19°C; first Gibson (1972)
moult occurs in water; direct infection of
definitive host (flounder Platichthys Jesus)
by second-stage larvae, is likely
Pseudanisakis Adults found in elasmobranchs, e.g. Raja Uspenskaya
rotundata radiata; intermediate hosts are decapods (1960)
(Lithodes sp.); paratenic hosts are gadid
and pleuronectid fish
Ascarophis morhua Eggs are released fully embryonated, are Uspenskaya
ingested by crustaceans Carcinus maenas (1960);
and Pagurus sp. as intermediate hosts Poinar and
and hatch to moult twice. Final hosts are Thomas (1976)
cod Gadus morhua and haddock
Melanogrammus aeglejnus
Phiionem Gravid female expelled with roe at Uhazy (1977a, b)
oncorhynchi spawning, bursts in water to release first-
stage larvae which are ingested by cyclo- Platzer and
poid copepods (Cyclops bicuspidatus, Adams (1967)
C.b. thomasi). At 10°C L1 survive 25
days in sea water. Infective copepods are KO and Adams
eaten by fish definitive hosts ( 1969)
Oncorhynchus nerka and other salmonids.
Development times of larvae in definitive KO and Adams
and intermediate hosts given by authors (1969);
listed opposite. Maturation of host and Bashirullah
parasite synchronized so that both are ( 1973)
gravid at spawning grounds, perhaps host
hormones mediate parasite reproduction
Proleptus obtusus Crabs (Carcinus maenas, Eupagurus Lloyd (1928)
bernhardus) are intermediate hosts, and
elasmobranchs are final hosts
Table 12 Continued
Echinocephalus Adults parasitic in spiral valve of KO (1975);
SPP. elasmobranchs: E. sinensis in Aetabatus KO et al. (1975b);
jagellurn; E. uncinatus in Trygon, Millemann
Myliobatis, Balistes and other fish; E. (1963);
pseudouncinatus in Heterodontus Andrews et al.
francisa, Myliobatis californicus; E. (1988);
overstreeti in Heterodontus Anderson (1988);
portusjacksoni; larvae found in molluscan Pearse and Timm
intermediate hosts: bivalves (oysters and (1971)
abalone) and possibly sea urchins for E.
pseudouncinatus; bivalves (Crassostrea
gigas) for E. sinensis; Chlamys bifrons,
Pecten albus for E. overstreeti with oysters
as possible paratenic hosts; the gastropod
Hemrfuscus pugilinus for E. uncinatus;
Haliotis corrugata and possibly sea urchins
for E. pseudouncinatus
Table 13 Life cycles in the Acanthocephala. Life cycles involve amphipod,

ostracod or copepod intermediate hosts and fish definitive hosts. Some species
incorporate a transport (paratenic) host in the life cycle. Eggs are very resistant to
environmental conditions; hatch only after ingestion by intermediate host. Buron
and Golvan (1986) and Golvan and Buron (1988) listed the intermediate and
definitive fish hosts of acanthocephalans.
~~
Echinorhynchus Eggs hatch at 23°C; intermediate host: Olson and Pratt

lageniformis Corophium spinicorne, final host: (1971)
Platichthys stellatus
E. gadi Intermediate hosts are amphipods and Buron and Golvan
mysids; final host cod Gadus morhua ( 1986);
Golvan and Buron
(1988)
Paratenuisentis Hatches in 1-6 h at 22-25°C Samuel and
ambiguus Bullock (1981)
setiferoides fluctuated from 100%prevalence and mean intensity of 68.5

per fish to 14.3% prevalence and a mean intensity of 5.9 per fish. The
trematode Zoogonus lusius varied in prevalence and mean intensity from
57.7% and 1.4 per fish to 9.5% and 3.6 per fish. Bum (1980) remarked that
such variability overshadows all but the most extreme pollution effects. In
this example, looking at transmission processes to understand the causes of
the variability would be the first step towards understanding the significance
of such variation in applied studies.
Most of the examples given above involve host species which are
probably not essential for the survival of populations of these particular
parasites, but which may make some contribution to the parasite’s total
reproductive output. Plaice are considered less important than some other
species of flatfish in the life cycle of S. baccatum, herring less important
than sprats Sprattus sprattus in those of C. doricha and C . pythionike, and
mackerel less important than demersal fish in that of G. smuris-gora.
Because they tend to be particularly vulnerable to changes in environ-
mental conditions, narrow transmission windows associated with less
important hosts could provide a highly sensitive advance warning system
for aquatic pollution. A significant change in environmental conditions
may result in greatly reduced transmission or even in complete failure.
For instance, Siddall et al. (1993) found that miraoidia of digeneans using
Buccinum undutum as an intermediate host were susceptible to toxic effects
of trace metals, and they considered this to be the major factor in reducing
parasite prevalence along a pollution gradient. Conversely, the effects of
environmental stress on hosts might result in increased transmission
(Paperna, 1975; Skinner, 1982; Overstreet, 1988). A significant deviation
either way from a normal rate of transmission may be a warning of adverse
environmental conditions. Monitoring of parasite prevalence could be used
to assess long-term, chronic effects of pollution in coastal and estuarine
waters, to detect and characterize pollution incidents and to determine the
dispersal patterns of known contaminants, e.g. trace metals around dump
sites (Siddall et al., 1993). A parasite-based index of pollution may
ultimately be most valuable as part of a combined monitoring approach,
including the analysis of sediment contamination, infaunal community
composition and laboratory bioassays. An approach to the use of these
trailsmission processes in pollution monitoring is proposed below.
5. GUIDELINES AND PROCEDURES FOR SELECTING HOSTS AND

PARASITES
5.1. Guidelines
1. The marine biology and parasitology of the study area should have been
well researched over a period of at least 25 years.
2. Attention should be focused on host species known to be non-migratory
or to have small migratory movements and on juvenile fish for which
parts of the study area serve as nursery grounds. Some species of the
genus Raja are good examples in British waters (Steven, 1947). It is
advisable to avoid migratory fish or invertebrates which are known to
travel long distances unless the study area is known to cover their full
migratory range.
3. Parasites which have been well studied with regard to their ecology and
life cycles are preferable as sentinels. They should also be easily seen if
present, easily collected and identified and adult stages should be
capable of surviving in cooled filtered sea water until they have pro-
duced large numbers of eggs for storage and experimentation. Ideally an
, abundance of eggs should be available for storage in sea water at the
time a species is collected, as is the case with some monogeneans,

cestodarians and tapeworms.
4. In fish, ectoparasitic worms and adult worms of the gut lumen are
preferable as sentinels since they are likely to have greater contact
with the external environment throughout the life cycle.
5. Highly site-specific parasite species are most likely to be sensitive to
host tissue changes caused by pollutants.
6. A knowledge of the geographical distribution of the parasite species
under investigation is desirable since species at the limits of their
geographical range are most likely to be sensitive and vulnerable to
man-made environmental changes.
7. If a parasite species has a wide range of hosts it is advisable to identify
“dead end” hosts and narrow transmission windows in the life cycle.
8. Attention should be focused on hosts in which the particular parasite
species reaches high levels of infection.
9. It should be borne in mind that sibling parasite species which are
morphologically very similar may have different environmental
sensitivities.
5.2. Procedures
While applying these guidelines we adopted procedures for North Sea

survey work from 1990 to 1992 with a view to listing helminth species
with known life cycles and selecting from these a short-list of what we
considered to be the most potentially useful indicator species. The life
cycles of some of these species are illustrated in Figures 1-8. The
procedures adopted were as follows:
1. Compilation of a checklist of the parasitic worms of British marine fish.
2. Use of this checklist, a large number of publications and the National
Museum of Wales helminth collections dating from the 1950s to assess
which hosts and parasite species were likely to be abundant, easily

collected and easily identified.
3. Initially, focusing attention on about a dozen species.
4. Collecting at sampling sites where pollution was known to occur and,
for comparison, at other sites at increasing distance from these towards
less-polluted areas.
5. Investigating prevalence and mean intensity of selected species.
6. Noting an observed trend of parasitic worm species which were
previously abundant becoming very rare or absent in the North Sea.
-6. CONCLUSIONS
A review of the literature shows that changes in the levels of infection of

certain parasites of fish and aquatic invertebrates may give an early
indication of environmental changes that will eventually adversely affect
the majority of less sensitive organisms. These parasites can therefore be
used as early warning indicators of deteriorating water quality. Helminth
parasites are considered to be particularly useful in this respect because
many aquatic species have delicate free-living transmission stages that are
highly sensitive to environmental change, so they would be among the first
organisms to show decreasing populations in a polluted environment. On
the other hand, low levels of pollution may have a hormetic effect on some
fish ectoparasites, enhancing reproduction and causing a marked increase
in parasite populations. This sort of change could also be due to immuno-
suppression of a host which is more sensitive to pollution than the
parasite being monitored. Before any form of pollution can be shown
to be the cause of such changes, other biotic and abiotic factors (host
diet, temperature, salinity etc.) that may influence the host-parasite rela-
tionships must be considered and eliminated, and bioassays of host tissue
response to pollution should be carried out (e.g. metallothionein assay,
ethoxyresorufin-0-de-ethylase (EROD)activity) so that infection levels
may be assessed in relation to an unequivocal host response due to exposure
to pollution. Laboratory experiments should be undertaken to investigate
the response of parasite transmission stages to selected pollutants.
Guidelines are suggested for the selection of the most appropriate host-
parasite systems and the most vulnerable stages in parasite life cycles.
Promising sentinel species in the British marine parasite fauna are the
monogeneans Diclidophora merlangi, Dictyocotyle coeliaca and Entob-
della soleae, the strobilate tapeworms Gilquinia squali and Grillotia
erinaceus, the aspidogastrean Stichocotyle nephrops and the digeneans
Bucephaloides gracilescens and Lecithochirium furcolabiatum.
ACKNOWLEDGEMENTS
A generous grant from the Department of the Environment to the National

Museum of Wales supported this research. We are grateful to Dr Chris
Reid and Dr Mike Roberts at the Department of the Environment for their
help and advice during the project. We also appreciate the support of
Professor Peter Morgan, Keeper of Zoology, National Museum of Wales,
Professor M.F. Claridge, School of Pure and Applied Biology, University
of Wales College of Cardiff and Professor A.D. Hawkins, Director,
SOAFD Marine Laboratory, Aberdeen for their support in providing
excellent facilities for the work. Dr Andrew McCarthy assisted in the
preparation of Figures 1-8.
Aho, J.M., Gibbons, J.W. and Esch, G.W. (1976). Relationship between thermal
loading and parasitism in mosquitofish. In ‘!Thermal Ecology 11” (G.W. Esch
and R. W. McFarlane, eds), pp. 213-218. Technical Information Center, Energy
Research and Development Agency (CONF - 750425).
Aitken-Ander, P. and Levin, N.L. (1985). Occurrence of adult and developmental
stages of Proctoeces maculatus (Trematoda: Digenea) in the gastropod Crepidula
convexa. Transactions of the American Microscopical Society 104,250-260.
Anderson, J.W., Dixit, D.B., Ward, G.S. and Foster, R.S. (1977). Effects of
petroleum hydrocarbons on the rate of heart beat and hatching success of
estuarine fish embryos. In “Physiological Responses of Marine Biota to
Pollutants” (F.J. Vemberg, A. Calabrese, F.P. Thurberg and W.B. Vemberg,
eds), pp. 241-258. Academic Press, New York.
Anderson, J.W., Neff, J.M., Cox, B.A., Tatem, H.E. and Hightower, G.M. (1974a).
Characteristics of dispersions and water-soluble extracts of crude and refined oils
and their toxicity to estuarine crustaceans and fish. Marine Biology 27, 75-88.
Anderson, J.W., Neff, J.M., Cox, B.A., Tatem, H.E. and Hightower, G.M. (1974b).
The effects of oil on estuarine animals: toxicity, uptake and depuration, respira-
tion. In “Pollution of Marine Organisms” (F.J. Vemberg and W.B. Vemberg,
eds), pp. 288-310. Academic Press, New York.
Anderson, R.C. ( 1988). Nematode transmission patterns. Journal of Parasitology
74, 30-45.
Andrews, A.K., van Valin, C.C. and Stebbings, B. (1966). Some effects of
heptachlor on bluegills, Lepomis macrochirus. Transactions of the American
Fisheries Society 95, 297-303.
Andrews, R.H., Beveridge, I., Adams, M. and Baverstock, P.R. (1988). Identifica-
tion of life cycle stages of the nematode Echinocephalus overstreeti by allozyme
electrophoresis. Journal of Helminthology 62, 153-157.
Angel, L.M. (1971). Burnellus gen. nov. (Digenea: Fellodistomidae), the life
history of the type species, B. trichofurcatus (Johnston and Angel, 1940), and
a note on a related species, Tandanicola bancrofti Johnston, 1927, both from

Australian freshwater catfish, Tandanus tandanus. Parasitology 62, 375-384.
Anikieva, L.V. (1982). The influence of waste waters of Segezhsk paper-and-pulp
plant on the helminth fauna of fish in Lake Vygozero. In “Ekologiya paraziti-
cheskikh organizmov v biogeotsenozahk severa”, pp. 72-83. Karel’skii Filial
Akademii Nauk SSSR, Institut Biologii, Petrozavodsk, USSR (In Russian).
Arme, C. and Halton, D.W. (1972). Observations on the occurrence of Diclido-
phora merlangi (Trematoda: Monogenea) on the gills of whiting, Gadus
merlangus. Journal of Fish Biology 4, 27-32.
Aston, S.R., Thornton, I., Webb, J.S., Purves, J.V. and Milford, B.L. (1974).
Stream sediment composition, an aid to water quality assessment. Water, Air
and Soil Pollution 3, 321-325.
Auffret, M. (1988). Histopathological changes .related to chemical contamination
in Mytilus edulis from field and experimental conditions. Marine Ecology -
‘ Progress Series 46, 101-107.
Barszcz, C.A., Yevich, P.P., Brown, L.R., Yarbrough, J.D. and Minchen, C.D.
(1978). Chronic effects of three crude oils on oysters suspended in estuarine
ponds. Journal of Environmental Pathology and Toxicology 1, 879-895.
Bashirullah, A.K.M. (1973). Infeccion causada por Philonema sp. entre trucha y
salmon (Abstract). Acta Cientifica Venezolana 24, 57-58.
Bates, R.M. (1987). Aspects of the biology of trypanorhynch tapeworms and
investigations on their use as biological tags. Ph.D. thesis, The Open
University, UK, 371 pp.
Bauer, O.N. (1962). “The Ecology of Parasites of Freshwater Fish.” Israel
Program for Scientific Translations, Jerusalem, 2 15 pp.
Bauer, O.N. (1968). The influence of environmental factors on reproduction of
fish parasites. Fisheries Research Board of Canada Translation Series No 1099,
12 PP.
Beveridge, I. and Sakanari, J.A. (1987). Lacistorhynchus dollfusi sp. nov. (Cestoda:
Trypanorhyncha) in elasmobranch fishes from Australian and North American
coastal waters. Transactionsof the Royal Society of South Australia 3, 147-154.
Blanton, W.G. and Robinson, M.C. (1973). Some acute effects of low-boiling
petroleum fractions on the cellular structures of fish gills under field condi-
tions. In “The Microbial Degradation of Oil Pollution” (D.G. Ahearn and S.P.
‘Meyers, eds), pp. 265-273. Center for Wetland Resources, Louisiana State
University, Baton Rouge, Louisiana.
Boyce, N.P. and Behrens Yamada, S. (1977). Effects of a parasite, Eubothrium
salvelini (Cestoda: Pseudophyllidea), on the resistance of juvenile sockeye
Salmon, Oncorhynchus nerka, to zinc. Journal of the Fisheries Research Board
of Canada 34,706-709.
Bray, R.A. (1983). On the fellodistomid genus Proctoeces Odhner, 1911 (Digenea)
with brief comments on two other fellodistomid genera. Journal of Natural
History 17, 321-339.
Bray, R.A. and Gibson, D.I. (1980). The Fellodistomidae (Digenea) of fishes from
the northeast Atlantic. Bulletin of the British Museum (Natural History), Zoology
series 37, 199-293.
Bray, R.A. and Gibson, D.I. (1986). The Zoogonidae (Digenea) of fishes from the
northeast Atlantic. Bulletin of the British Museum (Natural History), Zoology
series 51, 127-206.
Bray, R.A. and Gibson, D.I. (1990). The Lepocreadiidae (Digenea) of fishes of the
northeast Atlantic: review of the genera Opechona Looss, 1907 and Prodisto-
mum Linton, 1910. Systematic Parasitology 15, 159-202.
Brown, A.F. and Pascoe, D. (1989). Parasitism and host sensitivity to cadmium: an
acanthocephalan infection of the freshwater amphipod Gammaruspulex. Journal
of Applied Ecology 26, 473487.
Brown, B.E. (1977). Effects of mine drainage on the River Hayle, Comwall. A.
Factors affecting concentrations of copper, zinc and iron in water, sediments and
dominant invertebrate fauna. Hydrobiologia 52, 22 1-223.
Bryan, G.W. and Hummerstone, L.G. (1973). Brown seaweed as an indicator of
heavy metals in estuaries in south-west England. Journal of the Marine
Biological Association of the United Kingdom 53, 705-720.
Bum, P.R. (1980). Pollution effects on fish parasites. Coastal Ocean Pollution
Assessment News 1, 3 4 .
Buron, I. and Golvan, Y.J. (1986). Les HBtes des Acanthoctphales I. Les H6tes
intermediares. Annales de Parasitologie Humaine et Comparke 61, 58 1-592.
Bychowsky, B.E. (1961). “Monogenetic Trematodes. Their Systematics and
Phylogepy.” English Translation by P.C. Oustinoff (W.J. Hargis, Jr, ed.).
American Institute of Biological Sciences, Washington, DC,627 pp.
Chubb, J.C. (1977). Seasonal occurrence of helminths in freshwater fishes. Part I.
Monogenea. Advances in Parasitology 15, 133-199.
Chubb, J.C. (1979). Seasonal occurrence of helminths in freshwater fishes. Part II.
Trematoda. Advances in Parasitology 17, 141-3 13.
Chubb, J.C. (1980). Seasonal occurrence of helminths in freshwater fishes. Part 111.
Larval Cestoda and Nematoda. Advances in Parasitology 18, 1-120.
Chubb, J.C. (1982). Seasonal occurrence of helminths in freshwater fishes. Part IV.
Cestoda, Nematoda, Acanthocephala. Advances in Parasitology 20, 1-292.
Coil, W.H., Reid, W.A. and Kuntz, R.E. (1965). Paucivitellosusfragilis gen. et sp.
nov. (Bivesiculidae: Digenea), a parasite of Chelon troscheli from Formosa.
Transactions of the American Microscopical Society 84, 365-368.
Cole, H.A. (1979). The assessment of sub-lethal effects of pollutants in the sea.
Philosophical Transactions of the Royal Society of London ( B ) 286, 399424.
Colin, J.A., Williams, H.H. and Halvorsen, 0. (1986). One or three gyrocotylideans
(Platyhelminthes) in Chimaera monstrosa (Holocephali)? Journal of Parasitology
72, 10-21.
Cone, D.K., Marcogliese, D.J. and Watt, W.D. (1993). Metazoan parasite commu-
nities of yellow eels (Anguilla rostrata) in acidic and limed rivers of Nova
Scotia. Canadian Journal of Zoology 71, 177-184.
Dabrowska, H. (1974). An attempt to evaluate the state of health of fish from the
Lyna and Walsza Rivers in connection to their pollution. Przeglad Zoologicny
18, 390-395.
Das, M.C. and Shrivastava, A.K. (1984). Fish mortality in Naini Tal Lake (India)
due to pollution and parasitism. Hydrobiological Journal 20, 60-64.
Davey, J.T. and Peachey, J.E, (1968). Bothriocephalus scorpii (Cestoda: Pseudo-
phyllidea) in turbot and brill from British coastal waters. Journal of the Marine
Biological Association of the United Kingdom 48, 335-340.
Dey, A.C., Kiceniuk, J.W., Williams, U.P., Khan, R.A. and Payne, J.F. (1983).
Long term exposure of marine fish to crude petroleum. 1. Studies on liver lipids
and fatty acids in cod (Gadus morhua) and winter flounder (Pseudopleuronectes
americanus). Comparative Biochemistry and Physiology 75, 93-101.
DiMichele, L. and Taylor, M.H. (1978). Histopathological and physiological
responses of Fundulus heteroclitus to naphthalene exposure. Journal of the

Fisheries Research Board of Canada 35, 1060-1066.
Dollfus, R.P. (1964). Sur le cycle tvolutif d’un cestode Diphyllide, Identification
de la larve chez Carcinus maenas (L. 1758), h6te intermediaire. Annales de
Parasitologie Humuine et Comparke 39, 235-24 1.
Dorjes, J. (1986). Langfristige Entwicklungstendenzen des Makrobenthos der
Deutschen Bucht. Data submitted by the Federal Republic of Germany to the
Scientific and Technical Working Group, International North Sea Conference,
1987.
Dzika, E. (1987). Annual occurrence dynamics of common monogeneans on the
gills of bream from the Lake Goslawskie (Poland). Acta Parasitologica Polonica
32, 121-137.
Eiras, J.C. (1987). Occurrence of Haemogregarina bigemina (Protozoa: Apicom-
plexa) in Blennius pholis (Pisces: Blenniidae) along the Portuguese west coast.
‘
Journal of Fish Biology 30, 597-603.
Engelbrecht, H. (1958). Untersuchungen iiber den Parasitenbefall der Nutzfische
im Greifswalder Bodden und Kleinen Haff. Zeitschrift f i r Fischerei 7,481-51 1.
Esch, G.W. (1982). Abiotic factors: an overview. In “Parasites -Their World and
Ours” (D.F. Mettrick and S.S. Desser, eds), pp. 279-288. Elsevier Biomedical
Press, Amsterdam.
Esch, G.W., Hazen, T.C., Dimock, R.V. and Gibbons, J.W. (1976). Thermal
effluent and the epizootiology of the ciliate Epistylis and the bacterium
Aeromonas in association with centrarchid fish. Transactions of the American
Microscopical Society 95, 687-693.
Ewe, H.E. and Esch, G.W. (1974). The effects of thermal effluents on the popula-
tion of helminth parasites in the largemouth bass, Micropterus salmoides. In
“Thermal Ecology” (J.W. Gibbons and R.R. Shority, eds), pp. 207-215. AEC
Symposium Series (CONF - 730505).
Euzet, L. and Raibaut, A. (1960). Le dtvelopment postlarvaire de Squulon-
chocotyle torpedinus (Price, 1942) (Monogenea, Hexbothriidae). Bulletin de la
SociCt6 Neuchdteloise des Sciences Naturelles 83, 101-108.
Evans, E.E., Kent, S.P., Attleberger, M.H., Seibert, C., Bryant, R.E. and Booth, B.
(I 965). Antibody synthesis in poikilothermic vertebrates. Annals of the New
York Academy of Sciences 126, 629-646.
Evans, N.A. (1982a). Effects of copper and zinc on the life cycle of Notocotylus
attenuatus (Digenea: Notocotylidae). International Journal for Parasitology 12,
363-369.
Evans, N.A. (1982b). Effect of copper and zinc upon the survival and infectivity of
Echinoparyphium recurvaturn cercariae. Parasitology 85, 295-303.
Evans, N.A. (1985). The influence of environmental temperature upon transmission
of the cercariae of Echinostoma liei (Digenea: Echinostomatidae). Parasitology
90,269-275.
Ewing, M.S.,Ewing, S.A. and Zimmer, M.A. (1982). Sublethal copper stress and
susceptibility of channel catfish to experimental infections with Ichthyophthirius
multifliis. Bulletin of Environmental Contamination and Toxicology 28,674-68 1.
Forstner, U. and Whittman, G.T.W. (1979). “Metal Pollution in the Aquatic
Environment. ” Springer-Verlag, Berlin, Heidelberg, 486 pp.
Frankland, H.M.T. (1955). The life history and bionomics of Diclidophoru
denticulata (Trematoda: Monogenea). Parasitology 45, 3 13-35 1.
Freeman, R.F.H. and Llewellyn, J. (1958). An adult digenetic trematode from an
invertebrate host: Proctoeces subtenuis (Linton) from the lamellibranch
Scrobicularia plana (da Costa). Journal of the Marine Biological Association

of the United Kingdom 37, 435457.
Fries, C.R. (1986). Effects of environmental stressors and immunosuppressants on
immunity in Fundulus heteroclitus. American Zoologist 26, 27 1-282.
Gardner, G.R. (1975). Chemically induced lesions in estuarine or marine teleosts.
In “The Pathology of Fishes” (W.E. Ribelin and G. Migaki, eds), pp. 657-694.
The University of Wisconsin Press, Madison, Wisconsin.
Gibson, D.I. (1972). Contributions to the life-histories and development of
Cucullanus minutus Rudolphi, 1819 and C. heterochrous Rudolphi, 1802
(Nematoda: Ascaridida). Bulletin of the British Museum (Natural History),
Zoology series 22, 151-1 70.
Gibson, D.I. and Bray, R.A. (1982). A study and reorganization of Plagioporus
Stafford, 1904 (Digenea: Opecoelidae) and related genera, with special reference
to forms from European Atlantic waters. Journal of Natural History 16,
529-559.
Golvan, Y.J. and Buron, I. de (1988). Les HBtes des AcanthocCphales 11. Les
HBtes Definitifs. 1. Poissons. Annales de Parasitologie Humaine et Comparke
63, 349-375.
Granath, W.O. Jr and Esch, G.W. (1983a). Temperature and other factors that
regulate the composition and infrapopulation densities of Bothriocephalus
acheilognathi (Cestoda) in Gambusia afinis (Pisces). Journal of Parasitology
69, 1116-1 124.
Granath, W.O. Jr and Esch, G.W. (1983b). Seasonal dynamics of Bothriocephalus
acheilognathi in ambient and thermally altered areas of a North Carolina cooling
reservoir. Proceedings of the Helminthological Society of Washington 50,
205-2 18.
Grizzle, J.M. (1986). Lesions in fish captured near drilling platforms in the Gulf of
Mexico. Marine Environmental Research 18, 267-276.
Guth, D.J., Blankespoor, H.D. and Cairns, J. Jr (1977) Potentiation of zinc stress
caused by parasitic infection of snails. Hydrobiologia 55, 225-229.
Haensly, W.E., Neff, J.M., Sharp, J.R., Moms, A.C., Bedgood, M.F. and Boem,
P.D. (1982). Histopathology of Pleuronectes platessa L. from Aber Wrac’h and
Aber Benoit, Brittany, France: long-term effects of the Amoco Cadiz crude oil
spill. Journal of Fish Diseases 5 , 365-391.
Hansen, M.A., Fernandes, G. and Good, R.A. (1982). Nutrition and immunity.
Annual Review of Nutrition 2, 151-177.
Hanzelova, V. (1992). Proteocephalus neglectus as a possible indicator of changes
in the ecological balance of aquatic environments. Journal of Helminthology 66,
17-24.
Hawkes, J.W. (1977). The effects of petroleum hydrocarbon exposure on the
structure of fish tissues. In “Fate and Effects of Petroleum Hydrocarbons in
Marine Ecosystems and Organisms” (D.A. Wolfe, ed.), pp. 115-128. Pergamon
Press, New York.
Hirschfield, M.F., Morin, R.P. and Hepner, D.J. (1983). Increased prevalence of
larval Eustrongylides (Nematoda) in the mummichog, Fundulus heteroclitus L.,
from the discharge canal of a power plant in Chesapeake Bay. Journal of Fish
Biology 23, 135-142.
Hodgins, H.O., McCain, B.B. and Hawkes, J.W. (1977). Marine fish and inverte-
brate diseases, host disease resistance, and pathological effects of petroleum. In
“Effects of Petroleum on Arctic and Subarctic Marine Environments and
Organisms. Volume 2. Biological Effects” (D.C. Malins, ed.), pp. 95-173.

Academic Press, New York.
Hoffman, G.L. and Putz, R.E. (1964). Studies on Gyrodactylus macrochiri n. sp.
(Trematoda: Monogenea) from Lepomis macrochirus. Proceedings of the
Helminthological Society of Washington 31, 76-82.
Hoglund, J. and Thulin, J. (1989). Thermal effects on the seasonal dynamics of
Paradiplozoon homoion (Bychowsky & Nagibina, 1959) parasitizing roach,
Rutilus rutilus (L.). Journal of Helminthology 63, 93-101.
Holmes, J.C. (1979). Parasite populations and host community structure. In “Host-
Parasite Interfaces” (B.B. Nickol, ed.), pp. 27-46. Academic Press, London.
Honma, T. and Imai, Y. (1991). Reoccurrence of Pennella sp. (Copepoda) on the
saury, Cololaibis saira, in 1990. Scientifrc Reports of Hokkaido Fisheries
Experimental Station no. 37, 67-69.
’
Janiszewska, J. (1939). Studien uber die Entwicklung und die Lebensweise der
parasitischen Wiirmer in der Flunder (Pleuronectes Jesus L.). Mtmoires de
I’Acadkmie Polonaise des Sciences et des Lettres, SBrie B, 14, 1-68.
Joo, C.J.(1988). Changing patterns of infection with digenetic larval trematodes
from fresh-water fish in River Taewha, Kyongnam Province. Korean Journal of
Kamiso, H.N. and Olson, R.E. (1986). Host-parasite relationships between Gyro-
dactylus stellatus (Monogenea: Gyrodactylidae) and Parophrys vetulus (Pleuro-
nectidae -English sole) from coastal waters of Oregon. Journal of Parasitology
72, 125-129.
Kearn, G.C. (1967). The life-cycles and larval development of some acantho-
cotylids (Monogenea) from Plymouth rays. Parasitology 57, 157-167.
Kearn, G.C. (1970). The oncomiracidia of the monocotylid monogeneans
Dictyocotyle coeliaca and Calicotyle kroyeri. Parasitology 61, 153-1 60.
Kearn, G.C. (1975). Hatching in the monogenean parasite Dictyocotyle coeliaca
from the body cavity of Raja naevus. Parasitology 70, 87-93.
Kearn, J.C. (1986). The eggs of monogeneans. Advances in Parasitology 25,
175-273.
Keam, G.C. (1987). The site of development of the monogenean Calicotyle
kroyeri, a parasite of rays. Journal of the Marine Biological Association of the
United Kingdom 67, 77-87.
Kearn, G.C. and Macdonald, S. (1976). The chemical nature of host hatching
factors in the monogenean skin parasites Entobdella soleae and Acanthocotyle
lobianchi. International Journal for Parasitology 6, 457466.
Kennedy, C.R. (1978a). Studies on the biology of Eubothrium salvelini and E.
crassum in resident and migratory Salvelinus alpinus and Salmo trutta and in S.
salar in North Norway and the islands of Spitsbergen and Jan Mayen. Journal of
Fish Biology 12, 147-162.
Kennedy, C.R. (1978b). The biology, specificity and habitat of the species of
Eubothrium (Cestoda: Pseudophyllidea), with reference to their use as biological
tags: a review. Journal of Fish Biology 12, 393-410.
Khan, R.A. (1987a). Effects of chronic exposure to petroleum hydrocarbons on
two species of marine fish infected with a hemoprotozoan Trypanosoma
murmanensis. Canadian Journal of Zoology 65, 2703-2709.
Khan, R.A. (1987b). Crude oil and parasites of fish. Parasitology Today 3.99-100.
Khan, R.A. (1990). Parasitism in marine fish after chronic exposure to petroleum
hydrocarbons in the laboratory and to the Exxon Valdez oil spill. Bulletin of
Environmental Contamination and Toxicology 44,759-763.
Khan, R.A. and Kiceniuk, J. (1983). Effects of crude oil on the gastrointestinal
parasites of two species of marine fish. Journal of Wildlife Diseases 19,
253-258.
Khan, R.A. and Kiceniuk, J. (1984). Histopathological effects of crude oil on
Atlantic cod following chronic exposure. Canadian Journal of Zoology 62,
2038-2043.
Khan, R.A. and Kiceniuk, J.W. (1988). Effect of petroleum aromatic hydro-
carbons on monogeneids parasitizing Atlantic cod, Gadus morhua L. Bulletin
of Environmental Contamination and Toxicology 41,94-1 00.
Khan, R.A. and Thulin, J. (1991). Influence of pollution on parasites of aquatic
animals. Advances in Parasitology 30, 20 1-238.
Kiceniuk, J. and Khan, R.A. (1983). Toxicology of chronic crude oil exposure:
sublethal effects on aquatic organisms. In “Aquatic Toxicology” (J.O. Nraigu,
ed.), pp. 425-536. John Wiley, New York.
Kiceniuk, J.W. and Khan, R.A. (1987). Effect of petroleum hydrocarbons on
Atlantic cod, Gadus morhua, following chronic exposure. Canadian Journal of
Zoology 65, 490-494.
Kiceniuk, J.W., Khan, R.A., Dawe, M. and Williams, U. (1982). Examination of
interaction of trypanosome infection and crude oil exposure on hematology of
the longhorn sculpin (Myoxocephalus octodecemspinosus). Bulletin of
Environmental Contamination and Toxicology 28, 435-438.
KO, R.C. (1975). Echinocephalus sinensis n. sp. (Nematoda: Gnathostomatidae)
from the ray (Aetobatus jlagellum) in Hong Kong, Southern China. Canadian
Journal of Zoology 53, 490-500.
KO, R.C. and Adams, J.R. (1969). The development of Philonema oncorhynchi
(Nematoda: Philometridae) in Cyclops bicuspidatus in relation to temperature.
Canadian Journal of Zoology 47, 307-3 12.
KO, R.C., Morton, B. and Wong, P.S. (1975a). Preliminary studies on Echino-
cephalus sinensis (Nematoda: Spinuoidea), an unusual parasite from the oyster
Crassostrea gigas Thunberg, 1793 (Mollusca: Bivalvia) in Hong Kong. In
“Symposium Papers of the Pacific Science Association Special Symposium on
Marine Sciences” (B. Morton, ed.), pp. 66-70. PSA. Hong Kong.
KO, R.C., Morton, B. and Wong, P.S. (1975b). Prevalence and histopathology
of Echinocephalus sinensis (Nematoda: Gnathostomatidae) in natural and
experimental hosts. Canadian Journal of Zoology 53,550-559.
K@ie, M. (1975). On the morphology and life history of Opechona bacillaris
(Molin, 1859) Looss, 1907 (Trematoda, Lepocreadiidae). Ophelia 13,63-86.
Kflie, M. (1976). On the morphology and life history of Zoogonoides viviparus
(Olsson, 1868) Odhner, 1902 (Trematoda, Zoogonidae). Ophelia 15, 1-14.
Klie, M. (1978). On the morphology and life history of Stephanostomum
caducum (Looss, 1901) Manter, 1934 (Trematoda, Acanthocolpidae). Ophelia
17, 121-133.
Kgiie, M. (1979a). On the morphology and life history of Monascus (= Haplocladus)
jliformis (Rudolphi, 1819) Looss, 1907 and Steringophorus firciger (Olsson,
1868) Odhner, 1905 (Trematoda, Fellodistomidae). Ophelia 18, 113-1 32.
Kflie, M. (1979b). On the morphology and life history of Derogenes varicus
(Muller, 1784) Looss, 1901 (Trematoda, Hemiuridae). Zeitschrifi fur Parasiten-
kunde 59,67-78.
Kgiie, M. (1980). On the morphology and life history of Steringotrema pagelli
(van Beneden, 1871) Odhner, 1911 and Fellodistomum fellis (Olsson, 1868)
Nicoll, 1909 [syn. S . ovacutum (Lebour, 1908) Yamaguti, 19531 (Trematoda,

Fellodistomidae). Ophelia 19, 215-236.
Kgie, M. (1981). On the morphology and life history of Podocotyle refrexa (Creplin,
1825) Odhner, 1905, and a comparison of its developmental stages with those of
P. atomon (Rudolphi, 1802) Odhner, 1905 (Trematoda, Opecoelidae). Ophelia
20, 1743.
Kgie, M. (1982). The redia, cercaria and early stages of Aporocotyle simplex
Odhner, 1900 (Sanguinicolidae) - a digenetic trematode which has a
polychaete annelid as the only intermediate host. Ophelia 21, 115-145.
Kgie, M. (1983). Digenetic trematodes from Limanda limanda (L.) (Osteichthyes,
Pleuronectidae) from Danish and adjacent waters, with special reference to their
life histories. Ophelia 22, 201-228.
Koskivaara, M., Valtonen, E.T. and Prost, M. (1991). Dactylogyrids on the gills
of roach in central Finland: features of infection and species composition.
Kostarev, G.F. (1980). The effects of pollution on the parasite fauna in the
Kama reservoirs. In “Biologicheskie resursy vodoemov zapadnogo Urala”, pp.
147-153. Mezhvuzovskii Sbornik Nauchnykh Trudov. Permskii Gosudarstvennyi
Universitet, USSR (In Russian).
Ktari, M.H. (1969). RBcherches sur l’anatomie et la biologie de Microcotyle salpae
Parona et Perugia, 1890 parasite de Box salpa L. (TCltostBen). Annales de
Parasitologie Humaine at ComparCe 441, 425-439.
Kuperman, B.I. (1978). Aspects of the life cycle and biology of cestodes from
Salmonidae in the Kamchatka. Biologiya Morya, Vladivostok 4, 53-60 (In
Russian).
Lehtinen, K.J., Notini, M. and Landner, L. (1984). Tissue damage and parasite
frequency in flounders, Platichthys fresus L. chronically exposed to bleached
kraft pulp mill effluents. Annales Zoologica Fennici 21, 23-28.
Lester, R.G.J. and Adams, J.R. (1974). Gyrodactylus alexanderi: reproduction,
mortality, and effect on its host Gasterosteus aculeatus. Canadian Journal of
Zoology 52, 827-833.
Liao, H. and Shih, L. (1956). Contribution to the biology and control of
Bothriocephalus gowkongensis Yeh, a tapeworm parasitic in young grass
carp (Ctenopharyngodon idellus). Acta Hydrobiologica Sinica 2, 129-1 85 (In
Chinese).
Llewellyn, J. (1957). The larvae of some monogenetic trematode parasites of
Plymouth fishes. Journal of the Marine Biological Association of the United
Kingdom 36, 243-259.
Llewellyn, J. (1962). The life histories and population dynamics of monogenean
gill parasites of Trachurus trachurus (L.). Journal of the Marine Biological
Association of the United Kingdom 42, 587-600.
Lloyd, J.H. (1928). On the life-history of the common nematode of the dogfish
(Scyllium canicula). Annals and Magazine of Natural History, Series 10, 1,
7 12-7 15.
Macdonald, S. (1974). Host skin mucus as a hatching stimulant in Acanthocotyle
lobianchi, a monogenean from the skin of Raja spp. Parasitology 68, 331-338.
Macdonald, S. (1975). Hatching rhythms in three species of Diclidophora
(Monogenea) with observations on host behaviour. Parasitology 71, 2 11-228.
MacKenzie, K. (1964). Stichocotyle nephropis Cunnungham, 1887 (Trematoda) in
Scottish waters. Annals and Magazine of Natural History, Series 13,6,505-506.
MacKenzie, K. (1975). Some aspects of the biology of the plerocercoid of
Gilquinia squali Fabricius 1794 (Cestoda: Trypanorhyncha). Journal of Fish

Biology 7,321-328.
MacKenzie, K. (1985).The use of parasites as biological tags in population studies
of herring (Clupea harengus L.) in the North Sea and to the north and west of
Scotland. Journal du Conseil. Conseil international pour l'exploration de la mer
42,3364.
MacKenzie, K. (1987).Long-term changes in the prevalence of two helminth
parasites (Cestoda: Trypanorhyncha) infecting marine fish. Journal of Fish
Biology 31,83-88.
MacKenzie, K. and Gibson, D.I. (1970).Ecological studies of some parasites of
plaice Pleuronectes platessa L.and flounder Platichthysflesus L. Symposia of the
British Society of Parasitology 8, 1-42.
MacKenzie, K., McVicar, A.H. and Waddell, I.F. (1976).Some parasites of plaice
Pleuronectesplntessa L. in three different farm environments. Scottish Fisheries
Research Report (4), 1-14.
MacKenzie, K., Smith, R.M. and Williams, H.H. (1984).Aspects of the biology
of Grillotia angeli Dollfus, 1969 (Cestoda: Trypanorhyncha) and its use as a
biological tag for mackerel and other fish. International Council for the
exploration of the Sea C.M.l984/H: 53, 7 pp. (mimeo).
Maillard, C. (1976). Distomatoses de poissons en milieu lagunaire. Thesis,
Universite des Sciences et Techniques du Languedoc, Montpellier, France,
383 pp.
Manter, H.W. (1951).Studies on Gyrocotyle rugosa Diesing, 1850,a cestodarian
parasite of the elephant fish, Callorhynchus milii. Zoology Publications from
Victoria University College, Wellington, New Zealand 17, 1 1 pp.
Margolis, L., Esch, G.W., Holmes, J.C., Kuris, A.M. and Schad, G.A. (1982).The
use of ecological terms in parasitology (report of an ad hoc committee of the
American Society of Parasitologists). Journal of Parasitology 68, 131-133.
Matthews, B.F. (1980). Cercaria vaullegeardi Pelseneer, 1906 (Digenea:
Hemiuridae); the daughter sporocyst and emergence of the cercaria. Parasitology
81,6169.
Matthews, B.F. (198la). Cercaria vaullegeardi Pelseneer, 1906 (Digenea:
Hemiuridae); development and ultrastructure. Parasitology 83,575-586.
Matthews, B.F. (198lb). Cercaria vaullegeardi Pelseneer, 1906 (Digenea:
Hemiuridae); the infection mechanism. Parasitology 83,587-593.
Matthews, B.F. (1982). The biology of the British Hemiuridae. PhD Thesis,
Plymouth Polytechnic, UK, 163 pp.
Matthews, R.A. (1974).The life-cycle of Bucephaloides gracilescens (Rudolphi,
1819)Hopkins, 1954 (Digenea: Gasterostomata). Parasitology 68,1-12.
May, R.M. (1988).How many species are there on earth? Science 241,1441-1449.
McCain, B.B., Hodgins, H.O., Gronland, W.D., Hawkes, J.W., Brown, D.W.,
Meyers, M.S. and Vandermeulen, J.H. (1978). Bioavailability of crude oil
from experimentally oiled sediments to English sole (Parophrys vetulus), and
pathological consequences. Journal of the Fisheries Research Board of Canada
35, 657-664.
McDonald, J.H. and Koehn, R.K. (1988).The mussels Mytilus galloprovincialis
and M. trossulus on the Pacific coast of North America. Marine Biology 99,
1 1 1-1 18.
McIntyre, A.D. and Pearce, J.B. (eds) (1980).Biological effects of marine pollu-
tion and the problems of monitoring. Rapports et proces-verbaux Rtunion.
Conseil permanent international pour l'exploration de la mer 179, 1-346.
McVicar, A.H. (1986). The use of fish pathology in programmes to monitor marine
contaminants. In “Report of the ICES Workshop in the Use of Pathology in
Studies of the Effects of Contaminants” (J. Thulin, ed.), pp. 58-64. International
Council for the Exploration of the Sea CM 1986/E: 40, Ref. F.
Millemann, R.E. (1963). Studies on the taxonomy and life history of echino-
cephalid worms (Nematoda: Spiruroidea) with a complete description of
Echinocephalus pseudouncinatus Millemann, 1951. Journal of Parasitology
49,754-764.
Mohan, C.V. and Sommerville, C. (1988). Effect of cadmium on susceptibility and
immune response of common carp to the protozoan Ichthyophthirius multijiliis
(Abstract). Programme and Abstracts of the Vrh European Multicolloquium of
Parasitology, 107.
Moller, H. (1976). Reduction of the intestinal fauna of marine fishes in captivity.
Journal of the Marine Biological Association of the United Kingdom 56,
781-785.
Moller, H. (1978a), The effects of salinity and temperature on the development and
survival of fish parasites. Journal of Fish Biology 12, 31 1-324.
Moller, H. (1978b). Ecological effects of cooling water of a power plant at
Kiel Fjord. Berichte der Deutschen Wissenschaflichen Kommission fur
Meeresforschung 26, 117-1 30.
Moller, H. (1981). Fish diseases in German and Danish coastal waters in
summer 1980. Berichte der Deutschen Wissenschaftlichen Kommission fur
Meeresforschung 29, 1-16.
Moller, H. (1984). Dynamics of fish diseases in the lower Elbe River. Helgolander
Meeresuntersuchungen 37, 3 8 9 4 13.
Moller, H. (1985). A critical review of the role of pollution as a cause of fish
diseases. In “Fish and Shellfish Pathology” (A.E. Ellis, ed.), pp. 169-182.
Academic Press, London.
Moller, H. (1987a). Pollution and parasitism in the aquatic environment. Inter-
national Journal for Parasitology 17, 353-362.
Moller, H. (1987b). The marine ecologist - scientist or advocate of nature?
Marine Pollution Bulletin 18,267-270.
Moravec, F. (1976). Occurrence of the encysted larvae of Cucullanus truttae
(Fabricius, 1794) in the brook lamprey, Lampetra planeri (Bl.). Scripta
Facultatis Scientiarum Naturalium Universitas Purkynianae Brunensis Biologia
I, 6, 17-20.
Moravec, F. (1979). Observations on the development of Cucullanus (Truttaedac-
nitis) truttae (Fabricius, 1794) (Nematoda: Cucullanidae). Folia Parasitologica
26, 295-307.
Moravec, F. (1980). Biology of Cucullanus truttae (Nematoda) in a trout stream.
Folia Parasitologica 27, 2 17-226.
Moser, M. (1981). Parasitological survey of San Francisco Bay - Delta area.
Striped bass and other select species. Report to California State Water Resources
Control Board, Standard Agreement 9-100-400-0, 114 pp.
Mudry, D.R. and Dailey, M.D. (1971). Postembryonic development of certain
tetraphyllidean and trypanorhynchan cestodes with a possible alternative
life cycle for the order Trypanorhyncha. Canadian Journal of Zoology 49,
1249-1 253.
Munkittrick, K.R. and Dixon, D.G. (1988). Growth, fecundity, and energy stores of
white sucker (Catostomus commersoni) from lakes containing elevated levels of
PARASITES AS INDICATORS OF WATER OUALITY 139
copper and zinc. Canadian Journal of Fisheries and Aquatic Sciences 45,
1355-1 365.
Nagasawa, K., Imai, Y. and Ishida, K. (1988). Long-term changes in the
population size and geographical distribution of Pennella sp. (Copepoda) on
the saury, Cololabis saira, in the western North Pacific Ocean and adjacent
seas. In “Biology of Copepods” (G.A. Boxshall and H.K. Schminke, eds), pp.
57 1-578. Kluwer Academic Publishers, Dordrecht, Belgium.
Noisy, D. and Maillard, C. (1980). Microhabitat branchial preferentiel de Micro-
cotyle chrysophrii van Beneden et Hesse, 1863 (Monogenea, Microcotylidae),
parasite de la Daurade (Sparus aurata L., 1758). Annales de Parasitologie
Humaine et Comparte 55, 3340.
Odhner, T. (1910). Stichocotyle nephropis J.T. Cunningham, ein aberranter
Trematode der Digenefamilie Aspidogastridae. Kungliga Svenska Vetenskapsa-
kademiens Handlingar 45, 1-16.
Oliver, G. (1976). New observations on the biology and ecology of some
Diplectanidae (Monogenea, Monopisthocotylea). Trudy Biologo-Pochvennogo
Instituta (Issledovaniya monogenticheskikh sosal’shchikov), Novaya Seriya 34,
104-109 (In Russian).
Oliver, G. (1978). Description de deux cas d’ovoviviparite chez les Diplectanidae
Bychowsky, 1957 (Monogenea, Monopisthocotylea). Zeitschrift fur Para-
sitenkunde 57, 247-250.
Oliver, G. (1982). Quelques aspects de la sp6cificitC parasitaire chez les
Diplectanidae Bychowsky, 1957 (Monogenea, Monopisthocotylea). Mtmoires
du Muskum nationale dhistoire naturelle, Serie A, Zoologie 123,295-301.
Oliver, G. (1984). Microcotyle chrysophrii van Beneden et Hesse, 1863 (Mono-
genea, Polyopisthocotylea, Microcotylidae) parasite de Sparus aurata Linnaeus,
1758 (Teleostei, Sparidae) dans les etangs littoraux du Languedoc-Rousillon
(France). Bulletin de la Socittt zoologique de France 109,113-1 18.
Olson, R.E. and Pratt, I. (1971). The life cycle and larval development of Echino-
rhynchus lageniformis Ekbaum, 1938 (Acanthocephala: Echinorhynchidae).
Journal of Parasitology 57, 143-149.
O’Neill, J.G. (1981). The humoral immune response of Salmo trutta L. and
Cyprinus carpio L. exposed to heavy metals: Journal of Fish Biology 19,
297-306.
Osmanov, S.O. and Yusopov, 0. (1985). Influence of the increasing salinity of the
Aral Sea on the fish parasite fauna. Parazitologicheskii Sbornik 33, 1 4 4 3 (In
Russian).
Overstreet, RM. (1988). Aquatic pollution problems; southeastern US coasts:
histopathological indicators. Aquatic Toxicology 11, 2 13-239.
Overstreet, R.M. (1993). Parasitic diseases of fishes and their relationship with
toxicants and other environmental factors. In “Pathobiology of Marine and
Estuarine Organisms” (J.A. Couch and J.W. Fournie, eds.), pp. 111-156. CRC
Press Inc., Boca Raton, Florida.
Overstreet, R.M. and Howse, H.D. (1977). Some parasites and diseases of estuarine
fishes in polluted habitats of Mississippi. Annals of the New York Academy of
Sciences 298,427462.
Palombi, A. (1942). I1 ciclo biologic0 di Ptychogonimus megastoma (Rud.),
Osservazioni sulla morfoligica e fisiologia delle forme larvali e considerazioni
filogenetiche. Rivista Parassitologia 6, 1 17-172.
Paperna, I. (1975). Parasites and diseases of the grey mullet (Mugilidae) with
special reference to the seas of the near East. Aquaculture 5, 65-80.
Paperna, I. and Overstreet, R.M. (1981). Parasites and diseases of mullets (Mugi-
lidae). In “Aquaculture of Grey Mullets” (O.H. Oren, ed.), pp. 41 1-493. Inter-
national Biological Programme 26, Cambridge University Press, Cambridge,
UK.
Parker, T. (199 1). Pollution indicator species (Editorial). Marine Pollution Bulletin
22, 101.
Pascoe, D. and Cram, P. (1977). The effect of parasitism on the toxicity of
cadmium to the three-spined stickleback, Gasterosteus aculeatus L. Journal of
Fish Biology 10,467472.
Pascoe, D. and Woodworth, I. (1980). The effects of joint stress on sticklebacks.
Zeitschrift fur Parasitenkunde 62, 159-163.
Payne, J.F., Kiceniuk, J.W., Squires, W.R. and Fletcher, G.L. (1978). Pathological
changes in a marine fish after a 6-month exposure to petroleum. Journal of the
Fisheries Research Board of Canada 35, 665-667.
Pearse, J.S. and Timm, R.W. (1971). Juvenile nematodes (Echinocephaluspseudo-
uncinatus) in the gonads of sea urchins (Centrostephanus coronatus) and their
effect on host gametogenesis. Biological Bulletin. Marine Biological Laboratory,
Woods Hole, Mass, 140, 95-103.
Pearson, J.C. (1968). Observations on the morphology and life-cycle of Paucivi-
tellosus fragilis Coil, Reid and Kuntz, 1965 (Trematoda: Bivesiculidae).
Perevozchenko, 1.1. and Davydov, O.N. (1974). DDT and its metabolites in some
cestodes in fishes. Gidrobiologicheskii Zhurnal 10, 86-90 (In Russian).
Pilcher, M.W., Whitfield, P.J. and Riley, J.D. (1989). Seasonal and regional
infestation characteristics of three ectoparasites of whiting. Merlangius
merlangus L.. in the North Sea. Journal of Fish Biology 35.97-1 10.
Platzer, E.G. and Adams, J.R. (1967). The life history of a dranunculoid, Philo-
nema oncorhynchi, in Oncorhynchus nerka. Canadian Journal of Zoology 45,
3143.
Poinar, G.O. Jr and Thomas, G.M. (1976). Occurrence of Ascarophis (Nematoda:
Spiruridea) in Callianassa californiensis Dana and other decapod crustaceans.
Proceedings of the Helminthological Society of Washington 43, 28-33.
Pojmhska, T. (1984a). An analysis of seasonality of incidence and maturation of
some fish parasites with regard to thermal factor. I. General methods. Ergasilus
. sieboldi Nordmann, 1832. Acta Parasitologica Polonica 29, 217-228.
Pojmhska, T. (1984b). An analysis of seasonality of incidence and maturation of
some fish parasites with regard to thermal factor. 11. Caryophyllaeus laticeps
(Pallas, 1781). Acta Parasitologica Polonica 29, 229-239.
P.ojmhska, T. (1984~).An analysis of seasonality of incidence and maturation of
some fish parasites with regard to thermal factor. 111. Bunodera luciopercae
(MUller, 1776). Acta Parasitologica Polonica 29, 3 13-321.
Pojminska, T. (1985a). An analysis of seasonality of incidence and maturation of
some fish parasites with regard to thermal factor. IV. Bucephalus polymorphus
Baer, 1927. Acta Parasitologica Polonica 30, 25-34.
Pojminska, T. (1985b). An analysis of seasonality of incidence and maturation of
some fish parasites with regard to thermal factor. V. Digeneans of the genus
Sphaerostoma Rudolphi, 1809. General conclusion. Acta Parasitologica
Polonica 30, 35-46.
Pojminska, T. and Dzika, E. (1987). Parasites of bream (Abramis brama L.) from
the Lake Goslawskie (Poland) affected by long-term thermal pollution. Acta
Parasitologica Polonica 32, 139-1 6 1.
Pojmhska, T., Grabda-Kazubski, S.L., Kazubski, S.L., Machalska, J. and

Niewadomska, K. (1980). Parasite fauna of five species from the Konin lakes
complex, artificially heated with thermal effluents, and from Goplo lake. Acta
Parasitologica Polonica 27, 3 19-357.
Rehulka, J. (1983). The parasites of some fish species of the polluted coarse zone of
the Ostravice River. Prace Vurh Vodnany 12, 63-68.
Renaud, F., Gabrion, C. and Pasteur, N. (1983). Le complexe Bothriocephalus
scorpii (Mueller, 1776): diffkrenciation par Clectrophortse enzymatique des
esptces parasites du Turbot (Psetta maxima) et de la Barbue (Scophthalmus
rhombus).Compte rendu hebdomadaire des sCances de I'AcadCmie des sciences,
Serie III, 296, 127-129.
Renaud, F. and Gabrion, C. (1984). Polymorphisme enzymatique de populations du
B roupe Bothriocephalus scorpii (Muller, 1776) (Cestoda, Pseudophyllidea).

tude des parasites de divers Ttleosttens des cotes du Finistere. Bulletin de la
SociCte' francaise de Parasitologie 2, 95-98.
Riggs, M.R. and Esch, G.W. (1987). The suprapopulation dynamics of Bothrio-
cephalus acheilognathi in a North Carolina reservoir: abundance, dispersion, and
prevalence. Journal of Parasitology 73, 877-892.
Riggs, M.R., Lemly, A.D. and Esch, G.W. (1987). The growth, biomass and
fecundity of Bothriocephalus acheilognathi in a North Carolina cooling reser-
voir. Journal of Parasitology 73, 893-900.
Robert, F., Boy, V. and Gabrion, C. (1990). Biology of parasite populations:
population dynamics of bothriocephalids (Cestoda - Pseudophyllidea) in tele-
ostean fish. Journal of Fish Biology 37, 327-342.
Ruszkowski, J.S. (1934). Etudes sur le cycle Cvolutif et sur la structure des
Cestodes de mer. 111-e partie. Le cycle Cvolutif de TCtrarhynque Grillotia
erinaceus (Van Beneden 1858). MCmoires de l'dcade'mie polonaise des
sciences et des lettres, SCrie B: Sciences naturelles 6, 1-9.
Sakanari, J. and Moser, M. (1985a). Salinity and temperature effects on the eggs,
coracidia, and procercoids of Lacistorhynchus tenuis (Cestoda: Trypanorhyncha)
and induced mortality in a first intermediate host. Journal of Parasitology 71,
583-587.
Sakanari, J. and Moser, M. (1985b). Infectivity of, and laboratory infection with,
an elasmobranch cestode Lacistorhynchus tenuis (Van Beneden, 1858). Journal
of Parasitology 71, 788-791.
Sakanari, J. and Moser, M. (1989). Complete life cycle of the elasmobranch cestode
Lacistorhynchus dolljiusi Beveridge and Sakanari, 1987 (Trypanorhyncha).
Sakanari, J.A., Moser, M., Reilly, C.A. and Yoshino, T.P. (1984). Effects of sub-
lethal concentrations of zinc and benzene on striped bass, Morone saxatilis
(Walbaum), infected with larval Anisakis nematodes. Journal of Fish Biology
24, 553-563.
Samuel, G. and Bullock, W.L. (1981). Life cycle of Paratenuisentis ambiguus
(Van Cleave, 1921) Bullock and Samuel, 1975 (Acanthocephala: Tenuisentidae).
Sandeman, I.M. and Burt, M.D.B. (1972). Biology of Bothrimonus (= Diplocotyle)
(Pseudophyllidea: Cestoda); ecology, life cycle and evolution; a review and
synthesis. Journal of the Fisheries Research Board of Canada 29, 1381-1395.
Sankurathri, C.S. (1974). Effects of thermal effluent on the population dynamics
of Physa gyrina Say (Mollusca: Gastropoda) and its helminth parasites at
Wabumun Lake, Alberta, PhD thesis, University of Alberta, Edmonton, Alberta.
Sankurathri, C.S. and Holmes, J.C. (1976). Effects of thermal effluents on parasites
and commensals of Physa gyrina Say (Mollusca: Gastropoda) and their interac-
tions at Lake Wabamun, Alberta. Canadian Journal of Zoology 54, 1742-1753.
Schell, S.H. (1973). Rugogaster hydrolagi gen. et sp. n. (Trematoda: Aspido-
bothrea: Rugogastridae fam. n.) from the ratfish, Hydrolagus colliei (Lay and
Bennett, 1839). Journal of Parasitology 59, 803-805.
Schleip, W., Herter, K. and Autrum, H. (1937). Hirudineen. Teil 2. VI. Die
Okologie der Hirudineen. Bronns Klassen u. Ord. Tierreichs Leipzig 1937
Bd.4 Abt.3 Buch 4 Teil 2, 321-496.
Siddall, R., Pike, A.W. and McVicar, A.H. (1993). Parasites of Buccinum undatum
(Mollusca: Prosobranchia) as biological indicators of sewage-sludge dispersal.
Journal of the Marine Biological Association of the United Kingdom 73,
931-948.
Simmons, G.E. (1974). Gyrocotyle: a century-old enigma. In “Symbiosis in the
Sea” (W.B. Vemberg, ed.), pp. 195-218. University of South Carolina Press,
USA.
Sindermann, C.J. (1979). Pollution-associated diseases and abnormalities of fish
and shellfish: a review. Fishery Bulletin 76, 717-749.
Skinner, R.H. (1982). The interrelation of water quality, gill parasites and gill
pathology of some fishes from South Biscayne Bay, Florida. Fishery Bulletin 80,
269-280.
Smith, J.W. (1969). The distribution of one monogenean and two copepod parasites
of whiting, Merlangius merlangus (L.) caught in British waters. Nytt Magasinfor
Zoologi 17, 58-63.
Snieszko, S.F. (1974). The effects of environmental stress on outbreaks of
infectious diseases of fishes. Journal of Fish Biology 6, 197-208.
Solangi, M.A. and Overstreet, R.M. (1982). Histopathological changes in two
estuarine fishes, Menidia beryllina (Cope) and Trinectes maculatus (Bloch and
Schneider), exposed to crude oil and its water-soluble fractions. Journal of Fish
Diseases 5 , 13-35.
Stark, G.T.C. (1965). Diplocotyle (Eucestoda), a parasite of Gammarus zaddachi in
the estuary of the Yorkshire Esk, Britain. Parasitology 55, 415-420.
Steven, G.A. (1947). The British Raiidae. Science Progress 138, 220-236.
Strazhnik, L.V. and Davydov, O.N. (1975). The effects of high temperature on the
. biological activities of some fish cestodes. Parazitologiya 9 , 3 7 4 6 (In Russian).
Stunkard, H.W. (1956). The morphology and life-history of the digenetic trema-
tode, Azygia selago Land, 19 10. Biological Bulletin. Marine Biological
Laboratory, Woods Hole, Mass, 111, 248-268.
Sulgostowska, T. (1988). Changes in the parasite fauna of the flounder Platichthys
Jesus dependent on the degree of pollution of the south-westem Baltic Sea.
Wiadomasci Parazytologiczne 34, 59 1-594.
Sunila, I. (1987). Histopathology of mussels (Mytilus edulis L.) from the
Tvarminne area, the Gulf of Finland (Baltic Sea). Annales Zoologica Fennici
24, 55-69.
Symonds, J.D. (1972). Infestation of Nephrops norvegicus (L.) by Stichocotyle
nephropis Cunningham in British waters. Journal of Natural History 6,423426.
Thoney, D.A. and Burreson, E.M. (1987). Morphology and development of the
adult and cotylocidium of Multicalyx cristata (Aspidocotylea), a gall bladder
parasite of elasmobranchs. Proceedings of the Helminthological Society of
Washington 54, 96-104.
Thulin, J. (1984). The impact of some environmental changes on the parasite
fauna of cod in Swedish waters (Abstract). Abstracts of the Fourth European

Multicolloquium of Parasitology, 239-240.
Thulin, J. (ed.) (1986). Report of the ICES workshop on the use of pathology in
studies of the effects of contaminants. International Council for the Exploration
of the Sea CM 1986E: 40, Ref. H, 64 pp.
Thulin, J. (1987). Some diseases and parasites of trout (Salmo trutta) attracted to a
hot water effluent. In “Parasites and Diseases in Natural Waters and Aquaculture
in Nordic Countries” (A. Stenmark and G. Malmberg, eds), p. 56. Zoo-tax,
Stockholm, Sweden.
Thulin, J. Hoglund, J. and Lindesjoo, E. (1988). Diseases and parasites of fish in a
bleached kraft mill effluent. Water Science Technology 20, 179-180.
Turner, H.M. (1986). Prevalence of larval Proctoeces maculata (Trematoda:
Fellodistomidae) infection in hooked mussels from a Louisiana estuary.
Proceedings of the Helminthological Society of Washington 53, 130-13 1.
Uhazy, L.S. (1977a). Development of Philometroides huronensis (Nematoda:
Dranunculoidea) in the intermediate and definitive hosts. Canadian Journal of
Zoology ,55, 265-273.
Uhazy, L.S. (1977b). Biology of Philometroides huronensis (Nematoda: Dranun-
culoidea) in the white sucker (Catostomus commersoni). Canadian Journal of
Zoology 55, 1430-1441.
Uspenskaya, A.V. (1960). Parasitofaune des crustaces benthiques de la mer de
Barents (Expos6 prtliminaire). Annales de Parasitologie Humaine et Comparte
35,221-242.
Valtonen, E.T. and Koskivaara, M. (1987). The effect of environmental stress on
trematodes of perch and roach in central Finland (Abstract). In “Actual
Problems in Fish Parasitology”, p. 103, 2nd International Symposium of
Ichthyoparasitology, Tihany, Hungary.
Valtonen, E.T. and Koskivaara, M. (1989). Effects of effluent from a paper and
pulp mill on parasites of the roach in central Finland. Soviet-Finnish Symposium
on Fish Parasites of North-Westem Europe, Petrozavodsk, USSR, 163-168.
Valtonen, E.T. and Taskinen, J. (1988). Rhipidocotyle campanula in its first and
second intermediate hosts in central Finland; associated with pollution?
(Abstract). Abstracts of the Vth European Multicolloquium of Parasitology,
Budapest, Hungary, 110.
Valtonen, E.T., Koskivaara, M. and Brummer-Korvenkontio, H. (1987). Parasites
of fishes in central Finland in relation to environmental stress. Biological
Research Report, University of Jyvaeskylae 10, 129-130.
Vladimirov, V.L. and Flerov, B.A. (1975). Susceptibility to ichthyophthiriasis in
fish following exposure to phenol and polychloropinene poisoning. Biologiya
Vnutrennikh Vod Informatsionnyi Byulleten, 25, 35-37 (In Russian).
Walkey, M. and Karting, W. (1985). Thermal alterations of pymvate kinases in the
fish tapeworm Bothriocephalus acheilognathi Yamaguti, 1934. Zeitschrift fur
Parasitenkunde. 71, 527-532.
Williams, H.H. (1965). Observations on the occurrence of Dictyocotyle coeliaca
and Calicotyle kroyeri (Trematoda: Monogenea). Parasitology, 55, 20 1-207.
Williams, H.H. (1965). The ecology, functional morphology and taxonomy of
Echeneibothrium Beneden, 1849 (Cestoda: Tetraphyllidea), a revision of the
genus and comments on Discobothrium Beneden, 1870, Pseudanthobothrium
Baer, 1956, and Phormobothrium Alexander, 1963. Parasitology, 56, 227-285.
Williams, H.H., Colin, J.A. and Halvorsen, 0. (1987). Biology of gyrocotylideans
with emphasis on reproduction, population ecology and phylogeny. Parasitology

95, 173-207.
Williams, H.H. and Halvorsen, 0. (1971). The incidence and degree of infection
of Gadus morhua L., 1958 with Abothrium gadi Beneden, 1871 (Cestoda:
Pseudophyllidea). Norwegian Journal of Zoology 19, 193-199.
Winch, J. (1983). The biology of Atrispinum labracis n. comb. (Monogenea) on the
gills of the bass, Dicentrarchus labrax. Journal of the Marine Biological
Association of the United Kingdom 63, 915-927.
Winstead, J.T. and Couch, J.A. (1988). Enhancement of protozoan pathogen
Perkinsus marinus infections in American oysters Crassostrea virginica
exposed to the chemical carcinogen nitrosodiethylamine (DENA). Diseases of
Aquatic Organisms 5, 205-213.
Wojdani, A. and Alfred, L.J. (1984). Alterations in cell-mediated immune
functions induced in mouse splenic lymphocytes by polycyclic aromatic hydro-
carbons. Cancer Research 44, 942-945.
Wright, C.A. (1956). Studies on the life-history and ecology of the trematode genus
Renicola Cohn, 1904. Proceedings of the Zoological Society of London 126,
149.
Xylander, W.E.R. (1986). Zur Biologie und Ultrastntktur der Gyrocotylida und
Amphilinida sowie ihre Stellung im phylogenetischen System der Platyhel-
minthes. PhD dissertation of the Georg-August-Universitat of Gottingen. Dissera-
tion zur Erlangung des Doktorgrades der Mathematisch-Naturwissenschaftlichen
Fachbereiche der Georg-August-Universitat zu Gottingen.
Xylander, W.E.R. (1989). Investigations on the biology of Gyrocotyle urna
(Cestoda) and considerations on its life cycle. Verhandlungen Deutschen
Zoologischen Gesellschafi 82, 25 1 (In German).
Yevich, P.P. and Barszcz, C.A. (1983). Histopathology as a monitor for marine
pollution. Results of the histopathologic examination of the animals collected for
the US 1976 Mussel Watch Program. Rapports et proces-verbaux des Rtunions.
Conseil Permanent International pour l’exploration de la Mer 182, 96-102.
Young, R.T. (1954). A note on the life cycle of Lacistorhynchus tenuis
(Van Beneden, 1858), a cestode of the leopard shark. Proceedings of the
Helminthological Society of Washington 21, 112.
Zeeman, M.G. and Brindley, W.A. (1981). Effects of toxic agents upon fish
jmmune systems: a review. In “Immunologic Considerations in Toxicology,
Volume 11” (R.P. Sharma, ed.), pp. 1-60. CRC Press, Boca Raton, Florida.
Variation in Echinococcus: Towards a
Taxonomic Revision of the Genus
'
. R.C.A. Thompson. A.J. Lymbery2and C.C. Constantine'
WHO Collaborating Centre for the Molecular Epidemiology of Parasitic

Infections and Institute for Molecular Genetics and Animal Disease.
'School'of Veterinary Studies. Murdoch University. Western Australia.
6150. Australia and Western Australian Department of Agriculture.
Bunbury. Western Australia. 6230. Australia
1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 146
1.1. Status of hydatid disease ...................... 146
1.2. Variation in Echinococcusand control of hydatid disease . . . .
.. 146
1.3. Taxonomic considerations ..................... 147
.
2 Species Concepts and their Application ................. 150
2.1. Definition of a species ....................... 150
.........
2.2. An evolutionary species concept for Echinococcus 151
.
3 Identification of OTUs . . . . . . . . . . . . . . . . . . . . . . . . . . 152
3.1. Echinococcus granulosus . . . . . . . . . . . . . . . . . . . . . . 153
3.2. Echinococcus multilocularis . . . . . . . . . . . . . . . . . . . . . 158
. ...............
3.3. Echinococcus vogeli and E oligarthrus 159
.
4 Phylogeny of OTUs ........................... 159
. ...................
5 Delimitation of Evolutionary Species 162
5.1. Species 1 (Echinococcussp.) .................... 162
.................
5.2. Species 2 (Echinococcusortleppin 164
.................
5.3. Species 3 (Echinococcusequinusn 165
...............
5.4 Species 4 (Echinococcusmultilocularis) 165
. ......
5.5 Species 5 and 6 (Echinococcus vogeli and E oligarthrus) 166
................
5.6. Species 7 (Echinococcusgranulosus) 166
6. Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 167
Acknowledgements ........................... 168
References ............................... 168
ADVANCES IN PARASITOLOGY VOL 35 Copyrighr Q 1995 Academic Press Lim'red

ISBN 0-12-031735-4 All righfs of reproduction in any form reserved
146 R.C.A. THOMPSON ET AL
1. INTRODUCTION
1.1. Status of Hydatid Disease
Hydatid disease (echinococcosis) is one of the most important parasitic

zoonoses and remains a public health and economic problem of global
proportions. Surgery is the only effective treatment in humans, although
the prognosis with the multivesicular (alveolar) form of the disease caused
by E . multilocularis is often poor. Control efforts have had little impact on
the worldwide status of hydatid disease, :with human activity the major
factor in perpetuating transmission. New foci of infection and regions of
endemicity have recently been recognized and there is increasing evidence
of the causative agents extending their range into areas previously con-
sidered to have been free of infection (Thompson and Allsopp, 1988;
Schantz, 1991; Ballek, 1991; Brochier et al., 1992). In addition, interac-
tion between wild and domestic cycles of transmission is an emerging
cause for concern (Eckert and Thompson, 1988; Hildreth et al., 1991;
Schantz, 1991; Thompson, 1992; Storandt and Kazacos, 1993).
1.2. Variation in Echinococcus and Control of Hydatid Disease
There are currently four recognized species in the genus Echinococcus: E .

granulosus, E . multilocularis, E . oligarthrus and E. vogeli. In the most
well-studied species, E. granulosus and E. multilocularis, a large number
of intraspecific variants, designated formally as subspecies or informally as
strains, have been described (Thompson and Lymbery, 1988, 1990a, b;
Table 1).
Because of the extensive variation in Echinococcus, it is very important
to characterize the aetiological agents in different endemic areas in order to
determine transmission patterns, particularly where there is the possibility
of interaction between cycles (Thompson, 1994). For example, in Britain,
where there are two cycles of transmission, interaction is unlikely since
each cycle is associated with the perpetuation of a distinct strain exhibiting
preference for a different species of intermediate host (Thompson and
Smyth, 1975; Thompson, 1991). In contrast, on the mainland of Australia,
although E . granulosus is maintained in different cycles of transmission
involving either domestic or wild host assemblages, there is no evidence of
genetic distinctness between the parasites found in these cycles (Lymbery
et al., 1990; Thompson and Lymbery, 1990a; Hope et al., 1991). This is
significant to the control of hydatid disease in Australia since wild and
domestic cycles of transmission may interact in areas where they overlap
VARIATION IN ECHINOCOCCUS TOWARDS A TAXONOMIC REVISION OF THE GENUS 147
(Thompson, 1992; Constantine et af., 1993). Parasite characterization and

the demonstration of strain differences is particularly important in regions
where there is more than one species of intermediate host and therefore the
possibility of different cycles of transmission and sources of infection for
humans. This is the case in the Middle East, Africa and China where
numerous intermediate host species harbour cysts of E. grunulosus
(Thompson and Lymbery, 1988; Thompson, 1994).
Developmental differences between strains of Echinococcus, such as
variation in the onset of egg production, will affect transmission and
impede control efforts when regular, adult cestocidal treatment is used to
break the cycle (Kumaratilake et af., 1983; Thompson et a f . , 1984; Eckert
et al., 1989). Accurate surveillance of hydatid disease in humans and the
future development of immunoprophylactic strategies may be jeopardized
by demonstrated antigenic differences between isolates of Echinococcus
(Cameron, 1960; Huldt et al., 1973; Gottstein et af., 1983; Lightowlers et
af., 1984; Gottstein, 1991; Lightowlers and Gottstein, 1994).
Strains of varying pathogenicity will influence the prognosis in patients
with hydatid disease. This is of particular significance in cases of alveolar
hydatid disease (Kawase and Yagi, 1985; Liance et al., 1990), but is also
important for cystic hydatid disease, caused by E. granufosus.For example,
the sylvatic strain of E. granulosus in northern North America causes a
benign infection in humans (Wilson et af., 1968) whereas, in parts of
Kenya and Libya, it has been suggested that there are local virulent strains
of E. granulosus (French et al., 1982; Gebreel et al., 1983). There is also
evidence that certain strains of E. granulosus, such as those adapted to
horses and pigs, may not be infective to humans (Thompson and Lymbery,
1988, 1991; Shablovskaya et af., 1989).
It has been suggested that strains of Echinococcus may differ in their
response to particular chemotherapeutic regimes (Saimot et af., 1981;
Schantz et af., 1982; Kammerer and Schantz, 1984). This is supported by
detailed information being obtained on the biochemical differences between
strains, which is vital for the development of new chemotherapeutic agents
(McManus and Smyth, 1982; McManus and Bryant, 1994).
1.3. Taxonomic Considerations
In a review of the nature, extent and significance of variation within the

genus Echinococcus in 1988, we enumerated the described species, sub-
species and strains of Echinococcus and attempted to provide a workable
definition for the nebulous term “strain” (Thompson and Lymbery, 1988).
We anticipated that the application of molecular characterization tech-
niques would provide additional data on genetic variation within and
Table 1 Species and strains of Echinococcus, according to current classification.
Current species Strain OTU Known intermediate hosts Known definitive Probableb geographic distribution
designation Codea hosts
~
E . granulosus Sheep strain GSH Sheep, cattle, pigs, camels, Dog, fox, dingo, Australian mainland, Europe, USA,
goats, macropods, humans jackal, hyena New Zealand, Africa, China, Middle
East, South America, Russia
E. granulosus Tasmanian sheep GTA Sheep, cattle? humans Dog (fox) Tasmania
Strain
E. granulosus Buffalo strain(?) GBU Buffalo (cattle?) (humans?) Dog (fox?) Asia
E. granulosus Horse strain GHO Horses and other equines Dog Europe, Middle East, South Africa
(New Zealand? USA?)
E. granulosus Cattle strain GCT Cattle, humans Dog Europe, South Africa, India, Sri
Lanka, Russia
E. granulosus Camel strain GCM Camels, goats, cattle? Dog Middle East, Africa
humans?
E. granulosus Pig strain GPI Pigs, humans? Dog Europe, Russia, South America
E. granulosus Cervid strain' ? Cervids, humans Wolf, dog North America, Eurasia
E. granulosus Lion strain' ? Zebra, wildebeest, warthog, Lion Africa
bushpig, buffalo, various
antelope, giraffe?
hippopotamus? (humans?)
E. rnultilocularis European strain' MEU Rodents, humans Fox, dog, cat Europe, China(?)
E. multilocularis Alaskan strain' ? Rodents, humans Fox, dog, cat Alaska
E. rnultilocularis North American h4NO Rodents, humans Fox, dog, cat North America
Strain'(?)
E. rnultilocularis Hokkaido strain'(?) ? Rodents, pig, horse, Fox, dog, cat Japan
humans
E . vogeli None reported VOG Rodents, humans Bush Dog South America
E. oligarthrus None reported OLI Rodents, humans Felids South America
a Operational Taxonomic Unit (see text, section 2.2).

With some strains, the geographic range of isolates which have been characterized simultaneously using morphological and genetic
criteria is limited (see text).
No detailed genetic characterization; at present separated on the basis of morphological, biological and epidemiological features.
between populations of Echinococcus and thus allow a more reliable

foundation for determining genetic relationships and for establishing an
acceptable and realistic classification for the genus. If anything, we under-
estimated the impact of molecular genetic studies. The last six years have
provided a wealth of new data that do not fit comfortably with the current
subgeneric classification (Lymbery, 1992; Bowles and McManus, 1993a).
In this review, we outline how we believe a taxonomic revision of the
genus should proceed.
2. SPECIES CONCEPTS AND THEIR APPLICATION
2.1. Definition of a Species
The first step in a taxonomic revision of the genus Echinococcus is to

establish an appropriate species concept. The most widely used definition
of a species is the biological species concept of Mayr (1942): “Species are
groups of actually or potentially interbreeding natural populations, which
are reproductively isolated from other such groups”. An obvious deficiency
of the biological species concept is that it is applicable only to sexually
reproducing, outcrossing organisms. Kumaratilake and Thompson (1982)
suggested that the mode of reproduction in Echinococcus rendered this
concept inappropriate for the genus. Species of Echinococcus have a
sexually reproducing phase every generation, however, and although
exclusive self-fertilization is often assumed (Smyth and Smyth, 1969;
Kumaratilake et al., 1979; Nadler, 1987), population genetic data indicate
that outcrossing does occur, at least in some populations (Lymbery, 1992,
1994).
Unfortunately, there is no definitive answer to the question of how rare
outcrossing needs to be to prevent meaningful application of the biological
species concept (Templeton, 1989). In any case, there are other deficiencies
of the concept which make it desirable to search for alternatives. Specifi-
cally, the biological species concept has been criticized because it is
difficult to apply to allopatric populations, because it emphasizes reproduc-
tive isolating mechanisms at the expense of mate recognition mechanisms,
because it overstates the unifying power of gene flow, and because it
ignores historical relationships between populations (Mishler and
Donoghue, 1982; McKitrick and Zink, 1988; Templeton, 1989).
In general, two different types of species concepts have been proposed in
response to the perceived shortcomings of the biological species concept.
Phylogenetic species concepts give primacy to the pattern of relationships
VARIATION IN ECHINOCOCCUS: TOWARDS A TAXONOMIC REVISION OF THE GENUS 151
between organisms, rather than to the processes responsible for their

differentiation, whereas evolutionary species concepts consider processes
additional to interbreeding (and its converse, reproductive isolation) as
equally important in providing evolutionary cohesiveness to species. A
general solution to the species problem is probably not achievable,
because all species concepts which consider species to be real evolu-
tionary units must make reference, implicitly or explicitly, to future events
(O’Hara, 1993). Our preference for evolutionary, over phylogenetic con-
cepts has been discussed elsewhere (Lymbery, 1994). In this section, we
wish to amplify the evolutionary species concept we favour and provide
operational rules for its implementation in the genus Echinococcus.
2.2. An Evolutionary Species Concept for Echinococcus
We take as our starting point the definition of Wiley (1978): “A species is a

single lineage of ancestral descendant populations of organisms which
maintains its identity from other such lineages and which has its own
evolutionary tendencies and historical fate”. There are two parts to this
definition:
1. A species is a single lineage of ancestral descendant populations.
Evolutionary species must be consistent with inferred historical rela-
tionships between their constituent populations (where populations are
defined as geographically localized units of organisms united by com-
mon descent [asexual species] or recombination [sexual species]).
Species, in essence, are monophyletic groups of populations. Evo-
lutionary species differ from supraspecific taxa in that these taxa,
even if monophyletic, cannot be regarded as a single lineage; they are
instead separate lineages grouped by past history. Wiley (1981) views
evolutionary species as the largest evolving phylogenetic entities.
2. which maintains its identify from other such lineages and which has its
own evolutionary tendencies and historical fate. To be considered part
of the same evolutionary species, populations or sublineages must be on
the same evolutionary trajectory. This is the aspect of Wiley’s definition
which has proved operationally most intractable; how can we determine
whether populations are or are not independently evolving units? We
believe that the best answer to this question comes from considering
the population genetic processes responsible for maintaining pheno-
typic and genetic cohesion within a species (Templeton, 1989). From
this mechanistic viewpoint, an evolutionary species is defined by the
boundaries of action of the forces of gene flow, genetic drift and natural
selection. That is, species must possess either genetic exchangeability
(new genetic variants can replace old variants within the species
through gene flow) or demographic exchangeability (new genetic
variants can replace old variants within the species through genetic
drift and natural selection). This definition recognizes the importance
of interbreeding, but it becomes just one of a class of mechanisms
responsible for maintaining cohesion and the evolutionary integrity of
a lineage.
Determining species status is a two-step procedure; organisms must first
be grouped into taxa and then these taxa ranked into their appropriate
category (Donoghue, 1985). We will take the two parts of the evolutionary
species concept described above as our grouping and ranking criteria for
Echinococcus. The operational steps for delimiting species within the
genus are then as follows:
1 . Identify basal taxa (operational taxonomic units [OTUs]; Sneath and
Sokal, 1973). Ideally these will be geographically localized populations,
but sampling limitations will often necessitate the use of more inclusive
sublineages.
2. Reconstruct the phylogeny of OTUs. This will provide a hierarchy of
monophyletic groups suitable for ranking into their appropriate category.
3. Rank as evolutionary species those most inclusive groups having the
potential for genetic or demographic exchangeability:
(a) OTUs which are sympatric and yet maintain genetic distinctness
clearly do not possess exchangeability and should be considered
separate species;
(b) The exchangeability of allopatric OTUs must be inferred from
information on their genetic and ecological similarity. The appro-
priateness of these criteria for inferring exchangeability is discussed
by Lymbery (1994). They do not provide an infallible guide. Each
case must be considered separately and a decision on species status
made according to all the information available at that time. It is
inevitable that this procedure will produce errors, but the delimita-
tion of evolutionary species should be considered an hypothesis to
be tested by further data.
3. IDENTIFICATION OF OTUs
We will take as our basal taxa, not populations, but those groups of
organisms which have already been defined as different species, sub-
species or strains of Echinococcus (Table 1). The reasons justifying their
classification and categorization have previously been discussed in detail
(Rausch, 1967, 1986; Kumaratilake and Thompson, 1982; Thompson and

Lymbery, 1988,1990a). However, in the six years since this subject was last
reviewed (Thompson and Lymbery, 1988), more information has become
available, particularly with the application of molecular characterization
techniques. Such studies have involved restriction fragment length poly-
morphism (RFLP) analysis of ribosomal DNA (rDNA) or other genomic
regions (McManus and Rishi, 1989; Thompson and Lymbery, 1991; Vogel
et al., 1991; Bowles and McManus, 1993b; Eckert et al., 1993), and
sequencing of the mitochondrial cytochrome c oxidase subunit I (COI)
and NADH dehydrogenase 1 (ND1) genes (Bowles et al., 1992a; Bowles
and McManus, 1993~).These DNA regions appear to be variable enough to
differentiate most biologically distinct groups, yet conservative enough not
to exhibit extensive within-population polymorphism; they are therefore
ideal for the identification of basal OTUs. In addition, there have been a
number of comparative studies that have provided a clearer picture of some
forms of Echinococcus that previously were of uncertain status. Our
purpose here is therefore to update what we know about the nature and
extent of variation in Echinococcus in order to provide a sound basis for a
phylogenetic analysis. For convenience, we will for now retain the standard
nomenclature shown in Table 1. As our analysis proceeds, however, it will
be clear that the current classification of these OTUs may not conform to
biological reality.
3.1. Echinococcus granulosus
3.1.1. Sheep Strain

The widespread distribution and apparent uniformity of the sheep strain
of E. granulosus has been emphasized by a number of recent studies.
Molecular characterization of rDNA and of mitochondrial COI and NDl
genes has confirmed the genetic uniformity of the sheep strain from a range
of endemic areas (McManus and Rishi, 1989; Bowles et al., 1992a; Bowles
and McManus, 1993b, c). These studies have also confirmed previous
observations that a range of other mammalian species may serve as inter-
mediate hosts. In addition, the sheep strain has been shown to be locally
distinct from other forms of E. granulosus in many endemic areas, including
Tibet (Hu et al., 1993), China, Lebanon, Turkey, Italy (Bowles and
McManus, 1991, 1993a), Spain (Siles-Lucas et al., 1993), Jordan (Said
et al., 1988; Bowles and McManus, 1991; Hijjawi et al., 1992), Russia
(Yastreb, 1986; Skvortsova and Artemenko, 1987), India (Singh et al.,
1988), United Kingdom (Clarkson and Walters, 1991), Kenya (Wachira
et al., 1993) (and see Thompson and Lymbery, 1988 for earlier studies).
3.1.2. Tasmanian Sheep Strain

Kumaratilake et al. (1983) and Kumaratilake and Thompson (1983, 1984a,
b) found morphological, biochemical and developmental differences
between populations of E. granulosus from sheep in Tasmania and on the
mainland of Australia. This phenotypic variation has now been substantiated
by the demonstration of small, yet significant, genetic differences (Lymbery
and Thompson, 1988; Bowles et al., 1992a; Bowles and McManus, 1993~).
The status of a genetically distinct population of E . granulosus found
in cattle on King Island, north of Tasmania, requires further study
(Constantine et al., 1991). The origin of this new focus of infection is
not known and there is little information on the phenotypic characteristics
df the parasite or the epidemiology of hydatid disease on the island.
3.1.3. Buffalo Strain ( ?)

The buffalo is the most significant host for maintaining the life cycle of E .
granulosus in Asia (Gill and Venkateswara, 1967; Islam, 1982; Irshadullah
et al., 1989). Previous studies have shown that E . granulosus from buf-
faloes differs morphologically and developmentally from that in sheep and,
based on published data, Thompson and Lymbery (1988) suggested that the
form in buffaloes may be the same as the cattle strain of E. granulosus.
Molecular genetic data have confirmed the occurrence of the cattle strain in
buffaloes from India, but have also revealed that buffaloes may harbour the
sheep strain and a unique genotype not seen in other intermediate host
species (Bowles et al., 1992a; Bowles and McManus, 1993a, b, c). This
form, which we now tentatively designate the buffalo strain, is genetically
very similar to the sheep strain. There is very little information on the
transmission patterns of Echinococcus in India and the extent of strain
variation (Singh et al., 1988), and clearly more research is required to
determine whether there is a distinct form of E . granulosus adapted to
buffaloes.
3.1.4. Horse Strain

As with the sheep strain of Echinococcus, the characteristics of the horse
strain are now well defined (Thompson and Lymbery, 1988, 1991;
McManus et al., 1989; Smyth, 1990). It represents a genetically (McManus
and Rishi, 1989; Bowles et al., 1992a; Bowles and McManus, 1993b,c) and
morphologically (Kumaratilake et al., 1986) uniform strain adapted to
horses and other equines and thus differs from the sheep strain in having
a much more limited intermediate host range (Table 1). The horse strain is
VARIATION IN ECHINOCOCCUS TOWARDS A TAXONOMIC REVISION OF THE GENUS 155
geographically widely distributed (Kumaratilake et al., 1986; Hijjawi et al.,

1992) and appears to be non-infective to humans.
3.1.5. Cattle Strain

A form of E. granulosus adapted to cattle as intermediate host was initially
suggested by Verster (1965) and later characterized by Thompson et al.
(1984), who demonstrated the distinctness of E. granulosus adapted to
cattle with respect to the morphology and precocious development of
the adult worm. The genetic distinctness of the cattle strain has been
confirmed by a number of studies (McManus and Rishi, 1989; Thompson
and Lymbery, 1991; Bowles et al., 1992a; Bowles and McManus, 1993b, c;
Eckert et al., 1993). This strain of E. granulosus has now been shown to be
widely distributed throughout Europe, including Germany (Hahn et al.,
1988; Worbes et al., 1989; Janssen et al., 1990), Holland (Bowles and
McManus, 1991, 1993a), Belgium (De Rycke, 1968; Janssen et al., 1990),
Ireland (Harrison et al., 1986), Denmark (Jorgensen and Hansen, 1984),
Russia (Skvortsova and Artemenko, 1987; Yastreb and Skvartsova, 1991),
South Africa (Verster, 1965), India (Abraham et al., 1980; Sanyal and Sinha,
1983; Bowles and McManus, 1991, 1993a) and Sri Lanka (Dissanake,
1957). Epidemiological and molecular data also suggest that the cattle
strain is infective to humans (Thompson, 1988; Janssen et al., 1990;
Bowles et al., 1992b).
3.1.6. Camel Strain

The important role of the camel as intermediate host of E. granulosus has
long been recognized in different parts of the world, particularly the
Middle East and Africa (reviewed in Eckert et al., 1989). E . granulosus
in camels has recently been shown to be quite distinct in its morphology
and development (Said et al., 1988; Eckert et al., 1989) and molecular
characterization of camel isolates from the Middle East (Thompson and
Lymbery, 1991; Eckert et al., 1993) and Kenya, Sudan and Somalia
(McManus and Rishi, 1989; Bowles et al., 1992a; Bowles and McManus
1993b, c; Wachira et al., 1993) have confirmed their genetic distinctness.
There are some discrepancies, however, between data from the Middle East
and Africa. Epidemiological evidence from several areas in the Middle
East has long suggested that camels are an important reservoir for infec-
tions in people (Eckert et al., 1989). In addition, isolates from camels in
Egypt and Jordan are morphologically and genetically distinct from strains
in sheep and cattle (Said el al., 1988; Eckert et al., 1989, 1993; Thompson
and Lymbery, 1991; Morgan et al., submitted for publication). In Kenya, it
has been emphasized that isolates of E. granulosus from different species
of intermediate hosts are uniform morphologically, despite their genetic

differences (Bowles and McManus, 1993a; Wachira et al., 1993). More-
over, the majority of camel isolates examined from Kenya could be
differentiated from the sheep strain based on molecular criteria but
showed no affinity with isolates of human origin. It does not seem likely
that the strain adapted to camels in Kenya is different from that in camels
in the Middle East, particularly since there is evidence that E. granulosus
was introduced into Africa from the Middle East (Nozais, 1987). It is also
interesting that in Somalia where camels and dogs are frequently infected
with E. granulosus, human cases of hydatid disease have not been
recorded, yet primates are susceptible to experimental infection following
ingestion of eggs of Somalian camel/dog origin (Macchioni et al., 1987).
More work is clearly required on the epidemiology of hydatid disease in
these regions. In addition, we need a direct genetic comparison of isolates
from camels in the Middle East and Africa and a reappraisal of the
morphological characteristics of E. granulosus of sheep and camel origin
in Kenya.
3.1.7. Pig Strain

Pigs serve as an important intermediate host for E. granulosus in several
areas including eastern Europe and the former Soviet Union (for references
see Thompson and Lymbery, 1988; Eckert et al., 1993) and, more recently,
Mexico (Cruz-Reyes et al., 1990). Comparative studies have demonstrated
that the forms of E. granulosus adapted to pigs share a series of distinct
morphological, developmental and epidemiological characteristics which
serve to separate the pig strain from forms of E. granulosus utilizing other
intermediate hosts (Yastreb 1986; Skvortsova and Artemenko, 1987;
Shqbovskaya et al., 1989; Thompson and Lymbery, 1991; Eckert et al.,
1993). Genetic differences between isolates from pigs and other host
species have been demonstrated using DNA characterization (McManus
and Rishi, 1989; Thompson and Lymbery, 1991; Bowles et al., 1992;
Bowles and McManus, 1993b, c; Eckert et af., 1993), and genetic similarity
has been found between pig isolates from widely separated geographical
areas, Mexico and Poland, using random amplified polymorphic DNA
(RAPD) techniques (Morgan et al., submitted for publication).
3.1.8. Cervid Strain (?)

Echinococcus granulosus is found in large deer such as elk (moose) and
reindeer, in northern North America and Eurasia (Rausch, 1986, 1994). As
detailed previously (Thompson and Lymbery, 1988), there are considerable
epidemiological and phenotypic features which serve to separate the cervid
form of E. grunulosus from other strains. The genetic characteristics of

Echinococcus affecting cervids, however, have not yet been clearly
defined. In a recent study in which an isolate of E. grunulosus from a
north American moose was compared with other strains of Echinococcus
using RAPD analysis, the cervid isolate was found to be distinct but with
similafities to camel and cattle strains (Morgan et al., submitted for pub-
lication). This supports previous morphological and biochemical studies
(Thompson and Lymbery, 1988), but further genetic studies are needed.
3.1.9. Lion Strain (?)

One of the most interesting yet least studied forms of E. granulosus is that
which utilizes the lion as definitive host in several African countries
(Macpherson, 1986; Thompson and Lymbery, 1988). The unusual (for
E. grunulosus) utilization of a felid as definitive host, morphological
differences (Ortlepp, 1937; Graber and Thal, 1980) and the existence
of sylvatic cycles involving warthog, zebra, wildebeest, buffalo and
antelope, support the suggestion by Macpherson (1986) and Rausch
(1986, 1994), that Ortlepp’s (1937) original specific ranking for this
form of Echinococcus as E. felidis is correct. However, as for the cervid
form of Echinococcus, genetic characterization of isolates from lions is
essential to determine taxonomic affinities with other species and strains of
Echinococcus.
3.1.10. Other Strains

In a previous review (Thompson and Lymbery, 1988), it was suggested that
phenotypic differences between E. grunulosus from wild and domestic
hosts on the mainland of Australia provided evidence for the occurrence
of a distinct strain in macropod marsupials, with dingoes as the principal
definitive host. This has not been confirmed by subsequent genetic studies.
Allozyme and RFLP data do not indicate significant genetic differences
between isolates of Echinococcus from macropods and sheep (Lymbery et
ul., 1990; Hope et ul., 1991; Bowles and McManus, 1993b), and the
phenotypic differences previously reported appear to be host induced
(Hobbs et ul., 1990).
Similarly, Pandey (1972) proposed a separate strain of E. grunulosus
infecting goats, but molecular characterization of isolates from Africa
and China (McManus and Rishi, 1989; Bowles et ul., 1992a; Bowles and
McManus, 1993a, b) have shown only that goats harbour both sheep and
camel strains of E. grunulosus.
European hares have been found to be commonly infected with Echino-
coccus in Argentina, and Thompson and Lymbery (1988) suggested that
the existence of a distinct lagomorph strain warranted further investigation.

We are not aware of any further studies on this question.
3.2. Echinococcus multilocularis
Compared to E. granulosus, the nature and extent of variation within E.

multilocularis has been little studied. Although there is evidence of varia-
tion in morphology, pathogenesis, antigenic characteristics, development
and host specificity (Eckert and Thompson, 1988; Thompson and Lymbery,
1988; Gottstein, 199l), information on genetic diversity between isolates
of E. multilocularis is limited.
Rausch (1967) recognized three subspecies of E. multilocularis; E. multi-
locularis multilocularis in Eurasia, E. rnultilocularis sibiricensis in North
America and E. multilocularis kazakhensis in Kazakhstan. The existence of
E. m. kazakhensis appears doubtful (Thompson and Lymbery, 1988). There
is evidence of both morphological and biological differences between
European, Alaskan and other North American populations of E. multi-
locularis (Eckert and Thompson, 1988; Bartel et al., 1992; Table 1).
Distinct biological characteristics have also been reported for E. multi-
locularis in Hokkaido, Japan. Unlike other populations of E. multilocularis
(Pfister and Frank, 1988), it is infective to pigs and natural infections have
been frequently recorded in Hokkaido (Kamiya, 1988). In addition, recent
reports suggest that, atypically, Norway rats are susceptible to infection
with the form of E. multilocularis in Hokkaido (Okamoto et al., 1992).
It is only recently that isolates of E. multilocularis from different
endemic areas have been studied genetically. Vogel et al. (1991) com-
pared ten isolates (seven Swiss, one French, one German, and one
Canadian) by Southern blotting using the Echinococcus-specific genomic
probe PAL 1, and demonstrated heterogeneity between isolates. Differences
were found between Swiss isolates as well as between isolates from
Switzerland and other endemic areas, but no attempt was made to correlate
genetic variability with phenotypic characteristics. Bowles et al. (1992a)
and Bowles and McManus (1993~)found differences in mitochondria1 COI
and ND1 sequences between isolates of E. mulrilocularis from Europe and
from North America and China. This conflicts with the traditional separa-
tion of Eurasian and North American subspecies or strains. More recently,
the randomly amplified polymorphic DNA technique was applied to a
range of E. multilocularis isolates and revealed homogeneity among
European isolates and a Japanese isolate of E. multilocularis from
Hokkaido (Morgan et al., submitted for publication). In contrast, an
Alaskan isolate of the parasite exhibited quite distinct RAPD profiles
clearly separating it from other forms of E . multilocularis (Morgan et

al., submitted for publication).
Clearly, more research is required on the genetic characterization of E .
multilocularis populations in different endemic areas. This is particularly
important since E . multilocularis appears to be extending its range in many
areas, for example, in central North America (Storandt and Kazacos, 1993)
and parts of Europe (Brochier et al., 1992) and the biological heterogeneity
which has been demonstrated between isolates (e.g. Bartel et al., 1992)
should be complemented by molecular genetic studies. At this stage, we
will recognize as OTUs only the two populations shown to be genetically
different by Bowles et al. (1992a) and Bowles and McManus (1993~).
3.3. Echinococcus vogeli and Echinococcus oligarthrus
Bowles et al. (1992a) and Bowles and McManus (1993b, c) found these
species to be quite distinct genetically from each other and from all other
species and strains of Echinococcus examined. Only two isolates of E.
vogeli and one isolate of E. oligarthrus were sequenced, however, and we
are aware of no other studies which have examined genetic, morphological
or biological variation within these species.
4. PHYLOGENY OF OTUs
There has been little previous work on the phylogeny of species and strains
of Echinococcus. Lymbery ( 1992) analysed published morphological data
for a number of strains of E . granulosus and E . multilocularis, but the
accuracy (sensu Hillis and Bull, 1993) of the resultant phylogeny was
severely constrained by the availability and quality of morphological
characters.
The DNA sequence data published by Bowles et al. (1992a) and Bowles
and McManus (1993~)provide much more reliable characters with which
to reconstruct the phylogeny of OTUs of Echinococcus. Available for
analysis are the published sequences of a 366 bp fragment of the mito-
chondrial COI gene and a 47 1 bp fragment of the NDl gene, determined for
all four currently described species of Echinococcus, seven strains of E .
granulosus and two strains of E . multilocularis (Table 1). Published
sequence data are not available for the putative cervid and lion strains of
E . granulosus.
The data were analysed by maximum parsimony, using the branch and
bound algorithm of PAUP 3.1.1 (Swofford, 1993). All nucleotide positions
were considered and base changes weighted equally for analysis. Fasciola
hepatica and Ascaris s u m (sequence data obtained from GenBank and
aligned with the Echinocmcus sequences with the aid of the program
MacVectorTM(International Biotechnologies Inc.)) were used as outgroups
to provide the rooted trees shown in Figure 1.
Separate analysis of the COI and ND1 data sets produced quite different
minimum-length trees (cf Figure l a and lb), with little consensus in
higher-level structure (Figure lc). Analysis of the combined datasets
produced two minimum-length trees and a consensus of these two (Figure
Id) showed an almost identical topology to the ND1 tree. There has
recently been much dispute about the relative merits of consensus and
Combined approaches to obtaining an overall estimate of phylogeny from
two or more data sets (Miyamoto, 1985; Barrett et al., 1991; Bull et al.,
1993; de Queiroz, 1993). We have assumed that the COI and NDl
sequences, both being from the non-recombining mitochondrial genome,
cannot be regarded as providing independent characters and that a com-
bined approach should provide a more accurate estimate of phylogeny than
separate analysis of either sequence (de Queiroz, 1993).
We believe, therefore, that the tree shown in Figure Id is the best
estimate of phylogenetic relationships between OTUs of Echinococcus
available from currently published data. However, it should be regarded
as no more than a testable hypothesis of the true phylogeny. For many of
the OTUs, only a small number of isolates from a limited geographic range
have been sampled. More importantly, the data sets which have been
analysed are incomplete in that they do not include OTUs of uncertain
status, such as the cervid and lion strains of E. granulosus; inclusion of
these OTUs may alter the phylogeny. Fasciola hepatica and Ascaris suum
were used as outgroups in the analysis because they were the most closely
related helminths for which published DNA sequence data were available.
Effective character polarization requires more closely related outgroups,
preferably in the same family as Echinococcus (Maddison et al., 1984).
Finally, the phylogenetic analysis was based on sequences from only two
genes, both in the mitochondria1 genome. To increase the probability that
such gene trees accurately reflect the true evolutionary pathway of the
OTUs involved, requires sequences from a number of genes that have
evolved independently (Pamilo and Nei, 1988). Additional sequence data
are therefore needed from the nuclear genome of Echinococcus OTUs,
preferably from one of the more rapidly evolving regions of ribosomal
DNA (Hillis and Dixon, 1991). Bowles and McManus (1993a) state that
they are sequencing the internal transcribed spacer 1 region of the ribo-
somal DNA repeat unit, and this may provide data for a more definitive
phylogenetic analysis.
The uncertainty of our current understanding of phylogenetic relation-
GSH GSH
GTA GBU
GBU GTA
GHO GHO
VOG Gcr
MNO GCM
MEU GPI
OLI MNO
Gcr MEU
GCM VOG
GPI OLI
GSH GSH
GBU GBU
(ETA GTA
GHO GHO
Gcr Gcr
GCM GCM
GPI GPI
MNO MNO
MEU MEU
VOG VOG
OLI OLI
Figure 1 Phylogenetic trees identified using the branch and bound algorithm of
PAUP 3.1.1 (Swofford, 1993) on sequence data from regions of the mitochondria1
COI and ND1 genes (Bowles et al., 1992a; Bowles and McManus, 1993~).
Fasciola hepatica and Ascaris suum were used as outgroups for all trees. Numbers
at nodes represent percentage occurrence of clades in lo00 bootstrap replications
of the data. (a) Single minimum-length tree from analysis of 366 bp fragment of the
COI gene. Length of tree = 316 steps, consistency index = 0.84. (b) Single
minimum-length tree from analysis of 471 bp fragment of the ND1 gene. Length
of tree = 527 steps, consistency index = 0.784. (c) Strict consensus (Sokal and
Rohlf, 1981) between the trees in (a) and (b). (d) Strict consensus of two minimum-
length trees from analysis of combined COI and NDl data. Length of trees = 853
steps, consistency index = 0.80. OTU codes as in Table 1.
ships within the genus Echinococcus is reflected in the poor support from
bootstrap analyses (Felsenstein, 1985) for much of the higher-level struc-
ture shown in Figure Id. Clearly, clades that appear in only 18%, 19% or
even 48%-54% of bootstrap samples should be viewed with suspicion.
Nevertheless, we believe that there is enough well-supported structure at
lower levels in the tree to warrant the ranking of some clades as evolu-
tionary species. The groupings shown in Figure Id which were consistently
supported by parsimony analysis of bootstrap samples were also robust to
phylogenetic distance analyses (results not shown).
5. DELIMITATION OF EVOLUTIONARV SPECIES
It seems likely that current classification within the genus Echinococcus is

not compatible with historical relationships between taxa. Phylogenetic
analysis of DNA sequence data provides no support for the concept that
E. grunulosus is a monophyletic group and it therefore cannot be con-
sidered an evolutionary species. We believe that, on genetic, morpho-
logical and ecological grounds, there is strong evidence for the existence
of seven separate species within the genus. In the following sections, we
enumerate these putative species and provide some historical background
to their nomenclature (Table 2). The manner in which we proceed is to
begin with the most nested OTUs from the tree shown in Figure Id and
work outwards. At each step, we determine whether sister clades possess
genetic or demographic exchangeability; if they do not, then they are
ranked as separate species. Given the uncertainty of higher-level structure
in the tree shown in Figure Id, we also comment, where appropriate, on
the potential exchangeability with OTUs in other clades. We do not
propose to review the taxonomy of Echinococcus since this has been
done comprehensively on previous occasions (Verster, 1965; Rausch,
1967; Kumaratilake and Thompson, 1982; Thompson and Lymbery,
1988), and detailed information on the correct nomenclature for species
and subspecies is well documented. Fortunately, as emphasized previously
(Kumaratilake and Thompson, 1982; Thompson and Lymbery, 1988), the
majority of intraspecific variants of Echinococcus have been recognized as
species or subspecies in the past and, in most cases, accurate and detailed
descriptions are available.
5.1. Species 1 (Echinococcussp.)
The pig and camel strains of E. grunulosus (GPI and GCM in Figure 1)
were invariably monophyletic in parsimony analyses of mtDNA sequence
Table 2 Putative evolutionary species in the genus Echinococcus.
~~ ~
Suggested Known Known

Evolutionary taxonomic definitive intermediate Probablea geographic
species designation hosts hosts distribution Synonyms Strains
~~ ~~
Species 1 E. sp.? Dog Pigs, humans? Europe, Russia, South E. granulosus pig strain Pig strain?
America
Species 2 E. ortleppi Dog Cattle, buffalo, Europe, Africa, India, E. granulosus ortleppi,
humans Sri Lanka. Russia E. granulosus cattle
Strain
Species 3 E. equinus Dog Horses and other Europe, Middle East, E. granulosus equinus,
equines South Africa (New E. granulosus horse
Zealand? USA?) strain
Species 4 E. multilocularis Fox, dog, Rodents, pigs, Europe, North America, E. sibiricensis European strain,
cat horses, humans Canada, Japan, China North American
strain, Alaskan
strain? Hokkaido
Strain?
Species 5 E. vogeli Bush dog Rodents, humans South America
Species 6 E. oligarthrus Felines Rodents, humans South America E. pampeanus, E. cruzi
Species 7 E. granulosus Dog, fox, Sheep, cattle, Australia, Europe, E. patagonicus, E . Common sheep
dingo, pigs, goats, USA, New Zealand, cepanazoi, E. g. strain, Tasmanian
jackal, buffalo, camels, Africa, China, Middle granulosus, E. g. sheep strain,
hyena macropods, East, Asia, South newzealandensis Buffalo strain?
humans America, Russia
a The geographic range of some species still needs to be fully defined.

data. There is no evidence that these OTUs occur in sympatry and their
potential for exchangeability must be inferred from genetic, morphological
and ecological data. Morphological studies have suggested affinities
between the taxa (Eckert et al., 1993), but genetic data are equivocal.
Sequence analysis of mitochondrial COI and ND1 genes suggests very
close genetic similarity (0.003, 0.006 base substitutions per nucleotide,
respectively, as estimated by the method of Jukes and Cantor (1969)
from the data of Bowles et al. (1992a) and Bowles and McManus
(1993c)), as do some RFLP analyses of rDNA (McManus and Rishi,
1989; Bowles and McManus, 1993b). Eckert et al. (1993), however, found
much greater genetic differences from RFLP analyses of rDNA and an
uncharacterized fragment of genomic DNA (0.06, 0.01 base substitutions
per nucleotide respectively, as estimated by the method of Nei and Li
(1979)). The camel isolates used in these studies were from different
geographic areas, however, and, as discussed previously (Section 3.1.6),
epidemiological evidence suggests the possible occurrence of different
OTUs in cycles involving camels. We believe further data are required
before a decision can be reached on the taxonomic status of the form of
Echinococcus found in camels.
At this point, we consider the OTU currently known as the pig strain of
E. granulosus to be a valid evolutionary species (Table 2). As far as we are
aware, this taxon has not been previously named, although the descriptions
given by Vogel (1957) and Verster (1965) for E. granulosus of pig origin
could be used as the type.
5.2. Species 2 (Echinococcusortleppi?)
The cattle strain of E. granulosus (GCT in Figure 1) forms a well-

supported monophyletic group with Species 1. They also occur sympatri-
cally (e.g. Vogel, 1957; Hahn et al., 1988; Worbes et al., 1989) and
maintain morphological, developmental (Thompson et al., 1984; Eckert
et al., 1993) and genetic differences ranging from 0.051 to 0.078 base
substitutions per nucleotide for mitochondrial genes (calculated from the
data of Bowles et al. (1992a) and Bowles and McManus (1993~))and 0.066
to 0.104 base substitutions per nucleotide for genomic sequences (Eckert et
al., 1993).
The OTUs, therefore, must be regarded as separate evolutionary species
(Table 2). According to morphological and genetic analyses, the cattle
strain occurs throughout Europe as well as parts of Africa and Asia
(Section 3.1.3). Thus, the most appropriate species name would appear
to be E. ortleppi following Lopez-Neyra and Soler Planas’ (1943) designa-
tion for adult worms originally described by Ortlepp (1934) from the type
VARIATIONIN ECH~NOCOCCUSTOWARDS A TAXONOMIC REVISION OF THE GENUS 165
locality in South Africa, and which Verster (1965), Kumaratilake (1982)

and Thompson ef al. (1984) considered to be of cattle origin.
5.3. Species 3 (Echinococcus equinus?)
The horse strain of E. granulosus (GHO in Figure 1) does not consistently

group with any other OTUs in parsimony analyses of DNA sequence data.
It maintains genetic, morphological and ecological differences in sympatry
with its presumed closest relatives, Species 1 and Species 2, and also with
the sheep strain of E. granulosus and with E . multilocularis (Kumaratilake
er al., 1986; McManus and Rishi, 1989; Bowles et al., 1992a; Bowles and
McManus, 1993b, c; Eckert et al., 1993), and should therefore be regarded
as a separate evolutionary species (Table 2), thus confirming previous
speculation (Rausch in Schantz, 1982; Thompson and Lymbery, 1988,
1991; Lymbery, 1992; Bowles and McManus, 1993a, b). This taxon was
described from UK material as a subspecies, E. g. equinus, by Williams and
Sweatman (1963), but subspecific status was invalidated by Rausch (1967).
Under the International Code of Zoological Nomenclature, equinus would
appear to have priority as a species-group name, and Williams and
Sweatman (1963) provided a detailed aad accurate description of the
parasite of horse/dog origin from the type locality in Britain.
5.4. Species 4 (Echinococcus multilocularis)
The two strains of E. multilocularis (MEU and MNO in Figure 1) fonn a

well-supported monophyletic group. They maintain genetic, morphological
and ecological differences in sympatry with Species 1, 2 and 3, and with
the sheep strain of E. granulosus (Eckert and Thompson, 1988; Thompson
and Lymbery, 1988, 1991; Bowles et al., 1992a; Bowles and McManus,
1993b, c; Eckert et al., 1993) and are clearly a separate evolutionary
species.
There is evidence of morphological and biological differences between
populations of E. multilocularis from Eurasia and North America (Eckert
and Thompson, 1988; Bartel et al., 1992), but genetic differences found to
date are very small (0.004-0.006 base substitutions per nucleotide in
mtDNA genes, estimated from the data of Bowles et al. (1992a) and
Bowles and McManus (1993~))and do not correspond to geographic
divisions. We consider that the different populations of E. multilocularis
are capable of genetic or demographic exchangeability and therefore con-
stitute a single evolutionary species (Table 2). On the basis of their small,
but consistent genetic differences and differences in epidemiologically
significant traits such as prepatent period (Eckert and Thompson, 1988),

populations of E. multilocularis in Europe and North America should be
regarded as different strains by the definition of Thompson and Lymbery
(1988). Further studies are required on Chinese and Japanese populations.
5.5. Species 5 and 6 (Echinococcus vogeli and Echinococcus

oligarthrus)
These OTUs (VOG and OLI in Figure 1) form a reasonably well supported
monophyletic group. They maintain major genetic, morphological and
.ecological differences in sympatry and therefore must be regarded as
separate evolutionary species (Table 2). Neither species consistently
groups with other OTUs in phylogenetic analyses and they are both quite
distinct genetically from all other species (for E. ,vogeli 0.060.184, for
E. oligarthrus 0.093-0.184 base substitutions per nucleotide in mitochon-
drial genes, calculated from the data of Bowles et al. (1992a) and Bowles
and McManus (1993~)).
5.6. Species 7 (Echinococcusgranulosus)
The sheep, Tasmanian sheep and buffalo strains of E. granulosus (GSH,

GTA and GBU in Figure l), are monophyletic and genetically distinct from
all other species (0.08 1-0.184 base substitutions per nucleotide in mito-
chondrial genes, calculated from the data of Bowles et al. (1992a) and
Bowles and McManus (1993~)).Genetic differences between the three
OTUs are minor. The data of Bowles et al. (1992a) and Bowles and
McManus (1993~)indicate 0 . 0 0 8 base substitutions per nucleotide
separating them in mitochondria1 genes. The three OTUs could not be dis-
tinguished by RFLP analysis of rDNA (McManus and Rishi, 1989; Bowles
and McManus, 1993b). Allozyme studies by Lymbery and Thompson
(1988) suggest 0.012 base substitutions per enzyme locus (estimated by
the method of Nei (1978)) separating Australian mainland and Tasmanian
populations of Echinococcus in sheep, well within the range expected for
conspecific populations (Thorpe, 1982, 1983).
These data suggest that the OTUs are capable of genetic or demographic
exchangeability and we regard them as a single evolutionary species (Table
2). Although the species name derives from Batsch’s (1786) early descrip-
tions of hydatid cysts in sheep, the classical description of E. granulosus
was given by Vogel(l957) from a type locality in Germany. Unfortunately,
however, Vogel’s description was based on adult worms of German pig/
dog origin. Such a description can not be considered to be representative of
E. grunulosus, since the morphological characteristics of Vogel’s material

closely corresponds to the pig strain (Kumaratilake and Thompson, 1982;
Eckert ef ul., 1993) which occurs in Europe (Section 3.1.5) and is almost
certainly a distinct species. Consequently, the description given by
Williams and Sweatman (1963) for the subspecies E. g. grunulosus, which
is b a s 4 on material of New Zealand sheep/dog origin, is the most
appropriate for this species.
Despite the close genetic similarity between the sheep and Tasmanian
sheep OTUs, they differ morphologically and in prepatent period and cyst
developmentrate (Kumaratilake and Thompson, 1983,1984a; Kumaratilake
ef g l . , 1983) and should be regarded as separate strains (Table 2). As
indicated in Section 3.1.7, the status of forms affecting buffalo requires
further study.
6. CONCLUSIONS
We believe that an evolutionarily sound species-level classification in the

genus Echinococcus, consistent with historical relationships between popu-
lations and the action of population genetic processes, is essential for the
effective control of hydatid disease. Without the stability of nomenclature
and the predictive powers provided by such a classification, we can be
seriously misled in our attempts to apply basic research data to the
practicalities of disease control. For example, much effort has been
expended in developing immunological and molecular techniques for the
diagnosis of hydatid disease in people and other intermediate hosts, or of
adult worms in definitive hosts (Rickard and Lightowlers, 1986; McManus,
1990; Pawlowski, 1992; Lightowlers and Gottstein, 1994). For most of
these techniques, however, there is evidence of a lack of species specificity
and of poor diagnostic sensitivity (Leggatt et u1.,1992; Gasser ef ul., 1993;
Lightowlers and Gottstein, 1994). These problems undoubtedly reflect the
current, biologically misleading taxonomy in the genus Echinococcus,
especially the diversity of evolutionary species grouped under the binomen
Echinococcus grunulosus.
As was predicted in the earlier review (Thompson and Lymbery, 1988),
the discriminatory power of biochemical and molecular characterization
techniques has given us the data to propose a taxonomic revision of the
genus Echinococcus. The way is now open for detailed comparative studies
to be undertaken in a number of endemic areas in order to determine the
geographic distribution and uniformity of the species we have nominated
so that formal taxonomic designations can be confirmed. In addition,
there is an urgent need for the molecular characterization of strains of
E. multilocularis as well as strains of Echinococcus in lions and cervids, so

that this taxonomic revision can be completed.
In the near future, melecular techniques are also likely to make
significant inroads into elucidating the functional nature of variation in
Echinococcus, and should provide clues as to how strains adapt to
different hosts and the regulatory mechanisms that govern genetic and
biochemical interactions between host and parasite (Thompson, 1994).
ACKNOWLEDGEMENTS
Our research is supported by the Australian Research Grants Scheme and

World Health Organization. Thanks to Sue Lyons for her efficient and
accurate typing.
REFERENCES
Abraham, J., Pillai, K.M. and Iyer, R.P. (1980). Fertility rate in hydatid cysts in
domestic animals. Kerala Journal of Veterinary Science 11, 155-158.
Ballek, D. (199 1). Zum Vorkommen von Echinococcus multilocularis und anderen
Zestoden und Nematoden beim Rotfuchs (Vulpes vulpes L.) in den Regierungs-
bezirken Amsberg, Detmold and Kassel. Inaugural Dissertation, Tierarztliche
Hochschule Hannover, Hannover, Germany.
Barrett, M., Donoghue, M.J. and Sober, E. (1991). Against consensus. Systematic
Zoology 40, 486-493.
Bartel, M.H., Seesee, F.M. and Worley, D.E. (1992). Comparison of Montana and
Alaska isolates of Echinococcus multilocularis in Gerbils with observations on
the cyst growth, hook characteristics and host response. Journal of Parasitology
78, 528-529.
Batsch, A.J.G.C. (1786). Naturgeschichte der Bandwurmgattung Uberhaupt und
i b e r Arten insbesondere, nach den neuen Beobachtungen in einem systema-
tischen Auszuge. Halle.
Bowles, J. and McManus, D.P. (1991). Molecular characterisation of Echinococcus.
Archivos De La Hidatidosis 30, 55-63.
Bowles, J. and McManus, D.P. (1993a). Molecular variation in Echinococcus. Acta
Tropica 53, 291-305.
Bowles, J. and McManus, D.P. (1993b). Rapid discrimination of Echinococcus
species and strains using a polymerase chain reaction-based RFLP method.
Molecular and Biochemical Parasitology 57, 23 1-240.
Bowles, J. and McManus, D.P. (1993~).NADH dehydrogenase 1 gene sequences
compared for species and strains of the genus Echinococcus. International
Journal for Parasitology 23,969-972.
Bowles, J., Blair, D. and McManus, D.P. (1992a). Genetic variants within the
genus Echinococcus identified by mitochondria1 DNA sequencing. Molecular

and Biochemical Parasitology 54, 165-174.
Bowles, J., van Knapen, F. and McManus, D.P. (1992b). Cattle strain of Echino-
coccus granulosus and human infection. Lancet 339, 1358.
Brochier, B., Coppens, P., Losson, B., Aubert, M.F.A., Bauduin, B., Barrat, M.J.,
Costy, F., Peharpre, D., Pouplard, L. and Pastoret, P.-P. (1992). Enquete sur
l’infestation du renard roux (Vulpes vulpes),par Echinococcus multilocularis en
province de Luxembourg (Belgique). Annales de Medecine Veterinaire 136,
497-501.
Bull, J.J., Huelsenbeck, J.P., Cunningham, C.W., Swofford, D.L. and Waddell, P.J.
(1993). Partitioning and combining data in phylogenetic analysis. Systematic
Biology 42, 384-397.
Canieron, T.W.M. (1960). The incidence and diagnosis of hydatid cyst in Canada.
Echinococcus granulosus var. canadensis. Parassitologia 2, 38 1-390.
Clarkson, M.J. and Walters, T.M.H. (1991). The growth and development of
Echinococcus granulosus of sheep origin in dogs and foxes in Britain. Annals
of Tropical Medicine and Parasitology 85, 53-61.
Constantine, C., Lymbery, A.J., Thompson, R.C.A. and Obendorf, D.L. (1991).
The origin of a new focus of infection with Echinococcus granulosus in
Tasmania. International Journal for Parasitology 21, 959-962.
Constantine, C.C., Thompson, R.C.A., Jenkins, D.J., Hobbs, R.P. and Lymbery,
A.J. (1993). Morphological characterization of adult Echinococcus granulosus
as a means of determining transmission patterns. Journal of Parasitology 79,
57-61.
Cruz-Reyes, A., Tavizon, P. and D’Alessandro, A.D. (1990). Development of adult
Echinococcus granulosus from hydatids of naturally infected pigs. Bulletin De
La Societe Francaise de Parasitologie 8, Suppl. No. 1, 237.
de Queiroz, A. (1993). For consensus (sometimes). Systematic Biology 72,368-372.
De Rycke, P.H. (1968). Het voorkomen van steriele hydatiden bij runderen en de
subspeciate van Echinococcus granulosus. Vlaams Diergeneeskundig Tijdschrift
37, 101-108.
Dissanaike, A.S. (1957). Some preliminary observations on Echinococcus infection
in local cattle and dogs. Ceylon Medical Journal 4, 69-75.
Donoghue, M.J. (1985). A critique of the biological species concept and
recommendations for a phylogenetic alternative. Bryologist 88, 172-1 8 1.
Eckert, J. and Thompson, R.C.A. (1988). Echinococcus strains in Europe: a review.
Tropical Medicine and Parasitology 39, 1-8.
Eckert, J., Thompson, R.C.A., Michael, S.A., Kumaratilake, L.M. and El-Sawah,
H.M. (1989). Echinococcus granulosus of camel origin: Development in dogs
and parasite morphology. Parasitology Research 75, 536-544.
Eckert, J., Thompson, R.C.A., Lymbery, A.J., Pawlowski, Z.S., Gottstein, B. and
Morgan, U.M. (1993). Further evidence for the occurrence of a distinct strain of
Echinococcus granulosus in European pigs. Parasitology Research 79, 42-48.
Felsenstein, J. (1985). Confidence limits on phylogenies: an approach using the
boostrap. Evolution 39, 783-79 1.
French, C.M., Nelson, G.S. and Wood, M. (1982). Hydatid disease in the Turkana
district of Kenya. I. The background to the problem with hypotheses to account
for the remarkably high prevalence of the disease in man. Annals of Tropical
Medicine and Parasitology 76, 425-437.
Gasser, R.B., Jenkins, D.J., Paolillo, E., Parada, L., Cabsera, P. and Craig, P.S.
170 A.C.A. THOMPSON ET AL
( 1993). Serum antibodies in canine echinococcosis. International Journal for

Gebreel, A.O., Gilles, H.M. and Prescott, J.E. (1983). Studies on the sero-
epidemiology of endemic diseases in Libya. Annals of Tropical Medicine
and Parasitology 77, 391-397.
Gill, H.S. and Venkateswara, R. (1967). Incidence and fertility rate of hydatid
cysts in Indian buffalo (Bos bubalus). Bulletin de I' OfSlce Internationale des
Epizooties 69, 989-997.
Gottstein, B. (199 1). Echinococcus multilocularis: antigenic variance between
different parasite isolates. Parasitology Research 77, 359-36 1.
Gottstein, B., Eckert, J. and Fey, H. (1983). Serological differentiation between
Echinococcus granulosus and E. multilocularis infections in man. Zeitschrifr fur
Parasitenkunde 69, 347-356.
Graber, M. and Thal, J. (1980). L'echinococcose des artiodactyles sauvages de la
' Republique Centrafricaine: existence probable d'un cycle lion-phacochere.
Revue Elevage Medecine Veterinaire Pays Tropicaux 33, 5 1-59.
Hahn, E.,Horchner, F., Huwer, M. and Sanft, S. (1988). Zur Frage der Stamme-
spezifitat von Echinococcus granulosus (Rind). in Deutschland. Wiener Tier-
arztliche Monatsschrift 75, 476-479.
Harrison, L.J.S., Le Riche, P.D. and Sewell, M.M.H. (1986). Variations in Echino-
coccus granulosus of bovine origin identified by enzyme electrophoresis. Tro-
pical Animal Health and Production 18, 48-50.
Hijjawi, N.S., Abdel-Hafez, S.K. and Al-Yaman, F.M. (1992). In vitro culture of
the strobilar stage of Echinococcus granulosus of sheep and donkey origin from
Jordan. Parasitology Research 78, 607-6 16.
Hildreth, M.B., Johnson, M.D. and Kazacos, K.R. (1991). Echinococcus multi-
locularis: a zoonoses of increasing concern in the United States. Compendium on
Continuing Education for the Practicing Veterinarian 13, 727-74 1.
Hillis, D.M. and Bull, J.J. (1993). An empirical test of bootstrapping as a
method for assessing confidence in phylogenetic analysis. Systematic Biology
42, 182-192.
Hillis, D.M.and Dixon, M.T. (1991). Ribosomal DNA: molecular evolution and
phylogenetic inference. Quarterly Review of Biology 66, 41 1 4 5 3 .
Hobbs, R.P., Lymbery, A.J. and Thompson, R.C.A. (1990). Rostellar hook
.morphology of Echinococcus granulosus (Batsch, 1786) from natural and
experimental Australian hosts and its implications for strain recognition. Para-
sitology 101, 273-28 1.
Hope, M., Bowles, J. and McManus, D.P. (1991). A reconsideration of the
Echinococcus granulosus strain situation in Australia following RFLP analysis
of cystic material. International Journal for Parasitology 21, 47 1-475.
Hu, W., Yan-ling, C., Hai-yu, Y.and Hai-yan, W. (1993). Morphological observa-
tion on the adults of Echinococcus granulosus from the experimental infected
dogs used different origin protoscoleces in Qinghai plateau. Endemic Diseases
Bulletin 8, 6-10.
Huldt, G., Johansson, G.S.O. and Lantto, S. (1973). Echinococcosis in northern
Scandinavia. Archives of Environmental Health 26, 35-40.
Irshadullah, M., Nizami, W.A. and Macpherson, C.N.L. (1989). Observations on
the suitability and importance of domestic intermediate hosts of Echinococcus
granulosus in Uttar Pradesh, India. Journal of Helminthology 63, 3 9 4 5 .
Islam, A.W.M.S. (1982). The prevalence of hydatid disease in buffaloes in
Bangladesh. Annals of Tropical Medicine and Parasitology 76, 623-626.
Janssen, D., De Wit, M. and De Rycke, P.H. (1990). Hydatidosis in Belgium:

analysis of larval Echinococcus granulosus by SDS-PAGE and Western blotting.
Annales de la Societe Belge de Medecine Tropicale 70, 121-129.
Jorgensen, R.J. and Hansen, J.W. (1984). Echinococcosis: a rare infection in
Danish livestock. In “Some Important Parasitic Infections in Bovines Con-
sidered from Economic and Social (Zoonosis) Points of View” (J. Euzeby and
J. Gevrey, eds), pp. 115-120. Commission of the European Communities,
Luxemburg.
Jukes, T.H. and Cantor, C.R. (1969). Evolution of protein molecules. In
“Mammalian Protein Metabolism” (H.N. Munro, ed.), pp. 21-1 32. Academic
Press, New York.
Kammerer, W.S. and Schantz, P.M. (1984). Long term follow-up of human hydatid
disease (Echinococcus granulosus) treated with a high-dose mebendazole
regimen. American Journal of Tropical Medicine and Hygiene 33, 132-1 37.
Kamiya, M. (1988). Infectious diseases transmitted by dogs to humans. Asian
Medical Journal 31,87-93.
Kawase, S. and Yagi, K. (1985). Studies on experimental multilocular hydatidosis
3. Characterization of Echinococcus multilocularis isolated in Abashiri region,
north-eastem Hokkaido, Japan. Japanese Journal of Parasitology 34, 8 1 (In
Japanese).
Kumaratilake, L.M. (1982). Aspects of the speciation of Echinococcus granulosus
in Australia. PhD Thesis, Murdoch University.
Kumaratilake, L.M. and Thompson, R.C.A. (1982). A review of the taxonomy
and speciation of the genus Echinococcus Rudolphi 1801. Zeitschrifi fur
Kumaratilake, L.M. and Thompson, R.C.A. (1983). A comparison of Echinococcus
granulosus from different geographical areas of Australia using secondary cyst
development in mice. International Journal for Parasitology 13, 509-5 15.
Kumaratilake, L.M. and Thompson, R.C.A. (1984a). Morphological characterisa-
tion of Australian strains of Echinococcus granulosus. International Journal for
Parasitology 14,467477.
Kumaratilake, L.M. and Thompson, R.C.A. (1984b). Biochemical characterisation
of Australian strains of Echinococcus granulosus by isoelectric focusing of
soluble proteins. International Journal for Parasitology 14,58 1-586.
Kumaratilake, L.M., Thompson, R.C.A. and Dunsmore, J.D. (1 979). Intraspecific
variation in Echinococcus: a biochemical approach. Zeitschrift fur Parasiten-
kunde 60, 291-294.
Kumaratilake, L.M., Thompson, R.C.A. and Dunsmore, J.D. (1983). Comparative
strobilar development of Echinococcus granulosus of sheep origin from different
geographical areas of Australia in vivo and in vitro. International Journal for
Kumaratilake, L.M., Thompson, R.C.A. and Eckert, J. (1986). Echinococcus
granulosus of equine origin from different countries possess uniform morpho-
logical characteristics. International Journal for Parasitology, 16,529-540.
Leggatt, G.R., Yang, W. and McManus, D.P. (1992). Serological evaluation of
the 12 kDa subunit of antigen B in Echinococcus granulosus cyst fluid by
immunoblot analysis. Transactions of the Royal Society for Tropical Medicine
and Hygiene 86, 189-192.
Liance, M., Bresson-Hadni, S., Vuitton, D., Bretagne, S. and Houin, R. (1990).
Comparison of the viability and developmental characteristics of Echinococcus
multilocularis isolates from human patients in France. International Journal for

Lightowlers, M.W. and Gottstein, B. (1994). Echinococcosisihydatidosisantigens,
immunological and molecular diagnosis. In “Echinococcus and Hydatid
Disease” (R.C.A. Thompson and A.J. Lymbery, eds), CAB International,
Wallingford, UK (in press).
Lightowlers, M.W., Rickard, M.D., Honey, R.D., Obendorf, D.L. and Mitchell, G.F.
(1984). Serological diagnosis of Echinococcus granulosus infection in sheep
using cyst fluid processed by antibody affinity chromatography. Australian
Lopez-Neyra, C.R. and Soler Planas, M.A. (1943). Revision del genero Echino-
coccus Rud. y descripcion de una especie nueva parasita intestinal del perro en
Almeria. Revista Iberica Parasitologia 3, 169-1 94.
.
Lymbery, A.J. (1992). Interbreeding, monophyly and the genetic yardstick: species
concepts in parasites. Parasitology Today 8, 208-2 11.
Lymbery, A.J. (1994). Genetic diversity, genetic differentiation and speciation in
the genus Echinococcus Rudolphi 1801. In “&hinococcus and Hydatid
Disease” (R.C.A. Thompson and A.J. Lymbery, eds), CAB International,
Wallingford, UK (in press).
Lymbery, A.J. and Thompson, R.C.A. (1988). Electrophoretic analysis of genetic
variation in Echinococcus granulosus from domestic hosts in Australia.
International Journal for Parasitology 18, 803-8 11.
Lymbery, A.J., Thompson, R.C.A. and Hobbs, R.P. (1990). Genetic diversity and
genetic differentiation in Echinococcus granulosus (Batsch, 1786) from
domestic and sylvatic hosts on the mainland of Australia. Parasitology 101,
283-289.
Macchioni, G., Arispici, M., Lanfranchi, P. and Testi, F. (1987). Experimental
infection of sheep and monkeys with the camel strain of Echinococcus
granulosus. In “Helminth Zoonoses” ( S . Geerts, V. Kumar and J. Brandt, eds),
pp. 24-28. Martinus Nijhoff, Dordrecht.
Macpherson, C.N.L. (1986). Echinococcus infections in wild animals in Africa. In
“Wildlife/Livestock Interfaces on Rangelands” (S. MacMillan, ed). pp. 73-78.
Inter-African Bureau for Animal Resources, Nairobi, Kenya.
Waddison, W.P., Donoghue, M.J. and Maddison, D.R. (1984). Outgroup analysis
and parsimony. Systematic Zoology 33, 83-103.
Mayr, E. (1942). “Systematics and the Origin of Species”. Columbia University
Press, New York.
McKitrick, M.C. and Zink, R.M. (1988). Species concepts in ornithology. Condor
90, 1-14.
McManus, D.P. (1990). Characterisation of taeniid cestodes by DNA analysis.
Revue. Scientifrque et technique Ofice Internationale des Epizootics 9,489-5 10.
McManus, D.P. and Bryant, C. (1994). Biochemistry, physiology and molecular
biology of Echinococcus. In “Echinococcus and Hydatid Disease” (R.C.A.
Thompson and A.J. Lymbery, eds). CAB International, Wallingford (in press).
McManus, D.P. and Rishi, A.K. (1989). Genetic heterogeneity within
Echinococcus granulosus isolates from different hosts and geographical areas
characterised with DNA probes. Parasitology 99, 17-29.
McManus, D.P. and Smyth, J.D. (1982). Intermediary carbohydrate metabolism in
protoscoleces of Echinococcus granulosus (horse and sheep strains) and E.
multilocularis. Parasitology 84, 35 1-366.
McManus, D.P., Thompson, R.C.A. and Lymbery, A.J. (1989). Comment on the
status of Echinococcus granulosus in the UK. Parasitology Today 5, 365-368.
Mishler, B.D. and Donoghue, M.J. (1982). Species concepts: a case for pluralism.
Systematic Zoology 31, 49 1-503.
Miyamoto, M.M. ( 1 985). Consensus cladograms and general classifications.
Cladistics 1, 1 86- 1 89.
Morgan, U.M., Constantine, C.C., Lymbery, A.J. and Thompson, R.C.A.
Characterisation of Echinococcus using RAPD analysis. Submitted for
publication.
Nadler, S.A. ( 1987). Genetic variability in endoparasitic helminths. Parasitology
Today 3, 154-155.
Nei, M. (1978). Estimation of average heterozygosity and genetic distance from a
small number of individuals. Genetics 89, 583-590.
Nei, M. and Li, W.H. (1979). Mathematical model for studying genetic variation in
terms of restriction endonucleases. Proceedings of the National Academy of
Sciences of the USA 76, 5269-5273.
Nozais, J.P. (1987). Hypotheses sur le role du Sahara prehistorique dans la
repartition de certaines affections parasitaires et hematologuiques. Bulletin de
la Societe de Pathologie Emtiyue et de ses Filiales 80, I2 1-1 3 I .
O’Hara, R.J. ( 1993). Sytematic generalization, historical fate, and the species
problem. Systematic Biology 42, 23 1-246.
Okamoto, M., Fujita, O., Arikawa, J., Kurosawa, T., Oku, Y. and Kamiya, M.
( 1992). Natural Echinocoiru.s multilocirlaris infection in a Norway rat, Rattus
norvegicirs, in southern Hokkaido, Japan. International Journal f o r Parasitology
22, 681-684.
Ortlepp, R.J. ( 1934). Echinococciu in dogs from Pretoria and vicinity. Onder-
stepoort Joional of Veterinary Science 3. 97- 108.
Ortlepp. R.J. ( 1937). South African helminths. Part I. Otirlrrstepoort Journal of
Veterinary Science 9, 3 I 1-366.
Pamilo, P. and Nei. M. (1988). Relationships between gene trees and species trees.
Molecular- Biology anti Eiwlutiori 5 . 568-583.
Pandey, V.S. (1972). Observations on the morphology and biology of Echino-
i ~ o ( ~ c i granir1osu.s
rs (Batsch. 1786) of goat-dog origin. .loirrnul of Helminthology
46, 2 19-233.
Pawlowski, Z.S. ( 1992). Ei.hitioc.oc.c.ir.s and cysticercosis. Current Opinion in
1nfc~ctioir.sDiseci.sc,s 5. 3 19-325.
Ptister. T. and Frank, W. (1988). Experimentelle Untersuchungen zur Empfang-
lichkeit des Schweines fur E t ~ h i t i o c ~ o c ~ cniultiloc~irlar-is.
~us Mitteilirrigen der
Ostcr~rcic~hisc~heti Ge.seIlschuft,fiir Tropenniedi~iriirnd Porasitologie 10, 1 03- 1 1 4.
Rausch. R.L. ( 1967). A consideration of infraspecific categories in the genus
Ec~lritioc~oc~c~u.s Rudolphi, I80 I (Cestoda: Taeniidae). Joiirnal qf Parasitology
53. 4 8 4 4 9 I.
Rausch. R.L. ( 1986). Life-cycle patterns and geographic distribution of Echino-
c ~ ~ i xspecies.s In ”The Biology of E~~/iirioc~oc~i~rrs and Hydatid Disease”.
(R.C.A. Thompson. ed.), pp. 44-80. George Allen and Unwin. London.
Rausch. R.L. ( 1994). Life cycle patterns and geographic distribution of Ediino-
i ~ o ( ~ species.
xs I n “E(,hitioc.oc,c,irsand Hydatid Disease”. (R.C.A. Thompson and
A.J. Lymbery. eds). CAB International. Wallingford, U K ( i n press).
Rickard. M.D. and Lightowlers, M.W. ( 1986). lmmunodiagnosis of hydatid
disease. In “The Biology of Ei,liirioc,oc,c,ir.s and Hydatid Disease” (R.C.A.
Thompson, ed.). pp. 217-249. George Allen and Unwin. London.
Said, I.M., Abdel-Hafez, S.K. and Al-Yaman, F.M. (1988). Morphological varia-
tion of Echinococcus granulosus protoscoleces from hydatid cysts of human and
various domestic animals in Jordan. International Journal for Parasitology 18,
11 11-1 114.
Saimot, A.G., Meulemans, A., Hay, J.M., Mohler, J., Manuel, C. and Coulaud, J.P.
(1981). Etude pharmacocinetique du flubendazole au cours de l’hydatidose
humaine a E . granulosus., Resultats preliminaies. Nouvelle Presse Medicale
10, 3121-3124.
Sanyal, P.K. and Sinha, P.K. (1983). A note on the prevalence of hydatidosis in
cattle and buffalo in West Bengal. Haryana Veterinarian 22, 47-50.
Schantz, P.M. (1982). Echinococcosis. In “CRC Handbook Series in Zoonoses”
(J. Steele, ed.), Section C, 1, pp. 231-277. CRC Press, Boca Raton.
Schantz, P.M. (199 1). Parasitic zoonoses in perspective. International Journal for
Schantz, P.M., Van den Bossche, H. and Eckert, J. (1982). Chemotherapy for larval
echinococcosis in animals and humans: report of a workshop. Zeitschrgt fur
Shabovskaya, E.A., Bulgakov, V.A., Ponomareva, V.E., Dariko, O.P., Voloshchuk,
S.D. and Kikot, V.11. (1989). Hydatidosis in the Ukranian SSR. Meditsinskaya
Parazitologiya i Parazitarnye Bolezni 6, 49-5 1 (In Russian).
Siles-Lucas, M., Cuesta-Bandera, C. and Cesar-Benito, M. (1993). Random ampli-
fied polymorphic DNA technique for speciation studies of Echinococcus
granulosus. Parasitology Research 79, 343-345.
Singh, B.P., Srivastava, V.K. and Sharma Deorani, V.P. (1988). Pig hydatidosis in
Uttar Pradesh. Veterinary Record 123, 299-300.
Skvortsova, F.K. and Artemenko, Yu.G. (1987). Morphological characteristics of
Echinococcus granulosus strains in southern Ukraine. Byulleten’ Vsesoyuznogo
Instituta Gel’mintologii im K.I. Skryabina 47, 69-72 (In Russian).
Smyth, J.D. (1990). Parasitological serendipity: from Schistocephalus to Echino-
coccus. International Journal for Parasitology 20, 4 11-423.
Smyth, J.D. and Smyth, M.M. (1969). Self insemination in Echinococcus
granulosus in vivo. Journal of Helminthology 43, 383-388.
Sneath, P.H.A. and Sokal, R.R. (1973). “Numerical Taxonomy”. W.H. Freeman,
San Francisco.
- Sokal, R.R. and Rohlf, F.J. (198 1). Taxonomic congruence in the Leptopodomorpha
re-examined. Systematic Zoology 30, 309-325.
Storandt, S.T. and Kazacos, K.R. (1993). Echinococcus multilocularis identified in
Indiana, Ohio, and east-central Illinois. Journal of Parasitology 79, 301-305.
I Swofford, D.L. (1993). “PAUP: Phylogenetic Analysis Using Parsmony Version
3.1 ”. Illinois Natural History Survey, Champaign, Illinois.
Templeton, A.R. (1989). The meaning of species and speciation: a genetic
perspective. In “Speciation and its Consequences” (D. Otte and J.A. Endler,
eds), pp. 3-27. Sinauer, Sunderland, Massachusetts.
Thompson, R.C.A. (1988). Intraspecific variation and epidemiology. In:
“Parasitology in Focus”. (H. Mehlhorn, ed.), pp. 3 9 1 4 1 1, Springer, Berlin.
Thompson, R.C.A. (1991). Echinococcus and Giardia: variation on a theme.
International Journal for Parasitology 21, 29 1-297.
Thompson, R.C.A. (1992). Echinococcosis/Hydatidosisin Australia. In “Zoonoses.
Proceedings 194. Post Graduate Committee in Veterinary Science”, pp. 77-87.
University of Sydney.
Thompson, R.C.A. (1994). Biology and systematics of Echinococcus. In
“Echinococcus and Hydatid Disease” (R.C.A. Thompson and A.J. Lymbery,

eds). CAB International, Wallingford (in press).
Thompson, R.C.A. and Allsopp, C.E. (1988). “Hydatidosis: Veterinary Perspectives
and Annotated Bibliography”. Commonwealth Agricultural Bureau, Wallingford.
Thompson, R.C.A. and Lymbery, A.J. (1988). The nature, extent and significance
of variation within the genus Echinococcus. Advances in Parasitology 27,
210-258.
Thompson, R.C.A. and Lymbery, A.J. (1990a). Echinococcus: biology and strain
variation. International Journal for Parasitology 20, 457470.
Thompson, R.C.A. and Lymbery, A.J. (1990b). Intraspecific variation in parasites
- what is a strain? Parasitology Today 6, 345-348.
Thompson, R.C.A. and Lymbery, A.J. (1991). The epidemiological significance of
biological variation in Echinococcus. Archivos De La Hidatidosis 30, 195-200.
Thompson, R.C.A. and Smyth, J.D. (1975). Equine hydatidosis: a review of the
current status in Great Britain and the results of an epidemiological survey.
Thompson, R.C.A., Kumaratilake, L.M. and Eckert, J. (1984). Observations on
Echinococcus granulosus of cattle origin in Switzerland. International Journal
Thorpe, J.T. (1982). The molecular clock hypothesis: biochemical evolution,
genetic differentiation and systematics. Annual Review of Ecology and Systema-
tics 13, 139-168.
Thorpe, J.P. (1983), Enzyme variation, genetic distance and evolutionary diver-
gence in relation to levels of taxonomic separation. In “Protein Polymorphism:
Adaptive and Taxonomic Significance: (G.S. Oxford and D. Rollinson, eds), pp.
131-152. Academic Press, London.
Verster, A.J.M. (1965). Review of Echinococcus species in South Africa. Onder-
stepoort Journal of Veterinary Research 32, 7-1 18.
Vogel, H. (1957). Uber den Echinococcus multilocularis Siiddeutschlands. I. Das
Bandwurm-Stadium von Stammen menschlicher und tierischer Herkunft. Zeit-
schrijl fur Tropenmedizine und Parasitologie 8, 404454.
Vogel, M., Miiller, N.M., Gottstein, B., Flury, K., Eckert, J. and Seebeck, T.
(1991). Echinococcus multilocularis: characterisation of a DNA probe. Acta
Tropica 48, 109-1 16.
Wachira, T.M., Bowles, J., Zeyhle, E. and McManus, D.P. (1993). Molecular
examination of the sympatry and distribution of sheep and camel strains of
Echinococcus granulosus in Kenya. American Journal of Tropical Medicine
and Hygiene 48,473479.
Wiley, E.O. (1978). The evolutionary species concept reconsidered. Systematic
Zoology 27, 17-26.
Wiley, E.O. (1981). “Phylogenetics: The Theory and Practice of Phylogenetic
Systematics”. Wiley, New York.
Williams, R.J. and Sweatman, G.K. (1963). On the transmission, biology and
morphology of Echinococcus granulosus equinus, a new subspecies of hydatid
tapeworm in horses in Great Britain. Parasitology 53, 391-407.
Wilson, J.F., Diddams, A.C. and Rausch, R.L. (1968). Cystic hydatid disease in
Alaska. A review of 101 autochthonous cases of Echinococcus granulosus
infection. American Review of Respiratory Diseases 98, 1-15.
Worbes, H., Thompson, R.C.A. and Eckert, J. (1989). Occurrence of the cattle
strain of Echinococcus granulosus in the German Democratic Republic.
Parasitology Research, 75,495497.
How Schistosomes Profit from the Stress
Responses They Elicit in Their Hosts
Marijke De Jong-Brink
Graduate School of Neurosciences Amsterdam. Research Institute

Neurosciences Vrije Universiteit. Faculty of Biology. Vrije Universiteit. De
Boelelaan 1087. 1081 HV Amsterdam. The Netherlands
.
1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 178
1.1. Problems to be solved by schistosomes in their hosts ........ 178
1.2. Interference with the host's regulatory systems . . . . . . . . . . . 180
1.3. Parasitic components/products as candidates for interference with
regulatory mechanisms in the host ................. 182
1.4. Advantages of the model Trichobilhania ocellata-Lymnaea stagnalis
for studying the effects of schistosomes on the regulatory systems
of their hosts ............................ 190
.
2 The Effects of Trichobilhania ocellata on its Snail Host Lymnaea stagnalis
with Reference to Other Schistosome-Snail Partnerships ........ 190
2.1. Effects on internal defence ..................... 190
2.2. Effects on reproduction . . . . . . . . . . . . . . . . . . . . . . . 201
2.3. Effects on metabolism and growth ................. 202
. .
3 How T ocellata Affects Reproduction and Growth of its Snail Host ... 204
3.1. Peripheral effects . . . . . . . . . . . . . . . . . . . . . . . . . . 204
3.2. Central effects ........................... 212
3.3. Schistosomin: origin and induction of its release by a parasite-derived
factor ................................ 214
. .....................
4 Parasites: long-term stressors? 219
...................
4.1. The stressconcept in mammals 219
. . ................
4.2. T ocellata: a stressor for L stagnalis? 222
4.3. Schistosomes: stressors for their vertebrate hosts? . . . . . . . . . 227
4.4. Non-schistosome parasite-host combinations . . . . . . . . . . . . 228
.
5 Summary and Conclusions ....................... 234
Acknowledgements ........................... 237
References ............................... 238
ADVANCES IN PARASITOLOGY VOL 35 Copyrighl 0 1995 Academic Press Limited

ISBN 0-12-0317354 All rights of reproducfion in any form reserved
178 MARIJKE DE JONG-BRINK
1. INTRODUCTION
1.1. Problems to be Solved by Schistosomes in Their Hosts
The complex life cycle of trematodes comprises at least two hosts, i.e., one
or more intermediate hosts and a definitive host. The parasitic worms that
cause schistosomiasis (bilharzia) in man belong to these trematodes (genus
Schistosoma). Asexual multiplication occurs in the intermediate hosts
(freshwater snails), whereas the parasites reach sexual maturity in the
definitive hosts (man or other mammal$). Eggs laid by the adult female
pass through the wall of the intestine or the bladder and are voided with the
faeces or urine of the host. If the eggs find themselves in water they hatch
and produce a ciliated larva (miracidium), which, for its further develop-
ment, must meet a compatible snail. After penetrating through the skin of
the mantle or the head-foot of the snail, the miracidium transforms near the
site of penetration into a primary (mother) sporocyst in which secondary or
daughter sporocysts develop. The daughter sporocysts leave the mother
sporocyst and migrate to the hind part of the snail, the digestive gland/
ovotestis area, where they grow and give rise to the final larval stage, the
cercariae, the production of which may continue for the rest of the life of
the snail. Upon an appropriate stimulus, e.g., a light stimulus, the cercariae
leave the snail. They have a brief swimming life and must soon enter a
suitable definitive host. After penetration through the skin of the defini-
tive host, the development of the cercariae into adult worms takes 2-3
months. The adults live in blood vessels, the sexes are separate and the
female is carried by the male. The fertilized eggs of the parasite leave the
host. As soon as the miracidium has reached a suitable snail the cycle can
. continue.
The major problems which schistosomes have to overcome during their
life cycle are the following: (1) they have to find their hosts (with which
they are compatible) and to penetrate their skin and tissue (for reviews see
’ Haas and Voigt, 1988; Haas, 1992), (2) they have to adapt to environmental
stress induced by changes of physicochemical factors as light, osmolarity,
pH, Pco2,Poz, and glucose concentration, (3) they have to evade immune
attacks in their hosts and (4) they have to obtain energy and space within
their hosts enabling them to grow and reproduce prior to transmission.
Especially the latter two points are relevant for the stages in the vertebrate
as well as in the invertebrate host and in this review we will focus on these
two points with the aim of unravelling the strategies they employ inside
their hosts in order to survive, to continue their development and to
reproduce.
HOW SCHISTOSOMES PROFIT FROM STRESS RESPONSES ELICITED IN THEIR HOSTS 179
1.1.1. Evasion of Immune Attack

Both in their vertebrate and invertebrate host schistosomes have to evade
activities of the immune system, irrespective of the fact that the immune
system of freshwater snails is rather simple compared to the vertebrate
system. It consists of cytotoxic, macrophage-like phagocytes, but it lacks
lymphocytes, immunoglobulins and specific anamnestic responses. It has
the capacity to discriminate between self and non-self and to eliminate
non-self, e.g., parasites (Van der Knaap and Loker, 1990).
The immunoevasion strategies employed by schistosomes in their hosts
comprise the following morphological and physiological aspects (for
reviews see Wakelin, 1984, 1988; Behnke, 1990; Van der Knaap and
Loker, 1990; Cox and Liew, 1991; Cox, 1993; Maizels et al., 1993):
1. reduction of their own antigenicity in order to avoid induction of a
disastrous immune response by processes as molecular mimicry
(Damian, 1964, 1987), acquisition of host antigens (masking), antigen
shedding and antigenic variation;
2. resistance to host-effectors as oxygen radicals and immunoglobulins
by secretion of detoxifying, anti-oxidant enzymes, by thickening of
their tegument, by relocation in the host and/or by degrading host
immunoglobulins;
3. modulation of immunocompetence of the host by interfering in a
humoral way with specific functions of immunocompetent cells and/or
with the capacity of these cells to communicate with other cells by
signalling molecules, cytokines or cytokine-like molecules.
1.1.2. Acquisition of Energy and Space

After having invaded their host and escaped the initial attack of the
immune system, schistosomes migrate to places with a high concentration
of nutrients: blood vessels of the closed vascular system in their vertebrate
host and connective tissue, bathed in haemolymph of the open circulatory
system, in their invertebrate host. In addition, they show morphological and
physiological adaptations to optimize uptake of nutrients through their
tegument and, in adult worms, also through gut epithelium. This suggests
that they obtain energy by competing with their host for energy-rich and
other essential nutrients. However, there are several indications that
parasites in meeting their energy demands generally affect their host in
a much more complex way than merely by competition for nutrients.
They may change their host’s nutrition, i.e., food intake, digestion, assimi-
lation and metabolic conversion efficiency, but also nutritionally related,
energy-consuming physiological processes as growth and reproduction
(Hurd, 1990; Thompson, 1990). The extent to which parasites have to
interfere with reproduction and growth in their host depends on several

factors such as the ratio parasite biomass/host size, and the age and the
nutritional status of the-host. This might also explain that the effects of
schistosomes on reproduction and growth may vary greatly and - in
view of the first point - that the effects on their definitive host are less
clear than those on their intermediate host. For some schistosomes it has
been shown that they reduce or completely stop reproduction in their snail
host (“parasitic castration”) and inhibit or - unexpectedly - enhance
body growth (“giant growth”) (reviews, for example, by Hurd, 1990; De
Jong-Brink, 1990, 1992b).
Experiments (Joosse and Van Elk, 1986) have shown that the increased
body growth of Lymnaea sfagnalis infected with the bird schistosome
Trichobilharzia ocellafa does not reflect an increase of the dry weight of
the snails, but of the wet weight. It therefore does not cost much energy.
The importance of this giant growth is that it results in an increase of the
space occupied by haemolymph, i.e., space where the parasites develop and
multiply. This seems particularly relevant because T. ocellafu produces
enormous numbers of cercariae.
The effects on the inversely related processes of reproduction and growth
cannot be explained by assuming that schistosomes and their snail host
compete for nutrients with the parasite as the winner, leaving the host not
enough energy for growth and/or reproduction. Already at an early stage of
infection, when the parasites are still in the mother sporocyst stage, i.e.,
when the parasite’s need for nutrients is still limited, the effects on
reproduction are obvious (Meuleman, 1972; Sluiters, 1981). This strongly
suggests that a more plausible explanation for these effects on the host is
that parasites interfere with the neuroendocrine system (NES)regulating
reproduction and growth in their host (cf., McClelland and Bourns, 1969;
Meuleman, 1972).
1.2. Interference with the Host‘s Regulatory Systems
In view of the foregoing, it would follow that schistosomes are able to

interfere with the action of two regulatory systems in their host: not only
the immune system (IS), in invertebrates preferably called the internal
defence system (IDS), but also the NES. In considering this strategy of
interference with the two regulatory systems it has, furthermore, to be
taken into account that these systems communicate with each other.
Evidence is accumulating that in vertebrates there is bidirectional com-
munication between the two regulatory systems, the neuroendocrine
system and the immune system (Blalock, 1989; Weigent ef al., 1990;
Imura ef al., 1991; Nistic6, 1993). Neuropeptides not only appear to affect
the activity of cells of the immune system, as is reflected by the presence of

neuroendocrine receptors on these cells (Carr, 1992), they are also pro-
duced by them, acting as paracrine agents (Lolait et al., 1986; Smith, 1989;
Smith and Johnson, 1991). The already large number of neuropeptides
which have been demonstrated to have an effect on IS cells of mammals
is still increasing (Johnson et al., 1992). In addition, factors previously
supposed to be exclusively produced by and acting on cells of the immune
system, cytokines (interleukins, tumour necrosis factor, TNF), not only
exert effects on the neuroendocrine system (Bateman et al., 1989; Navarra
et al., 1991), but are also produced by it (Vankelcom et al., 1989; Lechan et
al., 1990; Blalock, 1992).
As far as invertebrates are concerned comparable phenomena have been
reported. Some neuropeptides (e.g., opioids) have appeared to influence
activity of defence cells, the haemocytes, whereas immunocytochemical
data have indicated that some neuropeptides are also synthesized in haemo-
cytes (for review, see for example, De Jong-Brink, 1992a). Fractions with
vertebrate cytokine-like activities have also been identified in a number of
invertebrates (Beck et al., 1989): they showed cross-reactivity with antisera
raised against cytokines derived from vertebrates and were effective in
assays using mammalian immune cells (Hughes et al., 1990; Beck and
Habicht, 1991a; Raftos et al., 1991). Molluscan haemocytes might produce
these cytokines as they showed immunostaining with antisera raised
against cytokines from vertebrates (Ottaviani et al., 1993b). This indicates
that they have been conserved during evolution. The factor with interleukin
I (IL I) activity isolated from the echinoderm Asterias forbesi is partially
identical to that of mammalian IL I (Beck and Habicht, 1991b). Further-
more, vertebrate-derived cytokines affect the activity of molluscan haemo-
cytes (Stefan0 et al., 1991b). However, it is as yet unknown whether in
invertebrates too these cytokine-like molecules are produced by cells of
the neuroendocrine system and/or have effect on their activity. Never-
theless, the strong similarity between the situation in vertebrates and
invertebrates seems to suggest that these questions will also be answered
in the affirmative for invertebrates in the near future.
These findings on the communication between the neuroendocrine and
the immune system are not only illustrative of the rapid progress of the
field of “neuro-immunoendocrinology”, but also have consequences for
the study of the mechanisms which underly the physiological changes
induced by parasites in their host. Changes at the level of the neuroendo-
crine regulatory system have to be considered in relation to (effects on) the
activities of the immunehnternal defence system. This requires extensive
knowledge of both systems in the host. The picture which is already
complicated if the effects on both systems are considered separately,
becomes even more confusing and difficult to unravel when the effects
are studied in connection with each other. This might explain why studies
available on this topic with respect to parasitic infections are rather limited
and fragmentary.
1.3. Parasitic Components/Products as Candidates for Interference

with Regulatory Mechanisms in the Host
Apart from mechanical damage, interference of schistosomes with regula-

tory mechanisms in their host has to be mediated by parasite components
of the parasite-host interface exposed to elements of these regulatory
systems or by products secreted into the host. Therefore, it is worthwhile
to describe first these components/products and their possible role in the
schistosome-host partnership.
1.3.1. Tegument and Sugace Coat Components

The outer surface of schistosomes is constituted by a tegument, a syncytial
cytoplasmic layer. This tegument with its surface coat, the carbohydrate-
rich components of the plasma membrane, protects them against mechanical
forces and chemical attack by (products of) the host. Nutrients from the
host are taken up via this outer layer and in adult worms, after enzyme
cleavage, also through the gut epithelium.
The surface coat plays a crucial role both in eliciting immune responses
of the host and in immunoevasion. Its importance is reflected by its
dynamic nature: thickness and composition change considerably during
the parasite’s life cycle. Variation in the carbohydrate and peptide surface
epitope expression of schistosomula of Schistosoma mansoni is correlated
to the age - and hence to the localization - of the schistosomula in the
host (Langley and Dunne, 1992). Schistosomes may also acquire host
molecules and anchor them to their own surface. An example is the
presence at their surface of a regulatory protein of the host, the decay
accelerating factor, which controls activation of the alternative comple-
ment pathway in the host (Pearce et al., 1990; Horta and Ramalho-Pinto,
1991).
The molecules of the surface coat not only exert their effects in the host
when they are inserted into or attached to the plasma membrane (surface
associated antigens), but also when they become detached by enzymatic
cleavage and circulate in the host (circulating antigens; Simpson and
Smithers, 1985).
The extracellular domain of receptors, which is localized in the surface
coat too, as, for example, that of the epidermal growth factor receptor in
cercariae and adult schistosomes (Shoemaker et al., 1992), might also be
removed by enzyme cleavage and become soluble. As these soluble

receptors retain their binding properties, they may function as inhibitors
of host molecules, as is the case with soluble forms of receptors for several
cytokines and for hormones in vertebrates (hosts) (Roitt et al., 1993).
1.3.2. ExcretorylSecretory Products

Products released by parasites are often referred to as excretory/secretory
(E/S) products because products of both excretion and secretion are
involved (Stirewalt, 1974; Lightowlers and Richard, 1988).
(a) Excretory products. Excretory products, intermediate and end pro-
ducts of metabolic and catabolic processes in parasites, are released into
the host. Parasitic helminths may excrete lactic, succinic, acetic, propionic
and other fatty acids. As acid excretion in helminths may also change
during development, it is supposed that these excretion products are
advantageous for the parasite and/or for the host (for discussion see
Bryant, 1993). It is known that some parasites excrete their amino nitrogen
-in addition to ammonia - as amino acids. Some amino acids, especially
neutral ones, are known to have electrogenic effects on neurons (Kehoe,
1975). For that reason a release of amino acids by parasites in the host
might have an effect on the neuroendocrine system. The same could be
the case with other as yet unrecognized metabolic intermediates or end
products.
(b) Secretory products. Secretory products, which are released from
glandular cells or tegumental cells of schistosomes (e.g., Meuleman et
al., 1978; Dunn and Yoshino, 1988) by means of exocytosis as well as
diffusible, regulatory molecules (as, for example, steroids) from variable
origin, are also candidates for interfering with the regulatory systems in the
host.
(i) Hormone-like substances. A role in interference with the neuroendo-
crine system and/or the immune system of the host is self-evident if
secretory material, either directly released into the host or transitionally
attached to the surface coat of the parasite, is identical to or strongly
resembles peptide hormones or other regulatory signalling substances in
the host. For example, the secretory material in (sub)tegumental cells of
several plathyhelminths has appeared to react with antisera raised against
neuropeptides derived from vertebrates (Solis Soto and De Jong-Brink,
1994). Although the functional significance of these observations is not
yet clear, the data obtained with cercariae of S . mansoni might give some
indication. Their tegumental cells appeared to show immunoreactivity with
an antiserum to somatostatin. As secretion of this material has been
observed after invasion of the vertebrate host (McLaren, 1980), it might
be functional in the definitive host supposedly in immunosuppression as
has been demonstrated for somatostatin (cf., Payan et al., 1984; Eglezos et
al., 1993).
(ii) Enzymes. Schistosomes secrete a variety of proteolytic enzymes,
proteases, in the different stages of their life cycle. The presence of large
glands or glandular cells where these enzymes are produced, for example,
the escape and acetabular glands in cercariae, and the occurrence of
surface-associated proteases indicate an obvious role in parasite-host
interactions. They function not only in invasion of host tissue, in transfor-
mation and in nutrition but also in immunoevasion (e.g., by degrading host
immunoglobulins) (for review see McKerrow and Doenhoff, 1988). As
mother sporocysts in vitro continue to: secrete a low level of active
. cysteine proteinase Yoshino et al. (1993) suppose that these proteinases
may also play a role in the establishment/maintenanceof infections within
the snail host. It seems also a powerful mechanism to interfere with
regulatory systems if parasitic enzymes would be able to detach receptors
from cells of the host (cf., De Carvalho et al., 1993). Their antigenic nature
has made them candidates for serodiagnosis and immunoprophylaxis.
The same applies to detoxifying or anti-oxidant enzymes, which are
produced and secreted by schistosomes as a response to immunological
stress caused by oxygen radicals, nitric oxide and other toxic molecules
released by cells of the host’s immune systedinternal defence system (for
review see Adema et al., 1991a; Brophy and Pritchard, 1992). These
molecules can attack parasite proteins, nucleic acids and membrane
lipids. In the absence of detoxifying enzymes this may result in parasite
killing (cf., Liew and Cox, 1991). An anti-oxidant substance has been
detected in E/S products of S. mansoni sporocysts (Connors et al., 1991).
For schistosomula and adult worms glutathione transferases (GSTs), gluta-
thione peroxidase (GPx) and superoxide dismutases (SODs) have been
reported to play an important role in defence. Because these cytosolic
enzymes are secreted and - transitionally - bound to the surface they
are important candidate vaccine antigens. Much attention has been paid to
their molecular cloning and sequencing (GSTs, Balloul et al., 1987;
Mitchell, 1989; Nare et al., 1991, 1992; Trottein et al., 1992; G h ,
Williams et al., 1991; SODs, Hong et al., 1992, 1993). For S. mansoni, it
has been demonstrated that the activity of these membrane-associated
enzymes increases significantly with the maturation of the worms. This
increase in activity was found to be positively correlated with the resistance
to oxidants (Callahan et al., 1988; Nare et al., 1990). The fact that inter-
species variation exists, as has been shown for the 28 kDa GSTs (Trottein
et al., 1992), might indicate that they play a role in the compatibility
between parasite and host.
Surprisingly, many of the circulating schistosome antigens that have
been identified are cytosolic metabolic enzymes including at least two
glycolytic enzymes, triose phosphate isomerase (Ham et al., 1992) and

glyceraldehyde 3-phosphate dehydrogenase (Goudot-Crozei’ et al., 1989).
The basis of the protective immunity that these proteins induce is not well
understood, as hexokinase for instance, the rate-limiting enzyme in schisto-
soma1 glycolysis, was shown to be poorly immunogenic (Shoemaker et al.,
1994).
(iii) Heat shock proteins (HSPs). In the context of this review important
examples of proteins in the surface coat of schistosomes and other parasites
are heat shock proteins (HSPs; Maresca and Carat& 1992). HSPs, which
appear to occur in all living cells, are classified in families according to
their molecular weight. Members of the HSP 60 and 70 families function as
molecular chaperones: they interact with unfolded peptides facilitating their
intracellular transport and preventing premature aggregation (Welch, 1993).
They also have an impact on other biological events involving dynamic
changes in macromolecular assembly/disassembly events such as antigen
presentation and exo- and endocytosis (Georgopoulos and Welch, 1993).
Traditionally they were supposed to be located only intracellularly and to
become upregulated by high temperatures in order to protect cells from the
disruptive effects of denaturated peptides. However, their function is not
restricted to the protection against an increase in temperature but also
against a variety of other adverse conditions such as low temperature
(Joplin et al., 1990) and osmotic shock. Therefore these HSPs are also
called “stress proteins”. The distinct members of the HSP families show
differences in their subcellular localization (cytoplasm, nucleus, ER, Golgi
apparatus, mitochondria) related to the processes they facilitate (Welch ef
al., 1991). Later studies, however, have shown that they can also be found
at the cell surface and that they are actively released by parasites (Emani
and Teale, 1993).
The questions of how these HSPs are sorted to the cell surface and are
anchored to it have not yet been solved. One of the mechanisms involved in
transport across the cell membrane might be the pathway proposed by
Kuchler ( 1993). Transmembrane translocators or “chaperone” molecules
export polypeptides by a route independent of the typical secretory path-
way. The substrates or molecules to be transported by these translocators
need to be slightly hydrophobic or conjugated to a lipophilic substituent.
Such a mechanism might also be involved in the release of other molecules
lacking a signal peptide, for example, I1 1a and I1 1p and fibroblast growth
factors. A similar transport mechanism for protein trafficking from parasite
to host (cell) compartments has been proposed by Lingelbach (1993) for
Plasmodium falciparum proteins also lacking N-terminal signal sequences.
As to anchoring, there are indications obtained by immuno-
electronmicroscopy, that HSPs are, at least partly, bound to the cell surface.
The HSP 70 family, which is the predominant HSP family in schistosomes,
does not have transmembrane domains. It also seems unlikely that they are
linked to the surface by a phosphatidylinositol-glycan linkage because they
do not possess hydrophobic C-terminal sequences (Srivastava and Maki,
1991).
It is unclear how cells recognize environmental changes and the HSP
gene expression is activated (Welch ef al., 1991). For cercariae of S.
mansoni it has been demonstrated that environmental stimuli can only
activate HSP 70 gene expression after the tails have been removed
(Neumann ef af., 1993). It has been speculated that the tails produce
inhibiting signals that diffuse to the bodies and suppress their HSP 70
genes. These data seem to complete those obtained by Tielens ef al.
. (1993) who found that neither heat shock nor in vifro transformation
(loss of tails) had any effect on the pattern of protein synthesis - and
hence of HSPs - in sporocysts and/or cercariae of S. mansoni. Apparently,
both heat shock and transformation are necessary for induction of HSP 70
in the cercariae. This conclusion, however, contradicts data obtained by
other authors showing that only a heat shock is sufficient to induce HSP 70
gene expression (Yuckenberg et af., 1987; Blanton and Licate, 1992). The
expression of a HSP 60 homologue in all stages of S. mansoni has been
demonstrated (Tielens et al., 1993). Expression of HSP genes is probably
linked to the expression of other genes involved in parasite differentiation
and development (Polla, 1991).
HSPs, mainly HSP 70, are among the dominant antigens recognized by
the immune system - humoral, cellular, or both - in a large spectrum of
parasites (Kaufmann, 1990a, b; Young ef af., 1990; Estes and Teale, 1991;
Winfield and Jarjour, 1991). The fact that HSPs interact with mammalian
T cells can be ascribed to their structural and functional features which
assures that they are efficiently processed and presented at the macrophage
surface resulting in an interaction with T cells (Shinnick, 1991).
Proteins of the HSP 90 family of some parasites, among them S.
mansoni, have also been reported to be antigenic (Shinnick, 1991). The
members of this family play an important role in the prevention of steroid
receptor binding to DNA and of the phosphorylation of tyrosine kinase in
the absence of the proper stimulus. They keep the receptors inactive until
the signal for activation is received (Lindquist and Craig, 1988). This
seems very important in the refractory period after a stress stimulus.
Comparison between HSPs from different organisms has revealed that
they have been highly conserved during evolution. For several pathogens
the immune response to the HSPs is directed predominantly towards spe-
cific, non-conserved epitopes (Shinnick, 1991) and these non-conserved
epitopes might serve as an “immunological smoke screen” to divert the
host’s immune response from the conserved epitopes, which may be the
regions required for functional activity. The slightly different explanation
given by Kaufmann (1990b) is that the cross-reactive response to broadly

distributed epitopes of HSPs could provide the host with a first line of
defence that can be rapidly called into action prior to activation of species-
specific immunity, which becomes predominant. In both options, the
appearance/presence of these HSPs at the surface of parasites might
represint an important survival strategy of parasites (Newport et al.,
1988; Kaufmann, 1990b; Shinnick, 1991).
(iv) Diffusible molecules. Parasites also have components which can,
either by simple diffusion or by facilitated diffusion, pass the cell mem-
brane into the host. Examples are the regulatory compounds steroids,
ecdysteroids, JH-like components, prostaglandins, and other lipophilic
molecules.
Vertebrate-type steroids like androgens, prostagens and oestrogens have
been demonstrated in a wide range of invertebrate species, including para-
sitic helminths, non-parasitic nematodes, molluscs and insects (Schallig et
al., 1992a). Although steroid transformation has been demonstrated in
invertebrates, de novo synthesis of these vertebrate types of steroids in
invertebrates is still questionable. The same holds true for their function. It
has been suggested that these steroids might play a role in the sexual
development of helminth parasites but direct evidence for a hormonal
role is still lacking (Morrison et al., 1986). They might also be involved
in parasite-host interactions. During infection with T. ocellata, for example,
an increase in the relative concentration of steroids, particularly of andro-
gens, has been observed in the haemolymph of L. stagnalis, suggesting
release by parasites into the host (Joosse, 1984). If so, the question would
be what the normal function is of steroids in the host and what would be
the effect of elevation. In gonochoristic prosobranch snails, it has recently
been shown that testosterone is involved in the differentiation of male
characteristics (Oehlmann and Bettin, 1992).
The role of steroids in the internal defence of the snail host has not been
investigated. This might be interesting because they have appeared to play
an important immunological role in vertebrates during infection. The male
sex hormone testosterone is known to enhance immunosuppression. This
leads to an increase in susceptibility with age of male vertebrates to a wide
variety of infections (Alexander and Stimpson, 1988; Bundy, 1988; Brabin
and Brabin, 1992). Androgens tend to suppress both humoral- and cell-
mediated responses, whereas oestrogens seem to enhance humoral
responses. That both androgens and oestrogens affect the immune response
directly is reflected by the presence of receptors for both sex steroids on
leucocytes and lymphoid tissue (see Alexander and Stimpson, 1988).
In view of these findings on the functions of steroids in the host, a role
for parasite-derived steroids in interaction with the regulatory systems in
the host might be possible.
Figure 1 Light micrograph of a section of the digestive gland-gonad area of

Lymnaea stagnalis parasitized with Trichobilharzia ocellata immunostained with a
polyclonal antiserum against an ecdysone-carboxymethoxin-BSA conjugate.
Immunopositive parenchymal cells are present in the tail (t) of the cercaria. b,
body part of a cercaria. (From De Jong-Brink et al., 1989).
The occurrence of ecdysteroids is not confined to insects and nematodes.

They have also been detected and identified in several other invertebrate
phyla belonging to the Protostomia, including schistosomes (Figure 1;
De Jong-Brink et al., 1989; Schallig et al., 1991a). They have attracted
the attention of many investigators because schistosome infections in
mammalian hosts seemed to lead to detectable amounts of ecdysteroids,
which could be used to diagnose the disease (NirdC et af., 1984). However,
later studies have led to the conclusion that ecdysteroids in parasitized
mammalian hosts should be regarded merely as ecdysteroid-like immuno-
reactive compounds originating from the food (Koolman, 1990). Although
their source and function in schistosomes is not clear they are supposed to
be involved in development and/or reproduction of the parasite. In addi-
tion, parasite ecdysteroids might have an immunosuppressive effect on the
host’s immune response (Barker et a f . , 1990). Another possibility is that
they influence neurons and neurosecretory cells of the host as can be
concluded from the immunocytochemical data showing receptors for
ecdysteroids on these cells within the CNS of an insect (Calliphoru
vicina; Bidmon and Koolman, 1989). Such an influence of ecdysteroids
on neuroendocrine cells might also explain their enhancing effects on
growth and egg production in Biomphalaria glabrata (Shiff and Dossaji,

1991).
Juvenile hormones (JHs) with a dihomosesquita prenoid skeleton, may
also be released by parasites into their host. Insects and crustaceans are the
only taxonomic groups in which the synthesis of JHs has been clearly
demonstrated, Compounds showing JH activity in insect bioassays can
also be extracted from tissues of vertebrates, of many other invertebrate
taxa and even from plants and bacteria (Davey, 1988). However, informa-
tion on the occurrence and synthesis of JH-like materials in non-arthropods
is fragmentary and contradictory. Recently the biosynthesis of isoprenoid
compounds has been demonstrated in adult S. mansoni (Fosterer al., 1993).
The short-chain isoprenoid alcohols included farnesol, which is the pre-
cursor of juvenile hormones in insects. Nothing is known with certainty
about its function in non-arthropods. When JH derived from insects is
applied to nematodes, it has an effect on the control of ecdysis and
hatching, although very large doses are usually required. Therefore, Davey
(1988) concludes that “at this stage JH should be regarded as a useful
tool with which to probe the interaction between the environment and the
endocrine system of nematodes, rather than as a component of this
system”. In view of the limited indications for a biological role of JHs
in non-arthropod hosts in general, a role in schistosome-host interactions
seems doubtful.
Eicosanoids which are arachidonic acid metabolites, are synthesized
almost universally in eukaryotic cells in response to external stimuli.
They play a regulatory role as micro-environmental hormones in several
physiological processes in parasitic infections. Parasitic eicosanoids have
extracellular, short-range effects on cells of the host.
After being exposed to linoleate of the host’s skin, prostaglandins (PGs)
and eicosanoids in the leukotriene class were produced by cercariae in
detectable amounts. It has been shown that they promote cercarial penetra-
tion and transformation and act as vasodilators in the host (Salafsky and
Fusco, 1987; Fusco et al., 1993). In addition, some of these eicosanoids
serve as immunomodulators aiding the parasites to evade the host’s
immune attack (Fukiishima et al., 1993). The data obtained with cercariae
of T. ocellata and S . mansoni support the idea that eicosanoids are involved
in immunoevasion. The eicosanoids produced by these two types of
cercariae in the presence of linoleic acid appeared to inhibit the super-
oxide production by human neutrophils (Nevhutalic et al., 1993). However,
an explanation for the fact that the cercariae of these two parasite species,
which clearly differ with respect to immuno-evasion in the same host
species, synthesize the same types of eicosanoids in similar quantities
might be that eicosanoids play a general non-specific role in immuno-
suppression. Other factors may determine the specificity of the response,
for example, the antigenic HSPs, which are synthesized upon eicosanoid
stimulation (Amici and Santoro, 1991).
1.4. Advantages of the Model Trichobilharzia ocellata-Lymnaea

stagnalis for Studying the Effects of Schistosomes on the
Regulatory Systems of Their Hosts
For several reasons the combination T. ocellatn-L. stagnalis is very con-

venient for studying the mechanisms underlying the effects of schistosome
parasites on the internal defence system, the neuroendocrine system and on
the possible interaction between these two:systems in their snail host. The
. large parasite biomass produced in this parasite-host combination may
account for the severe effects on reproduction and growth of the host.
The morphology, physiology and neuroendocrinology of L. stagnalis, the
largest freshwater snail, has been studied extensively (for reviews see
Geraerts et al., 1988, 1991; De Jong-Brink, 1990). This facilitates the study
of changes at the level of the neuroendocrine system which underlie the
physiological changes in the snail host caused by the parasite. In addition,
several bioassays are available to detect parasite-induced modifications.
Moreover, haemolymph can easily be obtained from this snail species
and the internal defence system of this snail host has been studied in
detail (Sminia, 1972; Sminia and Van der Knaap, 1981, 1986). The
haemocyte, either circulating or residing in the connective tissue, is the
primary cell type involved in the internal defence. It is capable of phago-
cytosis, encapsulation and killing (Van der Knaap and Loker, 1990). The
development of T. ocellata in the snail has been studied in detail and shows
a great similarity to that of species which cause schistosomiasis (Meuleman
and Holzmann, 1975; Meuleman et al., 1978, 1980; Sluiters et al., 1980;
Sluiters, 1981), which are more important from medical and economical
points of view. The life cycle of T. ocellutu, with ducks as definitive hosts,
can easily be maintained in the laboratory (Meuleman et al., 1984). These
endoparasites have no host tissue eating redial stage, so apart from exerting
mechanical pressure they only interact with the host in a humoral way.
2. THE EFFECTS OF TRlCHOBlLHARZlA OCELLATA ON ITS SNAIL

HOST LYMNAEA STAGNALlS WITH REFERENCE TO OTHER
SCHISTOSOME-SNAIL PARTNERSHIPS
2.1. Effects on Internal Defence
Compatible schistosome parasites may avoid the confrontation with the

defence system of their snail host by molecular disguise: masking of
surface antigens by uptake of snail determinants or mimicry, the production

of surface coat molecules resembling host antigens (for reviews, Bayne and
Yoshino, 1989; Van der Knaap and Loker, 1990). Another strategy used by
schistosomes to circumvent the snail’s defence activities is to interfere with
these activities leading to suppression reflected by changes in numbers,
morphology and/or activity of the haemocytes, the cellular component of
the defence system, which is essential in antitrematode activities (Lie,
1982; Loker and Bayne, 1986; Lie et al., 1987).
Activity of the freely moving haemocytes comprises the following
aspects: they are able to recognize non-self, to synthesize lectins (Van
der Knaap et al., 1981, 1983), carbohydrate binding glycoproteins acting as
agglutinins and opsonins, and other unknown humoral factors, to encapsu-
late (Sminia et al., 1974), to phagocytose and to exert various cytotoxic
reactions. Haemocyte-mediated cytotoxicity (Bayne et al., 1980a, b) is
responsible for killing and digestion of phagocytosed small organisms
and for encapsulation and killing of larger organisms. Both non-oxidative
and oxidative killing mechanisms are involved. Non-oxidative killing is
achieved by the release of factors, produced by the haemocytes, into
phagosomes or into the haemolymph. These factors include lysosomal
enzymes and bactericidins (Cushing et al., 1971; Adema et al., 1991a).
Oxidative killing is achieved by reactive oxygen intermediates (ROIs) and
possibly nitric oxide as described by Dikkeboom et al. (1987) and Adema
et al. (1991b, 1992). These are generated when haemocytes make contact
with non-self or damaged/effete self. Release of ROIs plays a role in
extracellular larvicidal activity and in intracellular killing of phagocytosed
organisms.
In view of their central role in defence it is clear why haemocytes are
the targets of parasite-mediated interference with the host’s internal
defence system. The presence in snails of trematode larval stages or
their E/S products has been shown to alter a variety of haemocyte
functions. This includes cell adherence (Noda and Loker, 1989), lysosomal
enzyme activity (Granath and Yoshino, 1983), polypeptide synthesis and
release (Yoshino and Lodes, 1988; Lodes et al., 1991) and superoxide
production (Connors and Yoshino, 1990). Motility of haemocytes was
shown to be modulated by Lodes and Yoshino (1990), who demonstrated
that several E/S peptides from S. munsoni mother sporocysts diminish
transmembrane migration of haemocytes from B. glabrata. Modulation
of phagocytic activity of haemocytes has been found by many workers
(Abdul Salam and Michelson, 1980a, b; Van der Knaap et al., 1987; Noda
and Loker, 1989; Connors and Yoshino, 1990; Fryer and Bayne, 1990). The
important role of E/S products of parasites in interference with antitrema-
tode cytotoxicity has been demonstrated by experiments in vifro, in which
haemocytes from a B. glabrata strain, which is resistant to S. munsoni and
susceptible to Echinostoma paraensei, killed fewer S . mansoni sporocysts

when the snails had been infected with E . paruensei than did haemocytes
from non-infected B. glabrata (Loker et al., 1986; cf., Lie, 1982). More
recently, it has been detected that haemocytes of this B. glabrata strain lose
their ability to damage S . mansoni sporocysts in vitro when they are
pretreated with E. paraensei E/S products (Loker et al., 1992).
Plasma factors synthesized by haemocytes and also by other snail tissue/
organs (Sminia and Van der Knaap, 1986) play a facilitating role in
antitrematode haemocyte activity (Loker and Bayne, 1982, 1986; Granath
and Yoshino, 1984). This is also illustrated in that parasites induce sub-
stantial changes in soluble haemolymph carbohydrate binding polypeptides
(lectins). By using two strains of B. glabratu, one susceptible (M line) and
the other resistant (10-R2) to S. mansoni and both susceptible to E .
paraensei, it became clear that the two different parasites elicited different
responses in the same host strain and that the two host strains responded
differently to the same parasite with respect to the occurrence of proteins
with agglutinating activity in the haemolymph, polypeptide bands around
8&120 kDa and - less consistently - around 200 kDa (Couch et al.,
1990; Monroy and Loker, 1993).
Two assays have been used for the study of the effects of T. ocellata on
the internal defence of L. stagnafis. In the first in vitro assay phagocytosis
by haemocytes was used as parameter for defence activity (Amen et al.,
Figure 2 Light micrograph of a monolayer of haemocytes from Lymnaea stag-

nalis showing spreading (s) and non-spreading (ns) cells. Some of the spreading
cells are phagocytosing zymosan particles (arrows). (From De Jong-Brink, 1992a.)
1992). In this phagocytosis assay (PA) monolayers of haemocytes were

prepared and allowed to phagocytose zymosan particles (Figure 2). The
percentage of cells having phagocytosed these particles was determined for
individual snails. In the second assay, the bacterial clearance assay (BCA),
changes in the capacity of haemocytes to eliminate bacteria, either extra-
cellularly or intracellularly after having phagocytosed these bacteria, are
used as a parameter for modulation of haemocyte activity (Nuiiez et al.,
1994a). As bacterial target a marine bacterium, Aeromonas salmonicida,
was used which is effectively recognized and phagocytosed by haemocytes
of L. stagnalis. The reduced number of viable bacteria can be quantified by
adding 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide
(MTT),which is reduced by viable bacteria, reading the absorbance on a
multiscan spectrometer and comparing these values with controls, i.e.,
those lacking haemocytes.
The period immediately after penetration of T. ocellata into the snail
host (1.5-72 h post-exposure; pe) was extensively studied with the two
assays because in this period modulation can be expected. In addition
several time points coinciding with the later - successive - stages of
parasite development were investigated with the PA. In all these investiga-
tions a distinction was made between plasma- and cell-associated effects.
Furthermore, E/S products released in vitro by miracidia during their
transformation into mother sporocysts and shortly thereafter, which are
involved in this humoral interference with the host’s IDS, have been
characterized. Complete identification of the factors involved might reveal
the molecular mechanisms employed by schistosomes to modulate the
internal defence in the snail host.
2.1.1. Early Effects

The data obtained with the two assays for defence activity gave essentially
the same results for the period 1.5-6 h post-exposure in vivo, viz. a cell-
as well as a plasma-associated enhancement of activity in parasitized
snails. In the period 12-72 h post-exposure both assays showed a
plasma-associated suppression of activity. A cell-associated suppression
in this period was only detected with the BCA indicating that this assay is
more sensitive (Figure 3). Data obtained with the BCA showed that at
96 h post-exposure no difference was found between haemocyte- and
plasma-associated defence activities of infected and non-infected snails.
An interesting aspect is that suppression of the host’s internal defence is
preceded by a short period of activation. In this period the increased
activity is probably directed against the ciliated plates, shed by the para-
sites directly upon invasion of the snail host. An explanation for the fact
that the newly formed mother sporocyst is not attacked by these activated
0.60 -
0.55 .
+ + + + + +
m m m m m m
Figure 3 Effect of Trichobilharzia ocellata infections on activity of haemocytes

from juvenile Lymnaea stagnalis measured in the bacterial clearance assay. This
assay uses the reduction by bacteria of a tetrazolium dye (MTT) to detect colori-
metrically the number of surviving bacteria. MTT reduction by surviving bacteria
following incubation with heamocytes (H) from non-infected juveniles or from
juvenile snails infected with T. ocellata at 1.5, 24, 48,72 and 96 h post-infection
(PI). MTT reduction is expressed as mean absorbance/well 2SE (n = 12; each
well contains haemocytes from one individual snail). Bacteria with haemocytes
from non-infected juveniles [B + H(NI)] serve as control; other controls are not
presented. (From Nliiiez et al., 1994a.)
haemocytes might be that the parasites also have a suppressive effect on

haemocytes, exerted by a substance, which acts more locally, i.e., carries
only a short distance from the parasite and/or which is overruled systemati-
cally by the effects of a strong activating factor. This explanation is
supported by the results of in vifro experiments in which parasites were
cultured for 96 h and the media were changed after 33 and 72 h. Separation
techniques - gel permeation and HPLC - applied to the culture super-
natants and testing of the fractions in the BCA revealed the presence of an
activating factor and a suppressive factor in all three periods studied
(Figures 4 and 5). In the period up to 33 h (corresponding to ca 6 h in
vivo) the low-molecular-weight (ca 2 kDa) activating fraction predomi-
nated, in the period 33-72 h (corresponding to ca 12-72 h in vivo) the
high-molecular-weight (ca 40 kDa) suppressing factor and in the period
72-96 h both fractions occurred at lower levels and in approximately the
I
0.1:
11
?
MSM 0-33 h r ~
\
sc'9
33-72 h r ~
n l
72-96 hrs
Figure 4 Gel-permeation HPLC profiles of E/S products released by a single

batch of Trichobilharzia ocellata during in vitro transformation of miracidia in
mother sporocysts. E/S products were obtained after 33 h, from 33-72 h and
72-96 h of culturing. Black arrow, 2 kDa fraction; open arrow, 40 kDa fraction.
(Nliiiez et al., 1994b.)
same amounts (Ndiiez et al., 1994b). The suggestion that the parasites
produce two main fractions which together gave a resultant effect on
haemocyte activity was confirmed by testing the combined E/S fractions
of the three time groups of parasite cultures in the BCA. The combined
E/S fractions from 0-33 h cultures gave a resultant effect of increasing
the killing activity of haemocytes, whereas the combined fractions from
33-72 h cultures lowered the killing activity. The E/S fractions from
72-96 h cultures had no significant effect on haemocyte activity. So, the
results obtained in vitro seem to reflect the in vivo situation.
2.1.2. Compatibility Factor?

The observations on the early effects suggest that the suppressive E/S
factor from the parasites acting directly on haemocytes protects the para-
site from being attacked by the host defence system in a compatible
situation. This supposition was investigated by testing its effect on haemo-
cyte activity in an incompatible host, Planorbarius corneus (Ndiiez and De
Jong-Brink, 1994; this species was chosen because the number of circulat-
ing haemocytes in specimens of B. glabrata, the host of S. mansoni also
196
::; 40kD
r c N r n r t N r n
9+ $ +
m E l a l a E H l a
T
$ 4 4
MARIJKE DE JONG-BRINK
MSM MSM+P
Figure 5 Effect of E/S factors produced during in vitro transformation of
Trichobilharzia ocellata miracidia (0-33 h), on the bacterial killing activity of
haemocytes from non-infected juvenile Lymnaea stagnalis. MTT reduction by
surviving bacteria following incubation with haemocytes (H)which had been
pre-incubated with the fractions 1-3 obtained from MSM in which T. ocellata
had been cultured (MSM + P) or the corresponding fractions obtained from MSM
alone. MTT reduction is expressed as mean absorbance/well 2SE (n = 12; each
well contains haemocytes from one individual snail). Incubation of bacteria (B)
only serves as control. (Ndiiez et al., 1994b.)
used in this study, is too low for the BCA). Incubation of haemocytes and
bacteria with the activating factor, isolated from the medium in which T.
ocellata had been cultured, resulted in the haemocytes from both snail
species having an increased activity compared to that of the corresponding
haemocytes incubated in the absence of any factor. The suppressing factor
also had a modulating effect on haemocytes from L. stagnalis. However,
this effect was not observed with haemocytes from P. corneus (Figure 6).
The supposition that this direct suppressive factor of ca 40 kDa is impor-
tant with respect to compatibility between parasite and host is, furthermore,
supported by data obtained in viva In these experiments haemocytes taken
from juvenile L. stagnalis, which had been exposed to or injected with
miracidia of either T, ocellata or S . mansoni were tested for their capacity
to attack bacteria. The activity of haemocytes taken from snails 1.5 h after
HOW SCHISTOSOMES PROFIT FROM STRESS RESPONSES ELfClTED IN THEIR HOSTS 197
0.65
T
0.60
I
0.55
0.50
0.45 7-
3 m
+ 3 m+
m m s p sp m s p
+ + r + + +
L stagllalic P. eomcus
Figure 6 Effect of EIS factors, produced during in vitro transformation of
Trichobilharzia ocellata miracidia, on the bacterial killing activity of haemocytes
from non-infected adult Lymnaea stagnalis or Planorbaris corneus. MTT reduction
by surviving bacteria, at the two concentrations for each of the snail species,
following incubation with haemocytes from L. stagnalis or P. corneus which had
been pre-incubated in the absence or presence of either activating factor (A) or
suppressing factor (S). MTT reduction is expressed as mean absorbancelwell ?SE
(n = 12; each well contains haemocytes from one individual snail). Note: data of
haemocytes with bacteria (H+ B) for each of the snail species are used as controls.
(Ndiiez and De Jong-Brink, 1994.)
being exposed to or injected with either of the two species was significantly
higher than that of cells from the corresponding sham-treated snails.
Haemocytes from snails that had been exposed to or injected with S.
mansoni 24, 48 or 72 h earlier also had higher clearing capacities than
those from the sham-treated snails. This, however, was not the case with
haemocytes from snails exposed to T. ocelluru as these were all suppressed
and hence had lower killing activities at these time points (Figure 7).
In summary, the activating parasitic factor is recognized by haemocytes
of both the compatible and non-compatible host, the parasite-derived
suppressive factor, on the other hand, is not recognized by haemocytes of
non-compatible hosts. This might explain why parasites in an incompatible
combination are encapsulated and eliminated.
These findings corroborate the results obtained with E/S products of 1
day primary cultures of S. munsoni (Yoshino and Lodes, 1988). These E/S
products affect polypeptide synthesis by haemocytes of B. glubrutu.
0 Control
T. o c e W
0.65 Exposure 0.65 Injection S. mansoni
T
0.55 0.55
...
0.45 I 1 I I 0.45
P
e
v!
+ B + H(Nn + B + H(NI)
Figure.7 Effect of Trichobilharzia ocellata or Schistosoma mansoni infections
on the bacterial killing activity of haemocytes from juvenile Lymnaea sfagnalis.
MTT reduction by surviving bacteria following incubation with haemocytes taken
from snails at 1.5, 24, 48 or 96 h post-infection (PI) after either exposure to or
injection with T. ocellata, S. mansoni or S S S (controls). MTT reduction is
expressed as mean absorbance/well 2 S E (n = 12; each well contains haemocytes
from one individual snail). Note: data of bacteria with haemocytes from snails
sham exposed or sham injected are used as controls. (Nliiiez and De Jong-Brink,
1994.)
Haemocytes from a resistant strain showed a greater capacity to be stimu-

lated than those from a susceptible strain. The authors suggest that this
increased polypeptide synthesis in resistant haemocytes by parasite E/S
products might reflect their parasite-killing efficiency. The components of
the E/S products responsible for this modulatory effect were a fraction with
a ,molecular weight >30 kDa and a smaller fraction with a molecular
weight c10 kDa. This latter fraction might be comparable with the activat-
ing fraction of ca 2 kDa in E/S products of T. ocelluru. Inhibition of the
secretion of polypeptides by susceptible haemocytes could be attributed to
a high-molecular-weight protein aggregate comprising subunits of 22-
24 kDa in the E/S products of S. munsoni, whereas the secretion of
resistant haemocytes was not affected. This differential effect was only
observed in the presence of homologous plasma (Lodes er ul., 1991). The
role of plasma components in establishing the effects of T. ocelluru E/S
products on haemocyte activity has not been studied. There is the possibi-
lity that, in the presence of homologous plasma, more fractions would show
modulatory activity, for example, by binding to plasma recognition mole-

cules (lectins). The fact that the resultant effect of the combined E/S
fractions of T. ocellata reflected the in vivo situation renders this possibi-
lity not very likely.
The results obtained for both T. ocellata and S . mansoni clearly show
that the parasite E/S products play a crucial role in phenomena such as
compatibility/susceptibility and incompatibility/resistance.
2.1.3. Later Effects

In addition to these early effects of T. ocellata on the host internal defence
system, the possible defence activity changes in later stages of infection,
namely 2,4, 6, 8 and 10 weeks post-exposure, have also been investigated
(Amen et al., 1992). At these time points the phagocytic activity of
haemocytes from both infected and non-infected snails was determined
in the PA after being incubated with plasma from either infected or non-
infected snails (Figure 8; four combinations at each time point). Only at 8
01
2 4 6 8 10
Time post -exposure (h)
Figure 8 Mean values of the number of phagocytosing haemocytesI500 non-

infected Lymnaea stagnalis or snails infected with Trichobilharzia ocellata (n =
10). The haemocytes were incubated with plasma of infected or non-infected snails
taken at the same time-points, i.e., at 2, 4, 6, 8 and 10 weeks post-exposure.
Significant differences were found at 8 and 10 weeks post-exposure between
haemocytes of infected (inf.) and those of non-infected snails (non-inf.). Plasma
alone had no significant effect. ( 0 )Cells inf.; + plasma inf.; (- -0--) cells non-inf;
+ plasma non-inf.; (- -0- -) cells inf. + plasma non-inf.; (0)cells non-inf. + plasma
inf. (Amen et al., 1992.)
and 10 weeks post-exposure have significant changes been found: a

haemocyte-associated activation. Also in this case the PA is probably not
sensitive enough to detectr plasma-associated effects and effects at other
time points. Moreover, in the PA the resultant of possibly several systemic
effects in vivo is investigated. At 8 and 10 weeks post-exposure the snails
were shedding and during their migration in the snail the cercariae release
non-snail substances and cause lytic and mechanical damage in host
tissues. It seems plausible that the increased activity of haemocytes is
aimed at clearing away non-self and effete self. However, in this situation
it is surprising that the cercariae are not attacked as soon as they leave the
daughter sporocyst. Experiments with a polyclonal antiserum raised against
plasma from the snail host have revealed that as soon as they leave the
'sporocyst, but not before, cercariae are covered with material recognized
by this antiserum (Figure 9). This strongly suggests that masking of their
surface antigens is used as a strategy to escape from the activities of the
host defence system (Van der Knaap et al., 1985). Besides, the possibility
that the observed increase of haemocyte activity is the resultant effect of
several factors in the host, cannot be ruled out.
Figure 9 Light micrograph of a section of the digestive gland (dg)-gonad area of

Lymnaea stagnalis parasitized with Trichobilharzia ocellata immunostained with a
polyclonal antiserum against plasma from patently parasitized snails. Note the
difference between the cercariae outside (black arrows) and inside (open
arrows) the daughter sporocyst.
2.2. Effects on Reproduction
When L. stugnulis is infected soon after hatching -at a size of 2-3 mm -

development of both the male and the female part of the reproductive tract
of this hermaphrodite snail is nearly completely inhibited. However, when
infected at a size of 5-7 mm the development of the male part is less
inhibited than that of the female part in this slightly protandric snail (De
Jong-Brink and Croll, see De Jong-Brink, 1992b). Routinely snails are
infected at a size of 10-11 mm. In these parasitized snails development
of the male and female parts of the reproductive tract is retarded. The effects
of parasitosis on the development of the reproductive tract of Lymnueu can
already be observed within 1-2 weeks post-exposure (Sluiters, 1981).
Oviposition never really starts in these snails infected as juveniles. Similar
-
observations have been described for the combination S. mansoni-B.
glubrutu (e.g., Meier and Meier-Brook, 1981). The observation by Cooper
et al. (1992) that a moderate to high percentage of B. glubrutu, infected as
neonates, was eventually capable of simultaneously producing both eggs
and cercariae is difficult to explain.
In specimens infected at a sub-adult stage, on the other hand, a signifi-
cant increase in fecundity was observed in the first weeks after infection.
401
35
30
25
20
I5
10
0
1 2 3 4 5 6 7 8
weeks post infection
Figure 10 Total number of egg masses produced by snails infected as sub-adults

(m) and by non-infected (0) control snails (n = 25). The black line on top
indicates the presence of schistosomin in haemolymph of infected snails from 5
weeks post-infection onwards, coinciding with the decrease of egg mass production
in these snails. (Modified after Schallig et al., 1991c.)
202 MARUKE DE JONG-BRINK
The increase was followed by a strong decrease in fecundity from 5 weeks

post-exposure onwards (Figure 10; Schallig et al., 1991~).Similar obser-
vations have been described for the combination S. mansoni-B. glabrata
(Thornhill ef al., 1986). No difference in fecundity was found between L.
sfagnalisinfected as adults and non-infected controls during the first four
weeks post-exposure. As with the snails infected, as sub-adults, these
infected snails showed a decrease in fecundity from five weeks post-
exposure onwards. The results obtained with Biomphalaria infected as
adults, on the other hand, are less uniform, and varied from an initial
increase followed by a decrease in fecundity (Minchella and Loverde,
1981) to an almost immediate decrease ,of fecundity (Meuleman, 1972;
Crews and Yoshino, 1989).
2.3. E f f m on Metabolism and Growth
In a number of cases snails infected with schistosomes show an increased

body growth (“giant growth”), such as L. stagnalis infected as juveniles
with T. ocellafa. On the other hand, B. glabrata and B. pfeifferi infected
with S . mansoni showed only a temporary growth acceleration before
patency is reached with the final size being in the normal range (Pan,
1965; Meuleman, 1972; Meier and Meier-Brook, 1981; Thornhill et al.,
1986; Crews and Yoshino, 1989). These differences between infected
Biomphalaria and L. sfagnalismight be related to the lower production of
parasites (cercariae) in B. glabrata compared to that in L. stagnalis (see also
Hurd, 1990). Giant growth of L. sfagnalis,which starts when differentiated
cercariae can be observed in the daughter sporocyst, results in an absolute
and - compared to the digestive gland - relative increase of the space
occupied by haemolymph, i.e., space where the parasites develop and
multiply (Figure 11; De Jong-Brink ef al., 1989). The ratio between the
dry and wet weights of infected L. sfagnalis matches that of non-infected
juveniles, indicating that parasitized snails retain their juvenile body
stpcture (Joosse and Van Elk, 1986). However, the fact that these data
on dry and wet weights include those of the parasites has not been taken
into account.
Parasitic infection also affects the metabolism of infected snails (Kohler
and Voigt, 1988; Thompson, 1990). A decline in the levels of haemolymph
glucose, protein, amino acids and lipids and a depletion of snail tissue
carbohydrates is very conspicuous in schistosome-infectedsnails. Also the
effects of infection on processes associated with metabolic rate as oxygen
consumption, ’feeding actiuity, heart beat, locomotory activity and heat
production have been investigated. B. glabrata snails, infected with S.
mansoni, consume more food (Williams and Gilbertson, 1983) and have
HOW SCHISTOSOMES PROFITFROM STRESS RESPONSES ELICITEDI N THEIR HOSTS 203
Figure 11 Light micrographs of paraffin sections of the digestive gland-gonad

area of a parasitized (1) and a non-parasitized (2) Lymnaea stagnalis. Note the
differences i n relative surface areas of digestive gland (dg) and connective tissue
(c) between the sections. Arrows: daughter sporocysts. (De Jong-Brink et al.,
1989.)
a higher basal metabolic rate than non-infected snails but they reduce their
locomotory activity to compensate and maintain a constant rate of energy
conversion (Becker, 1980). For L. stagnalis it has been shown that food
consumption does not differ in parasitized animals compared to controls,
whereas the assimilation remains initially constant and declines slightly
from the time the daughter sporocysts contain differentiated cercariae and
the snails are shedding (Bayomy et al., 1989). In shedding snails there is a
severe glycogen depletion in the head-foot and mantle region (Joosse,
1988), i.e., from the parts where glycogen-storing vesicular connective
tissue cells are numerous (Sminia, 1972). Although at an earlier stage of
parasitosis, when the daughter sporocysts commence to grow and the
cercariae start to develop, the glucose content in the haemolymph of
infected snails was found to be decreased compared to that of controls,
glycogen depletion was not accompanied or caused by a (second) change in
haemolymph glucose concentration (cf. Becker, 1980). Apparently an, as
yet unknown, control mechanism in the process of glycogen depletion is
involved. In L. stagnalis infected with T . ocellata the haemolymph
protein concentration, mainly haemocyanin, and that of total free amino
204 MARLIKE DE JONG-BRINK
acids (mainly polar amino acids) are lower than in non-infected controls
(Bayomy et al., 1989).
Investigations by Thompson et ul. (1992) have shown that the effects of
nutrient utilization by developing S. mansoni on the snail host may depend
on the snail's diet. The reduction of free phosphatides in the digestive
gland of B. glabrata that coincided with pateicy of' the infection with S.
mnsoni, could not be observed when the snails were maintained on high
fat diets containing egg yolk.
3. HOW T. OCELLATA AFFECTS REPRdDUCTIONAND GROWTH OF

ITSSNAILHOST
To test the hypothesis that T. ocellata affects reproduction and growth of L.

stugnalis by interfering in a humoral way with the neuroendocrine regula-
tion of these processes, the following aspects were studied in the haemo-
lymph of infected snails a humoral factor is present, either derived from the
parasite or, upon stimulation by a parasite-derived factor, from the host
itself, which interferes with the action of snail hormones upon their target
organs (peripheral effects) and/or with synthesis and release of these
hormones (central efects). The hypothesis has been studied in infected
snails approaching or during patency as in this stage of infection the effects
on reproduction and growth are most pronounced.
3.1. Pewiphewal Effects
3.1.1. Mode of Interference with Reproduction

The first aspect has been studied using several bioassays developed for
three female gonadotropic hormones in this hermaphrodite snail. Two of
them, calfluxin (CaFl) and the caudodorsal cell hormone (CDCH), are
neuropeptides derived from the same polypeptide precursor in the neuro-
secretory caudodorsal cells (CDCs) (Vreugdenhil et al., 1988; Geraerts et
al., 1991) of the cerebral ganglia of the CNS (Figures 12 and 13). The third
is the dorsal body hormone (DBH) produced by the endocrine dorsal bodies
(DBs) which are located on the cerebral ganglia (Figure 12). The CDCs
control the process of egg laying and associated behaviours, whereas the
DBs regulate differentiation and growth of the female reproductive tract
and growth &d maturafim of oocytes (Geraerts and Joosse, 1975; De
Jong-Brink and Geraerts, 1982). In the bioassays knowledge on signal
transducing pathways in their target cells was used.
HOW SCHISTOSOMES PROFIT FROM STRESS RESPONSESELICITED IN THEIR HOSTS 205
MDB
LDB
LL
WLNN
Figure 12 Schematic drawing of the right cerebral ganglion of the central
nervous system of L. stagnalis showing the neurosecretory caudodorsal cells
(CDC), the lobus anterior (LA) with the alanine-proline-glycine-tryptophan
(APGW) neurons, the medio- and latero-neurosecretory light green cells (mLGC,
lLGC), the lateral lobe (LL) with the canopy cell (C), which also belongs to the
LGC, and the endocrine medio- and latero-dorsal bodies (MDB, LDB). CC,
cerebral commissure, the neurohaemal area of the CDC; LN, lip nerve, the
neurohaemal area of the LGC.
CaFl stimulates the influx of calcium into mitochondria of the secretory

cells of the albumen gland, one of the female accessory sex glands (Figure
13). The gland secretes perivitellin fluid, a nutritive substance for the
developing embryos, onto fertilized oocytes. The influx of Ca2+is related
to the stimulation of perivitellin fluid release from the secretory cells
by exocytosis. As a measure for the effect of CaFl on glands incubated
with CaFl, the percentage was taken of mitochondria containing Ca-
deposits as demonstrated with the ultracytochemical antimonate precipi-
tation technique. The stimulating effect of CaFl appeared to be inhibited
by a factor, present in haemolymph of infected snails from the time
jl1
% Ca-pos mitochondria
+ 0 -can
+can
20
d
e e
0 0
0
3 6 9 12
weeks pe.
Figure 13 (A)Schematic representation of precursor I of the caudodorsal cell

hormone (CDCH) of Lymnaea showing the localization of calfluxin (CaFl).
(Vreugdenhil et al., 1988.) (B) Electron micrographs of secretory cells of the
albumen gland of Lymnuea sragnalis incubated with (left) or without (right) the
hormone CaFl. Arrows indicate Ca2+deposits in mitochondria (-m-) of the stimu-
lated gland cell: Ca-positive mitochondria. S, secretory granule; rer, rough, endo-
plasmic reticulum. (C) Percentages (means and standard deviations) of Ca-positive
mitochondria in albumen glands of non-infected snails incubated in haemolymph
cercarial differentiation starts within the daughter sporocysts (Figure 13;

De Jong-Brink et af., 1986a, 1988a).
Co-injections of haemolymph from patently infected snails with native
as well as synthetic CDCH, the ovulation inducing hormone, prevented
ovulation (Hordijk et al., 1991a).
DBH enhances adenylate cyclase (AC) and hence CAMP production in
follicle cells in ovotestes of snails that have just ovulated. This activation
probably reflects resumption of oocyte growth and/or inhibition of matura-
tion of remaining, non-ovulated oocytes. An enzyme cytochemical
method is used for the demonstration of AC activity (De Jong-Brink et
al.,, 1986b). The percentage of oocytes surrounded by follicle cells with
lead-imidodiphosphate deposits, indicating AC-activity, is taken as a
measure of the effect of DBH. The effect of DBH on follicle cells was
inhibited when gonads were incubated in haemolymph of snails with patent
infections (De Jong-Brink and Bergamin-Sassen, 1989).
The synthetic activity of the albumen gland, measured as the amount of
[14C] glucose incorporation in its secretion product galactogen, is also
stimulated by DBH (Wijdenes et al., 1983). The increase in synthetic
activity in the presence of DBH was markedly lower in glands incubated
in haemolymph from patently infected snails than in glands incubated in
haemolymph from non-infected snails (Joosse et al., 1988).
So, the bioactivity of all three female gonadotropic hormones studied
appeared to be inhibited in the presence of haemolymph from infected
snails once differentiating cercariae were present in the daughter sporo-
cysts, whereas no inhibition of the hormone response was observed in
haemolymph of snails infected for a short time and in that of non-infected
snails. The inhibitory effect of haemolymph from parasitized snails could
be ascribed to a heat-resistant, pronase-sensitive factor, which was called
schistosomin (De Jong-Brink et al., 1986a, 1988a; Joosse et al., 1988). The
name schistosomin suggests that it is a parasite-derived factor. However,
later experiments have shown that it is a host-derived factor (see below).
Moreover, it was not clear at first whether one or more types of schisto-
somin causing inhibition of different gonadotropic hormones are present in
haemolymph of infected snails.
3.1.2. Mode of Inte$erence with Growth and Metabolism

The picture concerning neuroendocrine mechanisms involved in regulation
of growth and metabolic processes in Lymnaea is not yet complete (De
of respectively 0 , 3 , 6 , 9 or 12 weeks infected snails in the absence (0)

or presence
(m) of CaFl. Each mean is based on counts in five glands (100 mitochondria per
gland). The means per group not sharing a common letter differ significantly.
Comparison between data of incubations -CaR and +CaR is only allowed at
the same time. (De Jong-Brink et al., 1988a.)
208 MARlJKE DE JONG-BRINK
Jong-Brink, 1992b). The neuroendocrine light green cells (LGCs;this term

reflects their staining with Alcian blue/Alcian yellow after previous hydro-
lysis) in the cerebral ganglia of the CNS of L y m e u are involved in
regulation of growth (Figure 12). Each of the two lateral lobes (LL),
which are attached to the cerebral ganglia, contain a large neuron with the
same characteristics as the LGCs, the canopy cells (CCs; Geraerts, 1976).
The LGCs and the CCs synthesize molluscan insulin-related peptides
(MIPs; Smit et ul., 1988). However, the exact functions of the individual
MIPs are unknown. One of the M I P s might be the hyperglycaemic factor
derived from the CNS, which inhibits glycogen synthesis and stimulates
glycogen breakdown and release from: the glycogpn storing vesicular
, connective tissue cells (Sminia, 1972) into the haemolymph (Hemminga
et ul., 1985; Joosse, 1988). For two MIPS(II and V) it has been shown that
their transcript levels decrease quickly after food deprivation (Teunissen,
1994).
Appropriate bioassays are still lacking to study whether the biological
activities of the hormones involved in regulation of body growth and
metabolic processes in Lymnueu are also affected by (a) humoral factor(s)
in haemolymph of parasitized snails. This explains why nothing is know of
the way the parasite interferes with the action of snail hormones involved
in regulation of growth and metabolism. The MIP, which is supposed to
be the hyperglycaemic factor, might be involved in glycogen depletion
in shedding snails. Extirpation of the lateral lobes with the canopy cells,
causes giant growth as does parasitosis (Geraerts 1976, 1992). The
complicated mechanisms underlying giant growth are, however, not
clear.
I
PRIMARY STRUCTUREOFSCHISTWOMIN
*-
Figure 14 Primary structure of the schistosomin molecule. The eight cysteine

residues (C, shaded) may form four intramolecular disulphide bridges. (Hordijk er
al., 1991c.)
3.1.3. Purifzcation and Structure of the Antagonistic Humoral Factor

Schistosomin
Schistosomin was purified from haemolymph of infected snails and the
complete primary structure was determined (Hordijk et al., 1991b, c).
Schistosomin was shown to consist of 79 amino acids (Figure 14) with a
molecular mass of 8.7 kDa (Hordijk e f al., 1991~).The eight cysteine
residues, which may form four intramolecular disulphide bridges are prob-
ably responsible for the highly folded nature and the rigid three-dimensional
structure of the molecule.
Since the fractions obtained in the schistosomin purification steps were
tested for their biological activity in the CaFl-bioassay, the question
remains of whether one or more types of schistosomin in the haemolymph
are responsible for antagonizing the bioactivity of structurally different
% response
loo 1
80 -
60-
40-
20 -
0 - 1 I I 1
serum 0.0 2.5 5 .O 7.5
protein
pmol schistosomin
Figure 15 Inhibition by schistosomin of the ovulation-inducing activity of
synthetic CDCH in Lymnaea stugnalis. Increasing doses of purified schistosomin
(0-3.5 pmol) were injected simultaneously with 2 pmol of synthetic CDCH.After
30 min the animals were dissected and checked for ovulation and formation of an
egg mass. Haemolymph (serum) protein as well as the buffer from the final
purification step of schistosomin (0 pmol schistosomin) were used as controls.
(Hordijk er al., 1991a.)
female gonadotropic hormones in parasitized snails. To solve this question

HPLC-purified schistosomin was applied in the bioassays developed for
CaFl, CDCH and DBH SyntheticCDCH was injected together with increas-
ing doses of schistosomin in non-infected snails. The response to synthetic
CDCH (2 pmol), ovulation, was blocked for 90% by 3.5 pmol schistosomin
(Figure 15; Hordijk et al., 1991a). In additional experiments it was shown
that purified schistosomin is also capable of irihibiting the biological activity
of CaFl and DBH (Figure 16; Hordijk et: al., 1991d). Apparently the
biological activities of gonadotropic hormones are all blocked by one
and the same schistosomin molecule. An intriguing quehtion now is how
is this performed?
3.1.4. Interaction of Schistosomin with Hormone-Receptor Complexes

In the CaFl-assay indications were obtained that schistosomin antagonizes
CaFl at the level of the hormone-receptor complexes: when albumen
glands were exposed first to schistosomin and then to CaFl, the inhibition
of the effect of CaFl was stronger than in glands exposed to schistosomin
and CaFl at the same time. Furthermore, the response to CaFl in glands pre-
incubated with schistosomin appeared to increase gradually when the
glands were rinsed in Ringer during increasing periods of time before
being exposed to CaFl. This implies that schistosomin is gradually
removed from the receptors (De Jong-Brink et al., 1988b). That the
inhibitory effect of schistosomin on the biological activity of CaFl occurs
at the receptor level was confirmed by using a receptor-binding assay for
CaFl: binding of synthetic CaFl labelled with fluorescein isothiocyanate
(FITC) to a membrane fraction of albumen glands was tested. In the
presence of either haemolymph from infected snails or purified schisto-
Calfluxin DBH CDCH
Figure 16 Effect of purified schistosominon the bioactivity of different gonado-

tropic hormones. The bioactivity of calfluxin, the dorsal body hormone (DBH),
and the caudodorsal cell hormone (CDCH) were tested in the presence of Ringer,
a buffer control from the final purification step of schistosomin, and purified
schistosomin. (Hordijk et al., 1991d.)
HOW SCHISTOSOMES PROFIT FROM STRESS RESPONSES ELICITED IN THEIR HOSTS 21 1
somin the binding of FITC-CaFI to the membrane fraction was inhibited.

However, the conclusion that schistosomin acts at the level of the hormone-
receptor complex does not necessarily imply that both CaFl and schisto-
somin interact with the same receptor (competitive antagonism; Figure 17).
The antagonizing effect of schistosomin might also be exerted by modify-
ing the hormone-specific receptor, resulting in a decrease of the affinity of
the receptor for CaFl (non-competitive antagonism; Figure 17). In view of
the fact that schistosomin antagonizes structurally different gonadotropic
hormones, which are linked to different second messenger pathways, it
seems likely that it exerts these effects by a common mechanism, i.e., after
binding to an own receptor. As a consequence of being bound to the
schistosomin-specific receptor (R2; Figure 17), it might desensitize the
responses to the different hormones (Hordijk et al., 1991b). This corre-
sponds to the two-signal-model proposed by Roszman and Brooks ( 1992)
for the interaction of neurohormones and immune stimuli with their
respective receptors on lymphocytes. It implies that this interaction takes
place at the level of the second messenger cascades linked to the receptor
complexes. This cross-talk can result in either attenuation, as in the case of
schistosomin (physiological antagonism; Figure 17), or in amplification of
Comptetive
anragonism
NOn-
competetive
anragonism
PhySlOlOgICd
Pntapmsm
Rseptor for Receptor for

hormone schrstosomm
Figure 17 Schematic representation of three possibilities for the antagonistic

interaction between a hormone and a schistosomin at the receptor complex level
of a target cell: competitive antagonism, non-competitive antagonism and
physiological antagonism.
the second messengers and subsequent biochemical events. As physio-

logical antagonism does not explain that schistosomin affects binding of
CaFl to its receptor, nor-competitive antagonism might also play a role.
3.2. Central Effects

The possibility that in haemolymph of Parasitized snails humoral factor(s)
occur, which interfere with the activity of neurosecretory cells in the CNS
that synthesize and release the hormones involved in regulation of repro-
duction, growth and metabolism, was studied with both electrophysiological
. and enzyme-cytochemical methods.
3.2.1. Electrophysiological Studies

Based on indications that the neuroendocrine cells controlling reproduction
and growth in Lymnuea show different electrophysiological characteristics
in infected snails compared to those in non-infected controls, the excit-
ability of the CDCs and the LGCs was studied in vitro in the presence of
haemolymph of infected and of non-infected snails.
CDCs display three different states of excitability (the refractory or
inhibited state, the resting state and the afterdischarge) the Occurrence of
which is closely related to the egg-laying cycle (Kits, 1980). Resting-state
CDCs are silent but excitable. Indicative of this state is the occurrence of a
depolarizing afterpotential (DAP) of several seconds following short trains
of electrically evoked action potentials. Prolonged stimulation of resting
state CDCs gives an afterdischarge, whereas inhibited state CDCs are silent
and do not produce an afterdischarge upon electrical stimulation. Resting
state CDCs (in situ in the isolated CNS) in haemolymph of normal, non-
parasitized snails showed the same phenomena on suprathreshold current
pulses as described above. However, after replacing the normal haemo-
lymph by haemolymph from parasitized snails, a strong suppression of the
DAP was observed, and the cells could not generate an afterdischarge
(Hordijk et al., 1992). Similar effects were observed when the CDCs
were bathed in saline with purified schistosomin: at a concentration of
M it reduces the amplitude of the DAP by about 60%.
LGCs in situ in isolated CNS kept in haemolymph of non-parasitized
snails, were silent and responded with a single action potential upon
suprathreshold stimulation. When the haemolymph was replaced by haemo-
lymph from parasitized snails the excitability of the cells increased. The
cells showed a slow-depolarization, a decrease in spiking threshold and a
decrease in the rate of accommodation. In this case, too, purified schisto-
somin mimicked the effect of haemolymph from parasitized snails. The
HOW SCHISTOSOMESPROFITFROM STRESS RESPONSES ELICITEDI N THEIR HOSTS 213
1 schistosornin 2.10-7 M f
Figure 18 Response of a dissociated LGC to 3 min bath application of

2 X lo-’ M schistosomin. Schistosomin induced a slow depolarization of the cell
and a discharge of spiking activity, that outlasted the presence of the peptide in the
bath. In this experiment, schistosomin was applied by means of pressure ejection,
the onset and end of which are indicated by arrows. (Hordijk et al., 1992.)
same phenomena were found using freshly dissociated LGCs in primary

culture (Figure 18; Hordijk et al., 1992). This shows that schistosomin acts
directly upon LGCs and that the response is not mediated by intemeurons
indicating that the LGCs have receptors for schistosomin.
3.2.2. Enzyme-Cytochemical Studies

It was investigated whether the effect of schistosomin on the excitability of
the neurosecretory cells may be mediated by CAMP,that is, by enhancing
the activity of the enzyme adenylate cyclase (AC). In addition, the effect
of schistosomin on AC activity was studied in the endocrine DBs (De
Jong-Brink et al., 1992). In these experiments cerebral ganglia were
excised, incubated in snail Ringer with or without schistosomin for
20 min and processed for the demonstration of AC activity at the ultra-
structural level (Cutler, 1983; De Jong-Brink et al., 1986a). The presence
of lead-imidodiphosphate deposits along the cell membrane indicates AC
activity.
The inhibiting effect of schistosomin on the CDCs appeared not to be
mediated by an increase of CAMP.This is in line with the observation that
cAMP increases the excitability of the CDCs and the release of secretory
material in their neurohaemal area, the cerebral commissure (Buma et af.,
1986; Moed et al., 1989). The enhancement of the excitability of the LGCs
by schistosomin coincides with activation of AC. This result is in keeping
with electrophysiological data showing that an increase in the level of
cAMP increases the excitability of the LGCs. AC was only found along
the highly folded apical cell membrane of these cells, indicating that the
receptors for schistosomin are present in this part of the cell membrane.
The question of whether schistosomin has an effect on DBs could not be
answered since application of schistosomin or of forskolin, an activator of
AC, did not activate the AC-CAMPsystem in DBs. So, if schistosomin has
an effect on DBs, it is not mediated by the AC-CAMPsystem.
The observation that schistosomin exerts both central and peripheral
effects on female reproduction-regulating neuroendocrine mechanisms
justifies the conclusion that it is responsible for the inhibition of egg laying
in L. stagnalis infected with T. ocellata. As schistosomin also has been
demonstrated to enhance AC activity in neurons producing the peptide
alanine-proline-glycine-tryptophan(APGW n e w m in the anterior lobes
of the cerebral ganglia; Figure 12), which are involved in the innervation of
the male copulatory system (Croll et al., 1991), it may alsoplay a role in
the inhibitory effects of parasitosis on male reproductive activity in this
. hermaphrodite snail.
However, the inhibition of the development of the reproductive tract in
juvenile snails, which is already obvious at an early stage of infection,
cannot be ascribed to schistosomin as it was absent from haemolymph of
snails in this stage of infection. Therefore, another mechanism must be
responsible for this effect of parasitosis. The same is true for the increase in
fecundity that was observed in the first week after infection in snails
exposed when sub-adult.
The evidence that schistosomin clearly affects the LGCs suggests that it
is also involved in the effects on growth and metabolism in parasitized
snails. This corresponds to the observation that the start of enhanced body
growth and glycogen depletion in infected snails coincides with the
appearance of schistosomin in the haemolymph. Up until now no assays
are available to investigate whether schistosomin also interferes with the
effects of LGC products on their target tissues.
3.3. Schistosomin: Origin and Induction of its Release by a

Pamito-dorived Factor
3.3.1. Origin of Schistosomin

On the basis of the observation that schistosomin was also only found in
C N S extracts -of both parasitized and non-parasitized snails -and not in
extracts of different parasite stages, it was assumed at first that it originates
from the CNS of the host (Schallig et al., 1991b). This seemed to be
supported by inmunocytochemical studies using a polyclonal antiserum
raised in rabbits against purified schistosomin. Immunopositive material
appeared to & present in the growth controlling LGCs and in the periphery
of the median lip nerve, the neurohaemal area of the LGCs,in the two CCs
of the lateral lobes and in a small number of cells in the pedal ganglia
(Hordijk et al., 1991~).However, the conclusion that schistosomin is a
neuropeptide has appeared to be wrong as shown by the following observa-

tions. With a polyclonal antiserum raised against purified schistosomin in a
guinea pig no immunopositive material was found in the neurosecretory
cells mentioned both at the light and electron microscope level. On the
other hand, small connective tissue cells appeared to contain immuno-
positive secretory material. The cells have thin extensions accompanying
axons to peripheral tissues and organs of the snail which originate from
neurons in the ganglia of the CNS (Figure 19). These cells, which contain
rather big secretory granules, are probably homologous to the “telo-glia”
described for several molluscs by Nicaise (1973). Apparently, certain
eptopes of secretory material in LGCs and CCs had cross-reacted with
the previously used rabbit antiserum. This implies that schistosomin in
CNS extracts originates from cells in the connective tissue sheath and not
from neurosecretory cells. This might explain why schistosomin could also
be extracted from connective tissue at other locations. With the antiserum
raised in guinea pigs connective tissue cells similar to the telo-glial cells
around the CNS, appeared to be the source of schistosomin. However, these
Figure 19 Electron micrographs of a telo-glial cell showing a cell body (cb) and
an elongation (insert) accompanying an axon (a) in the penial complex of L.
srugnulis. The secretory granules (s) show immuno-gold labelling with anti-
schistosomin. n, nucleus.
telo-glial cells are not the only source of schistosomin. Immunocyto-

chemical staining showed that haemocytes from haemolymph of parasi-
tized snails gave a positive reaction with the guinea pig antiserum, whereas
those from haemolymph of non-parasitized snails were almost negative
(Figure 20; Hoek et al., 1994). Schistosomin from haemocytes may also
contribute to schistosomin in extracts of CNS as part 'of the aorta, contain-
ing a large number of haemocytes, was present in the CNS preparations.
The glial cells within the CNS of both non-parasitized and parasitized L.
stagnalis did not show immunostaining with antischistosomin. Although
these cells in the CNS of another freshwater snail, Plunorbarius corneus,
appeared to be capable of phagocytosis, :especially @er damage to the
tissue (Pentreath et al., 1985) and resemble microglia in vertebrates
(Bloom, 1993), they are apparently quite different from haemocytes.
3.3.2. Induction of Schistosomin Release by a Parasite-derived Factor

As the presence of schistosomin in haemolymph of infected snails
coincides with the appearance of differentiating cercariae within the
Figure 20 Light micrographs of monolayers of haemocytes from L. stagnalis

patently infected with T. ocelluta (left) and from a non-infected control (right) both
immuno-stained with antischistosomin. (Hoek et al., 1994.)
daughter sporocysts (see also Figure lo), the stimulation of its production
and release is probably caused by a humoral factor derived from cercariae.
An in vitro bioassay was developed to study the origin and nature of this
parasitic factor. In this bioassay freshly dissected CNS of L. stagnalis were
incubated in media containing acetic acid or methanolic extracts of cer-
cariae and of other developmental stages of the parasite (miracidia and in
vitro cultured mother sporocysts) in order to see whether these extracts
induce the release of schistosomin from CNS in vitro (Schallig et al.,
1992b). HPLC-purified release products of the incubated CNS with chro-
matographic properties of schistosomin were tested for bioactivity in the
CaF1-bioassay. As mentioned, in this assay the stimulating effect of CaFl
on secretory cells of the albumen gland is inhibited in the presence of
schistosomin. Release of schistosomin was only found to be induced with a
methanolic extract of cercariae whereas an acetic acid extract had no
effect. Both acetic acid and methanolic extracts of miracidia and mother
sporocysts did not induce schistosomin release (Figure 21). As only the
methanolic extract of cercariae contains a factor inducing schistosomin
release in vitro, it is likely that the parasite factor has a more or less
Figure 21 Percentages (means and standard deviations) of Ca*+-positivemito-

chondria in albumen glands of non-infected L. stagnalis incubated in Ringer (R), in
Ringer + calfluxin (R+CaFl) or in Ringer + CaFl to which HPLC purified material
was added. A-C: acetic acid extracts of miracidia (A), mother sporocysts (B) or
cercariae (C). D-F methanolic extracts of miracidia (D), mother sporocysts (E) or
cercariae (F). Groups not sharing a common letter differ significantly. (Schallig et
al., 1991c.)
1
hydrophobic character (Schallig et al., 1992b). As schistosomin is already

present in haemolymph of infected snails before patency is reached, viz.,
when differentiating cercariae are present in the daughter sporocyst, this
cercarial factor is either a diffusible molecule or a protonephridial excre-
tion product. In both cases it can pass the wall of the daughter sporocyst.
However, the supposition that eicosanoids, vertebrate type steroids or
ecdysteroids might be good candidates was not confirmed (Schallig, 1991).
This cercaria-derived factor can also induce the release of schistosomin
in viva The response to CaFl was significantly inhibited by haemolymph of
snails injected with a methanolic extract of cercariae compared to haemo-
lymph of control snails injected with only sniU Ringer (De Jong-Brink,
19928). The conclusion that a cercaria-derived factor is responsible for the
induction of schistosomin release and hence for the inhibition of reproduc-
tion, is supported by the observation that reproduction is resumed in
parasitized snails, which had been treated with praziquantel. This drug
only attacks cercariae, leaving the sporocysts intact (Riley and Chappell,
1990).
Addition of a methanolic extract of cercariae to a monolayer of haemo-
cytes from non-parasitized snails also induced schistosomin release into the
medium, as was shown by a dotting immunoassay (DIA) using antischisto-
somin. With a corresponding extract of miracidia no release was observed.
So haemocytes and probably also the telo-glial cells have receptors for the
putative cercaria-derived factor. Furthermore, the amount of schistosomin,
released by haemocytes gradually increased both with increasing numbers
of haemocytes and/or concentrations of cercarial extracts (Hoek et al.,
1994). However, these data need confirmation using Hp2C purification
of the schistosomin peptide.
By using the polymerase chain reaction (PCR),mRNA of schistosomin
has been demonstrated in haemocytes from both parasitized and non-
parasitized snails. As no obvious difference was found in the mRNA
quantities, induction of schistosomin synthesis/release seems to be
regulated at the translation level (Hoek et al., 1994).
The results obtained from schistosomin strongly suggest that it repre-
sents a cytokine(-like) molecule in Lymnaea, because it meets the criteria
as summarized by Beck and Habicht (1991a). It is a polypeptide mediator
produced by cells of the internal defence system (haemocytes) and by non-
immune (?) telo-glial cells and it is released by these cells in response to a
special stimulus (cercarial factor) in a dose-dependent way. With regard to
the criterion that it should have an effect on cells of the immune system,
results have been obtained showing that schistosomin suppresses haemo-
cyte activity (Hoeket al., 1994). The conclusion that schistosomin might
represent a cytokine-like molecule is also in line with the fact that it
interferes with different neuroendocrine systems regulating processes
HOW SCHISTOSOMES PROFITFROM STRESS RESPONSES ELICITEDIN THEIR HOSTS 219
such as reproduction and growth and that it exerts its effect at two
levels: the productionhelease of hormones and their effect upon target
cells. However, schistosomin shows no sequence homology with any of
the vertebrate types of cytokines identified so far. It has been demonstrated
that a factor acting similarly to schistosomin occurs in haemolymph of
other schistosome-snail combinations, e.g., S. mansoni-B. glabrata (De
Jong-Brink et al., 1991). As haemolymph from L. stugnalis infected with T .
ocellatu did not affect the CaFl response in albumen gland cells of B .
glubratu and similarly the response to CaFl in L. stagnalis was not
inhibited by haemolymph from B . glabrata infected with S . mansoni, it
seems interesting to study the degree of sequence homology between the
factors in haemolymph of different species of parasitized snails.
4. PARASITES: LONG-TERM STRESSORS?
4.1. The Stress-concept in Mammals
The actions exerted by schistosomin on the NEB in snails are comparable

to those exerted by inflammatory cytokines (11-1, 11-6, TNF, Stadnyk and
Gauldie, 1992) on the hypothalamic-pituitary-adrenal (HPA) axis in
mammals (Berkenbosch et al., 1987; Sapolsky et al., 1987; Stemberg et
ul., 1989; Rothwell, 1991). The activation of the HPA axis is one of the
most important and extensively studied effects induced by stress.
The physiological effects evoked by schistosome parasites in their snail
host strongly resemble those caused by other kind of insults which disrupt
the physiological balance and evoke aspecific stress responses in mammals.
These stress responses can be considered as an adaptation to re-establish
the balance (Sapolsky, 1992). At first, processes are suppressed which are
initially not important or even disadvantageous for the infected organism,
such as inflammation, pain perception (analgesia), immune function, food
intake and digestion, growth and reproduction. Others are enhanced or
show remarkable changes, e.g., metabolism (stored energy becomes avail-
able), the cardiovascular activity (more of the mobilized glucose and
oxygen is delivered to the tissues, the intestinal tract excluded) and the
kidney water resorption (the blood volume is increased). It has become
evident that all these adaptational stress responses in mammals are brought
about via the hypothalamic-pituitary (HP)axis and the autonomic nervous
system. Stressors, that is, anything that disrupts the physiological balance
(Sapolsky, 1992), are perceived by the brain, either via sensory nervous
pathways and/or by the action of cytokines released from activated
macrophages. This leads to stimulation of some neuroendocrine systems in

the H p axis and of the sympathetic part of the nervous system and to
inhibition of other neuroendocrine systems of the HP axis and of the
parasympathetic part of the nervous system resulting in changed balances
of hormones and other regulatory substances and hence in physiological
adaptations to stress. The stimulated sympathetic nervous system releases
noradrenaline from its endings, except for the projection extending into the
adrenal medulla, where adrenaline is released which in turn stimulates the
release of adrenaline from the medullar cells of the adrenals (Navarra et al.,
1991).
In the context of this review, the effects of stress on the immune
,system and reproduction and growth in mammals will be summarized
briefly.
4.1.1.Stress and the Immune System

The suppressive effect of stress on the immune system can partly be
ascribed to the increased level of glucocorticoids. Stress stimulates the
release of corticotropin releasing factor (CRF) from the hypothalamus.
CRF induces the release of adrenocorticotropic hormone (ACTH) from
the pituitary, that subsequently stimulates the adrenal cortex to release
glucocorticoids. These steroids appear to inhibit cells of the immune
system to release cytokines and to lower the sensitivity of target cells to
them, which finally results in an inhibition of proliferative responses of the
immune system cells (Bateman et al., 1989). However, this is not the only
link between stress and the immune system, which might explain the
immunosuppressive effects of stress. Many other hormones are known to
influence immune responses. The immunosuppressiverole of opioids, which
aie derived from the common precursor molecule pro-opiomelanocortin
(POMC) and secreted by the pituitary during stress, is illustrative in this
respect. The presence of opioid receptors on various cells of the immune
system (cf. Carr, 1992) suggests that the immunosuppressive effects of
opioids upon these cells are direct. These effects of POMC-derived pep-
tides might counteract some immunosuppressive effects of glucocorticoids,
and prevent any overshooting of the glucocorticoid-dependent effect on
immune cells, as shown for natural killer cell activity (Gatti et al., 1993).
The situation becomes even more complex as POMC-derived peptides
also originate from lymphocytes. CRF stimulates macrophages to produce
Il-1, whereas 11-1 elicits POMC peptide production by B lymphocytes
(Kavelaars eral., 1989);
Activation of the sympathetic nervous system also has an immuno-
modulatory effect. Neuropeptide Y (NPY),a neurotransmitter present in
HOW SCHISTOSOMES PROFIT FROM STRESS RESPONSES ELICITED IN THEIR HOSTS 22 1
peripheral sympathetic nerves, is supposed to play an important immuno-

regulatory effect during stress. In vitro experiments have confirmed this
immunoregulatory role of NPY on natural killer activity of normal human
lymphocytes (Nair et al., 1993).
Yet another communication link between the peripheral part of the CNS
and the immune system is the innervation of lymphoid organs, which
comprises abundant noradrenergic and peptidergic elements (Felten et
al., 1992). Stimulation of the noradrenergic nerve endings innervating
immune tissues, as is the case during stress, results in immunosuppres-
sion, whereas destruction of these projections seems to enhance immune
activity (Sapolsky, 1992). Also in this case the presence of appropriate
adrenergic and peptidergic receptors on cells of the immune system might
indicate the important role of these nerve fibres in the communication
between the two regulatory systems.
4.1.2. Stress and Reproduction and Growth

Inhibition during stress of several hormones involved in regulation of
reproduction and associated behaviours can be explained by the fact that
stress affects the hypothalamic-pituitary-gonad (HPG) axis. Secretion of
gonadotropin-releasing hormone (GnRH) in the hypothalamus is inhibited,
which results in a decreased release of luteinizing hormone (LH) and
follicle-stimulating hormone (FSH) from the pituitary and subsequently
of sex steroids from the gonad. These sex steroids also have an effect on
the immune response. Androgens tend to suppress both humoral and
cell-mediated responses, whereas oestrogens enhance humoral responses.
An activation of the immune response in males by the lowered concentra-
tion of androgens is, however, counteracted by the increased level of
glucocorticosteroids. Furthermore, as the capacity of the receptors for
glucocorticoids is higher than that of receptors for sex steroids, glucocorti-
coids may have a stronger effect on the immune system with suppression as
a result (Alexander and Stimpson, 1988).
Data on the effects of stress on growth are rather confusing, because the
picture is not the same for all mammals investigated. The release of growth
hormone (GH) from the pituitary is either promptly inhibited by stressors,
as has been described for rodents, or initially enhanced, as is the situation
in humans. In the latter the GH release is inhibited by persistent stressors.
The secretion of somatomedins by the liver reflects that of GH release. The
effects on GH release are established via the release of growth hormone
releasing hormone (GHRH) and its antagonist somatostatin from the
hypothalamus.
4.2. T. ocellata: a Stressor for L. stagnalis?
4.2.1. Role of IDS in NEgResponse

It is striking that in mammals when they are stressed by an infectious
challenge, suppression of the immune system by glucocorticoids is pre-
ceded by an activation of macrophages, which Sapolsky (1992) indicated
as “the system asks for being suppressed”. In snails, which become
infected with T. ocellata there was an initial activation of haemocyte
activity, followed by a suppression. Involvement of haemocytes in stress
responses in molluscs has also been emphasized by Stefan0 et al. (1990)
and Ottaviani et al. (1992, 1993a).
Also the effects on reproduction and growth caused by T. ocellata in its
snail host and on the neuroendocrine mechanisms underlying it show great
similarities with the effects of stress in mammals and support the idea that
the parasite T. ocellata can be considered as a stressor for its host evoking
aspecific stress responses. This would imply that similar stress responses
can be evoked in Lyrnnaea by other stressors/stress situations and that
schistosomin, which plays a crucial role in establishing the effects on
reproduction and growth in parasitized snails, is also released in non-
parasitized snails when they are confronted with other adverse conditions
which are known to stop reproduction. This supposition was studied by
taking haemolymph from three groups of snails, which had stopped egg
laying because of different conditions: (1) starved for 12 days, (2) kept for
12 days at low temperature and (3) kept for 5 days in water dirtied by
crushed rotting lettuce. Testing of the haemolymph samples from these
snails in the Cam-bioassay for the presence of schistosomin revealed that
they do not contain schistosomin in such amounts that it inhibits the
hormone response significantly (Figure 22; De Jong-Brink et al., 1992).
Therefore, other mechanisms must be involved in the inhibition of repro-
duction in snails under these “long-term” adverse conditions.
For this reason, it was also investigated whether schistosomin is involved
in ,establishing “short-term” inhibitory effects on reproduction, e.g.,
reflected by the sometimes observed phenomenon that the time needed
for the production of an egg mass is longer in snails which have been
disturbed while producing an egg mass.
Experiments have been performed in which snails were disturbed for a
short time after they had received a stimulus for ovulation/egg-mass
deposition, i.e., after their neurosecretory CDCs had started their firing
activity (Kits, 1980). The latter was evoked by transferring snails, which
had been kept in dirtied water for 4-5 days, to clean oxygenated water (the
clean water stimulus, CWS;Ter Maat et al., 1983). Groups of snails were
disturbed either before or after they had received the CWS by loosening
100
90
80
70
60
30
20
10
0
A B C D E F
Figure 22 Means and standard deviations of the percentages of Ca-positive
mitochondria in albumen glands of L. stagnalis incubated in the following
media: A, Ringer (R); B, R with the neuropeptide calfluxin (R + CaFl); C, R +
CaFl + schistosomin; D, haemolymph from snails starved for 12 days + CaFl; E,
haemolymph from snails starved for 12 days + CaFl; E, haemolymph from snails
kept for 12 days at 4°C + CaFl; F, haemolymph from snails kept for 45 days in
dirtied water + CaFl. Each mean is based on counts in five glands, 100 mito-
chondria per gland. Groups not sharing a common letter differ significantly. (De
Jong-Brink et al., 1992.)
them from the wall of the jar and placing the jar with the snail on a slowly
moving shaker for 2 min in order to prevent the snail from adhering again
to the wall. All the disturbed groups of snails showed a significant increase
of the time (ca 30-40 min) between the CWS and the moment of ovulation
egg-mass deposition compared to the non-disturbed controls, irrespective of
their being disturbed before or after the CWS. This indicates that this delay
of egg-mass deposition is induced independently of the CWS, that, is
activation of CDC activity.
The question of whether this delay was caused by the release of schis-
tosomin into the haemolymph of disturbed snails was investigated using
three bioassays, (1) the CaFl bioassay, (2) a dotting immuno assay (DIA)
using a polyclonal antiserum against schistosomin and (3) an assay in
which the excitability of isolated LGCs (mean number of action potentials
per 10 s) was quantified (De Jong-Brink et al., 1994). As mentioned,
schistosomin causes an increase of LGC excitability. Haemolymph of
disturbed snails with delayed ovulation or oviposition, showed similar
effects as purified schistosomin, whereas this was not the case with
haemolymph from non-disturbed controls. As the release of schistosomin
was also obvious in snails, which had been disturbed while staying in
dirtied water, the question remains whether such a disturbance might
also induce schistosomin release in snails which had been kept under
75 C C
11 I
b
1
0
a
1
ap
0 A - - - -
B C D E
Figure 23 Percentages (means and standard deviations) of Ca2+ (Ca-)positive
mitochondria in albumen glands of non-infected L. srugnalis incubated in Ringer
(A), in Ringer with calfluxin (CaF1, B) or with CaFl in haemolymph from snails
30 min after an ovulation-inducing clean water stimulus (CWS) (C), 30 min after
CWS, 20 min after disturbance (D), 90 min after C W S (E),90 min after CWS,
30 min after disturbance (F).Groups not sharing a common letter on top of the bars
differ significantly. (De Jong-Brink er al., 1994.)
normal conditions. Also the haemolymph of these snails gave a significant

decrease of the hormone response in the CaF‘Lbioassay compared to that of
non-disturbed controls indicating the presence of schistosomin. However,
the hormone response was significantly less inhibited than with haemo-
lymph from snails which had been disturbed in dirtied water (Figure 24).
Also in snails (kept under normal conditions), in which ovulation was
induced by injecting them with an extract of the COMs, the neurohaemal
area of the CDCs, disturbance either before ovulation or during packaging
of ‘eggs into an egg mass resulted in a delay of ca 30 min, which coincided
with presence of schistosomin in the haemolymph. On the other hand,
snails injected with Ringer only also showed release of schistosomin into
the ,haemolymph. Apparently, the injection per se already evokes a weak
stress response. The amount of schistosomin is, however, subthreshold it
does not delay ovulation/egg laying, which becomes apparent when an
extract of COMs (CDCH) is co-injected.
These data clearly demonstrate that in snails disturbed (stressed) before
or after the onset of an egg-laying episode, schistosomin is released into the
haemolymph and causes retardation of ovulation/egg-mass deposition.
Unfavourable conditions such as “dirtied water” and “being injected”
may induce the release of schistosomin, but not in such high amounts
that it delays ovulation and egg laying. However, if these snails are also
disturbed, the concentration of schistosomin surpasses a threshold, and
HOW SCHISTOSOMES PROFITFROM STRESSRESPONSES ELICITEDIN THEIR HOSTS 225
1
T
R R+ NC NC DW DW
CaFl +D +D
Figure 24 Percentages (means and standard deviations) of Ca2+ positive (Ca

pos) mitochondria in albumen glands of non-infected L. stagnalis incubated in
Ringer (R), in Ringer with the hormone calfluxin (CaFl), or in haemolymph - with
CaFl - from snails ( 1 ) kept under normal conditions (NC), (2) NC + disturbed (NC
+ D), (3) kept in dirtied water (DW), (4) DW + disturbed (DW + D). Groups not
sharing a common letter on top of the bars differ significantly. (De Jong-Brink et
al., 1994.)
becomes high enough to cause a delay of ovulation/egg-mass deposition.

The fact that this delay was ca 30-40 min in all snails, which had been
disturbed in the same way, might indicate that in this period either the
number of receptors for schistosomin or their sensitivity has decreased and/
or that inactivation of the schistosomin molecule itself occurs.
Apparently, T. ocellata makes use of this short-term stress mechanism in
its snail host and can be considered as a stressor. By forming cercariae
incessantly, the parasite continuously induces haemocytes and possibly
also connective tissue cells to release schistosomin so that reproduction
of the host remains inhibited and energy is available for its own develop-
ment. Under these long-term or chronic stress conditions schistosomin is
probably also responsible for the increased - be it abnormal - growth of
the host as it appeared to stimulate the LGCs in the CNS, which are
involved in regulation of growth.
4.2.2. Role of NES in IDS Response

As mentioned, opioid peptides, secreted during stress, are important modi-
fiers of the IS. Also in molluscs and other invertebrates, opioids have
been found to influence haemocyte activity (Stefan0 et ul., 1989a, b, 1991a,
b, 1993). The observation that POMC-derived peptides were detected in
haemolymph of S. mnsoni-infected B. glubrutu (a-melanocyte stimulatory
hormone, a-MSH and to a lesser extent ACTH and P-endorphin), whereas

they could not be detected in haemolymph of control animals, suggests a
role in immunosuppression (Duvaux-Miret et al., 1992). The question is
whether these POMC-derived peptides originate from the parasite -which
possesses a POMC-related gene - or from the host. The first possibility is
supported by the observation that co-incubation of aduit worms with human
polymorphonuclear leucocytes or with B. glabrata haemocytes led to the
appearance of a-MSH in the medium, probably resulting from a conversion
of parasite ACTH. However, it is also possible that they are released by the
defence cells themselves. Haemocytes in the closely related mail Planorbis
corneus appear to contain material immunoreactive with antisera to
POMC-derived peptides (Ottaviani et al., 1990). In a later study Duvaux-
Miret et al. (1993) reported a metenkephalin-like peptide to be present in
adult worms and in their incubation medium. It seems interesting to trace
the origin of this peptide in the parasite. The possibility that it originates
from the nervous system seems unlikely.
A parasitic origin of these peptides in the snail host seems doubtful as
immunocytochemical staining with antisera raised against POMC-derived
peptides only revealed anti-metenkephalin positive material in undifferen-
tiated cells in sporocysts of T. ocellata and S . mnsoni. As neither the
tegumental cells nor glandular cells showed positive staining, it seems
rather unlikely that this metenkephalin-like material will be secreted into
the haemolymph of the host (Solis Soto and De Jong-Brink, 1994). More-
over, the question of whether schistosomes induce the nervous system and/
or haemocytes of their hosts to release POMC-derived peptides is still open.
Antisera raised against vertebrate opioid peptides caused positive stain-
ing in neurosecretory cells in the CNS of Lymnaea (Boer et al., 1987).
Therefore, the possibility was investigated that E/S products of T. ocellata
induce the snail CNS to release neuropeptides, e.g., opioid peptides, into
the haemolymph, which subsequently might be responsible for plasma-
associated immunosuppression in the snail host. A medium, in which CNS
had been incubated with E/S products released by 72 or 96 h cultured T.
ocellata appeared to suppress phagocytic activity of haemocytes compared
to a control medium, in which CNS had been incubated without E/S
products (Figure 25) (De Jong-Brink et al., 1991; Amen and De Jong-
Brink, 1992). It is, however, still questionable whether neuropeptides are
responsible for this effect, as in later, unpublished, experiments similar
results were obtained with connective tissue samples. Also in this stage of
infection cytokine-like molecules derived from glial cells within the CNS,
from connective tissue cells or from haemocytes - these cells are
transferred with the CNS to the incubation media - might be
involved. This would imply that the early effects evoked by miracidid
mother sporocysts are mediated in a similar way to the later effects elicited
260 -
230 -
-
i t +
200
170 -
140 -
110 -
80 - I I I I I I I
Figure 25 Means and 95% confidence intervals of the number of haemocytes

(out of 500) from non-infected snails (n = 20) phagocytosing zymosan particles
after incubation (30 min) in the following media: 1. plasma of non-infected
snails, 2. minimal salt medium (MSM), 3. MSM in which CNS had been
incubated (4-6 h), 4. MSM in which mother sporocysts had been cultured
(4 days), 5 . Medium 4, in which subsequently CNS had been incubated
(4-6 hours). (De Jong-Brink et al., 1991.)
by differentiating cercariae, although the mediating host factor is not

identical to schistosomin.
4.3. Schistosomes: Stressors for Their Vertebrate Hosts?
Based on the results indicating that T. oceffutuis perceived by the snail

host as a stressor, it might be interesting to investigate whether the
physiological effects brought about by schistosome infections in the defi-
nitive host may also, partly, be explained as resulting from stress elicited
by the parasite. This can, however, only be examined in experimental
schistosomiasis models as in naturally infected animalshumans confound-
ing factors such as co-infection with other pathogens, malnutrition and the
infection-associated syndrome cachexia (Tracey and Cerami, 1992) cannot
be excluded.
Moreover, most studies focus on the effect of schistosomes on the IS of
the host (e.g., Stadecker, 1992). Data concerning the effects on the NES
and on links between the two regulatory systems are rare and fragmentary.
Augmentation of the endogenous opiate system has been observed by

Kavaliers et al. (1984) in S . munsoni infected hamsters. This observation
is in line with the idea tha6parasites affect the HP axis. In these studies the
elevated opioid levels were related to changes in locomotory activity of
the host (Kavaliers and Podesta, 1988; Kavaliers and Colwell, 1992). The
elevated levels of opioids, as observed by Kavaliers et al. (1984) might,
according to Isseroff et al. (1986) also be responsible for the lowered levels
of androgens in S . munsoni infected mice, resulting in sexual dysfunction
in chronic schistosomiosis. Data obtained in later experiments suggested
that P-endorphin was responsible for these effects via the HFG axis
(Isseneroff et al., 1989). The lowered levels of androgens are supposed
to be responsible for the decreased levels of mRNA of major urinary
proteins in the liver of schistosome-infected mice (Isseroff et al., 1986).
Thus there are indications, although fragmentary, that schistosomes
affect their definitive host by eliciting stress, responses. The stress
responses might be triggered by inflammatory reactions to schistosomula
and eggs comprising activation of macrophages. In the definitive host the
effects of schistosomes on the H p axis become mainly manifest as suppres-
sion of the IS. The lowered levels of sex steroids, which might lead to a
relief of immunosuppression, is counteracted by increased levels of gluco-
corticoids and opioid peptides. As a survival strategy of the parasite this is
of the utmost importance in view of the advanced development of the IS in
the definitive host. Effects on reproduction and growth via the NES are less
conspicuous except when severe pathology is involved.
4.4. Non-schistosome Parasite-Host Combinations
4.4.1. Effects of Parasite-derived Factors on NES Host

During the last decade many reports have been published on non-
schistosome parasite4ost combinations demonstrating that the physio-
logical effects evoked by parasites in their host are established by the
parasites interfering with the NES of the host (for review see Beckage,
1993b). A few examples will be placed in the context of this review.
(a) Parasite factors mimicking host regulatory substances. The interac-
tion with the NES of the host can be established by the parasites secreting a
factor, which strangly resembles and/or mimics the action of a regulatory
substance or hormone in the host, thereby changing the hormonal balance
in their host.
A very interesting and extensively studied example of such a hormone-
like factor is the factor released by plerocercoid stages of the tape worm,
Spirometra mansonoides (Phares, 1992), which is a homologue of human
growth hormone (hGH). This plerocercoid growth factor (PGF) is probably

an integral membrane protein as a detergent such as Triton X-100 is
necessary for efficient extraction (Phares, 1988). Although PGF has a
high degree of homology with hGH, it does not induce the same effects.
Notwithstanding the reduction of the host’s endogenous production of
growth hormone (GH) by feedback mechanisms, the parasite induces
acceleration of growth in its hosts. The selectivity of action of PGF results
in multiple physiological changes in the host that benefit the parasites. It
specifically binds to highly selective hGH receptors on human lymphocytes
(Phares and Watts, 1988). The transient suppresion of some aspects of the
immune response by PGF as observed in rats may be indicative of its role
in the extremely low host resistance to plerocercoid infections (Phares et
al., 1990; Phares, 1992). How this factor is detached from the membrane in
vivo is not clear. Proteolytic enzymes are supposedly involved.
Another example of a hormone-like factor secreted into the host is the
material in the (sub)tegumental cells of the adult trematode Echinostoma
liei, which shows immunopositive staining with anti-vasoactive intestinal
peptide (VIP Thorndyke and Whitfield, 1987; Thorndyke, 1990). It is
suggested that this parasite-derived VIP is involved in relaxation of gut
smooth muscles and in stimulation of fluid and electrolyte secretion in the
gut lumen. In case these parasites damage the intestinal wall, this VIP-like
material may also stimulate macrophages (Segura et al., 1992).
The release of diffusible ecdysteroids from parasitoids (insects) into their
insect host, lepidopteran larvae, has been demonstrated in in vitro experi-
ments with the hymenopteran Ascogaster reticulatus Wanabe (Brown et al.,
1993). In vivo, this release of ecdysteroids into the insect host supposedly
disturbs the endocrine status of the host. Such a release from parasitoids has
also been suggested for JH compounds but there is no evidence that it does
occur. The JH homologues produced in wasp parasites may even differ from
those of the lepidopteran host (Lawrence and Lanzrein, 1993).
(b) Parasite-factors interfering with production and action of host
regulatory substances. The effects exerted by parasites on their host’s
NES may also be mediated by a parasite-derived factor. Parasitic castra-
tion of crabs parasitized by rhizocephalans (both parasite and host are
crustaceans) can (partly) be explained by the direct action on the NES of
the host exerted by a toxic secretionproduct, a pmteinacems component of
about 25 kDa, released from the roots (rhizoids) of these parasites into the
crab host. This, as yet unidentified protein, is probably released by the
epidermal cells lining the roots by a holocrine secretion process (Rubiliani,
1983; Rubiliani and Payen, 1988).
Products secreted by parasitoids are involved in establishing the effects
on development and reproduction of their host larvae via the NES,that is, by
affecting enzymes either involved in hormone synthesis or in their
degradation. Teratocytes, single cells originating from the serosa (embryo-

nic envelope) of the hymenopteran parasitoid Microplitis croceipes, inhibit
the JH-degrading enzyme JH-esterase in the haemolymph-of its host,
Heliothis virescens larvae (Lawrence, 1990). These effects on JH-esterase
activity are mediated by a host-derived peptide (Hayakawa, 1992). As a
result the JH titre remains high and the development of the host is arrested
(Dahlman, 1990). Parasitoids may also affect the activity of the enzyme
ecdysone 20-monooxygenase in their host, which converts the precursor
3-dehydroecdysone into ecdysone. It is, however, not known how para-
sitoids exert these effects on this enzyme. So, in parasitized insects
hormonal changes may arise from parasitic effects on the activity of
enzymes derived from the fat body, which play a role in either synthesis
or degradation of hormones. Depression of host ecdysteroid titres below the
threshold needed for a moult in parasitized larvae may also be caused by a
lack of release of prothoracicotropic hormone from neurosecretory cells in
the brain-retrocerebral complex (Beckage et al., 1994; Zitnan et al., 1994).
This is based on the observation that multiple neurosecretory material is
accumulated in tobacco hornworm larvae parasitized by the wasp Cotesia
congregata. A parasite-derived factor might induce these effects.
The situation in parasitoid-insect host larvae is even more complicated
as venom and virus-laden calyx fluid (polydnaviruses, PDV) are also
involved in these parasite-host interactions. These “third party” c o m p
nents are injected by female wasps during oviposition. The PDV are
supposed to encode for the parasitism specific “early” proteins in haemo-
lymph of parasitized larvae (Beckage, 1993a). Venom and calyx fluid play
an important role in the inhibition of pupation in a parasitized army worn.
Wani et al. (1990) have shown that inhibition of metamorphosis, i.e.,
prolongation of the larval life, of the host Pseudoletia sepai-ata by terato-
cytes of the parasitoid Apanteles kariyai was only complete when venom
and calyx fluid had been injected previously. Retardation of male repro-
duction, i.e., of testis development and spermatogenesis, as occurs in
parasitized larvae could also be seen in non-parasitized male hosts after
being injected with calyx fluid (PDV particles) and venom (Yagi and
Tanaka, 1992). In this case teratocytes are apparently not involved.
Dushay and Beckage (1993) suggest that the mechanisms by which PDV
act to disrupt development (moulting) in the host larvae may involve the
brain-neurosecretory axis or factors affecting regulation of the JH titre-
Venom components have been shown to promote uncoating of PDV
particles and their penetration into host cells and to potentiate the PDV
effects (Strand and Dover, 1991; Tanaka and Vinson, 1991). The role of
parasitism specific, probably host-derived, “late” peptides or proteins in
haemolymph of insect hosts in affecting the host’s NES has not been
studied (Beckage, 1993a).
Inhibition of female reproduction also occurs in another parasite-insect

combination: metacestodes of Hymenolepis diminuta in the insect Tenebrio
molitor. This effect, however, is not reflected by lowered titres of JH or by
increased JH-esterase activity. It is probably exerted at the receptor level in
the target organs (Hurd, 1993).
These examples show that in many cases parasites are able to disturb the
NES in their host, which may result in physiological changes in the host,
especially with respect to reproduction, growth and development. How-
ever, the mechanisms used by parasites to influence the host’s NES are far
from uniform.
4.4.2.Effects on NES via IDSIIS

(a) In insect hosts. The role of the IS or IDS in establishing the effects
on the NES of the host has, to our knowledge, not been studied. Such an
involvement of the IDS/IS is unlikely in case the parasite influences the
host’s NES by secreting products, which are identical to or mimic a
regulatory component of the host. In other cases, as in insect larvae
hosting parasitoids, the products secreted by teratocytes and proteins
encoded by PDV affect both the hormonal balance, within the PTTH-
ecdysone-JH system, and the IDS in their host. Teratocyte-derived pro-
ducts play a role in immunoevasion. Products of young teratocytes of the
gregarious parasitoid Cotesia glomerata protect it from encapsulation by
IDS cells of the host by inhibiting the activity of the enzyme phenoloxidase
.(Kitano et al., 1990; Tanaka and Wago, 1990). This enzyme plays a major
role in non-self recognition by the insect host (Soderhall and A s p h , 1993).
PDV blocking effects on the immune system of the host larvae have been
reviewed by Stoltz (1993).
As interference by parasites with the regulatory systems in their insect
host has mainly been studied with respect either to the NES or to the IDS,
knowledge concerning possible (effects on) interactions between the two
systems is virtually lacking. Although rather speculative, it can be supposed
that such a relationship between the two systems does exist. Also haemo-
cytes of insects have appeared to release signalling, cytokine-like factors,
which might play a role in this interaction. These are the encapsulation
promoting factor (Ratner and Vinson, 1983), the phagocytosis stimulating
mediator (Mohrig and Shittek, 1979) and hemolin, an immunoglobulin-like
molecule (Sun et al., 1990). Other candidates are the parasitism-specific
“early” and “late” proteins. These are supposed to be unique to parasitized
insect larvae and not inducible by other forms of “physiological stress”,
pathogenic infection or physical trauma and injury (Beckage, 1993). It
seems, however, questionable that the host larvae would respond to para-
sites by synthesizing proteins, the “late” proteins, which do not occur and
have no function under other physiological conditions. It is also unknown

whether in insects hormones/factors from the NES have an effect on the
IDS. Vinson (1993) does not expect the hormones of the PTTH-ecdysone
JH system to influence the IDS. This supposition is based on the discon-
tinuous activity of this hormonal system involved as it is in regulating
discontinuous processes: growth, development and reproduction.
(b) In vertebrate hosts. In the following examples a role of the IS in
establishing the effects of parasites on the NES of their vertebrate host has
not been shown but seems plausible.
Extensive cutaneous myiasis in sheep arising from the activity of larvae
of the insect Lucilia cuprina coincides with signifant changes in the plasma
Foncentrations of (immunoreactive) p-endorphin, ACTH and cortisol
(Schutt et al., 1988). These changes are comparable to the response to
acute-restraint stress except for those in the levels of p-endorphin, which
are reduced in these infected sheep and elevated in acute-restraint stress.
Schutt et al. (1988) explain this difference either by an alteration of
precursor processing in the pituitary or by the selective release of
ACTH. They have not studied the role of inflammatory cytokines. The
elevated levels of catecholamine in turkey poults in the acute stage of
infection with the coccidium Eimeria adenoeides are also indicative for a
classical stress-elicitedresponse: the levels of adrenaline and noradrenaline
were elevated compared to the non-infected controls (Augustine and
Denbow, 1991). The high levels of adrenaline may contribute to the
changes in cardiovascular activity in infected poults. None of the other
hormones of the HP axis were measured.
Since the early 1950s, it has been known that African trypanosomes cause
reproductive disorders in their host resulting from hormonal imbalance.
Hypothyroidism and hypogonadism are very obvious. However, not all
endocrine disorders can be explained as being the result of affecting the
axis. Unmedicated sleeping sickness has appeared to be associated with
adrenal insufficiency and with elevated plasma levels of the cytokines
TNFa and 11-6. The impairment of the adrenocortical function may be
caused by these elevated cytokine concentrations (Reincke et al., 1994).
Also the inhibited gonadal function in African trypanosomiasis is indica-
tive of a functional defect of the HP axis, caused both at the central and
peripheral level of the NES. At the acute phase of infection with Trypano-
soma brucei male rats show a significant decrease of serum LH and
testosterone (Soudan et al., 1992). This dysfunction of the testosterone-
producing Leydig cells in the interstitium of the testis is due to a lack of
responsiveness to human chorionic gonadotropin (hCG, this binds to the
same receptors as LH), which is probably related to the activation of the
macrophages in the interstitium of the testis resulting in an increase of
testicular 11-1 and prostaglandins. Therefore, these effects on the hormonal
balance are supposed to be a local stress effect, mediated by the IS at the

target level. Feedback mechanisms may be responsible for changes at the
level of the corresponding part of the HP axis.
Calves infected with Sarcocystis cruzi, a Coccidia-like protozoan, show
altered growth and metabolism, which may arise in part from infection-
mediated effects on the regulation of GH, somatomedin C and its binding
protein (Elsasser et al., 1988).
The depressed thyrotroph and thyroid gland function during acute,
severe malaria in adult humans (Davis et al, 1990) might result from
stress acting at the pituitary-thyroid axis. The same holds true for the
changed pituitary-adrenal function (Brooks et al., 1969).
Recently, there is mounting evidence that nematode-induced inflamma-
tion in the rat intestine causes alteration of the enteric nerves in the
myenteric plexus as, for example, changes in neurotransmitter release
(Collins et al., 1992) and remodelling of nerves (Stead, 1992). These data
suggest a dynamic interplay between IS and peripheral NES during inflam-
matory episodes. Nothing is known about the consequences of this inter-
play for the central part of the nervous system. It is, however, rather
difficult to define the IS as cytokines secreted by fibroblasts are also
involved in intestinal inflammatory reactions (Pang et al., 1994).
These examples of parasitized vertebrates show that hormonal
imbalances, some of them similar, others dissimilar to classical stress
responses, underlie physiological changes evoked by parasites in their
host. These changes at the level of the central and peripheral part of the
NES of the host are probably mediated by cytokines released during the
inflammatory stages of parasitic infections. Amplifications of the parisitic
signals by cytokines from especially macrophages might explain that even
a very small number of parasites can induce such responses of the NES.
Several explanations can be given for the fact that deviations of the
general stress-elicited responses have been observed in parasitized animals,
both at the level of the IS and the NES. These deviations may be related to
the nutritional status of the host and related effects on metabolism and to
the pathology of infection (cf., Elsasser et af., 1988). Intraspecific diversity
of the host in susceptibility for parasitic infection as well as in the response
to stress (Nozaki and Dvorak, 1993) may play a role. A failure to release
normal levels of corticotropin-releasing factor in response to stress, that
appeared to be correlated with susceptibility to experimentally induced
autoimmunity (Sternberg et al., 1989), might also alter the stress responses
to parasitic infections. Changes in plasma components acting as mediators
of immune responses - acute phase proteins, cytokines, antibodies during
parasitic infections - may also interfere with or counteract stress-related
activation of the HP axis and the resulting physiological effects (Gaillard et
al., 1990; Reincke et af., 1994). In African trypanosomiasis TNF or
cachexin has cachexia-promoting properties (Beutler and Cerami, 1988).

Another example of plasma proteins which interact with physiological
phenomena are the antisecretory factors, which are probably synthesized
in the central nervous system and are, as part of an acute phase response,
released from the adenohypophysis into the blood. They counteract the
pathological intestinal secretion of water and ions during parasitic
infections, for example S. mansoni infections (Nilsson et al., 1992).
These data indicate that parasitic infections have complex effects on
their hosts, which makes it rather difficult to isolate the effects of aspecific
stress responses in parasitized animals.
5. SUMMARY AND CONCLUSIONS
Results obtained with the model Trichobilharzia ocellata-Lymnaea stag-

nalis have confirmed the hypothesis that the physiological effects evoked
by schistosomes in their snail host - castration and giant growth - are
brought about by them interfering with the neuroendocrine systems (NES)
regulating the physiological processes concerned. As soon as differentiat-
ing cercariae are present in the daughter sporocysts a factor can be detected
in the haemolymph of the snail host, called schistosomin, which acts both
at the central and the peripheral parts of the NES involved in regulation of
reproduction and growth. Schistosomin appears to be a host-derived factor,
which is probably released by cells of the internal defence system, the
haemocytes, and by connective tissue cells, the telo-glial cells. It meets the
criteria of having a cytokine-like function although its molecular structure
does not show sequence homology with any of the vertebrate-type cyto-
kines identified to date. Its cytokine nature explains why schistosomin can
interfere with different neuroendocrine regulatory systems both at the
central and peripheral - target - level, namely after binding to its own
receptor. Schistosomin is probably not only responsible for the effects
exerted by the parasite on female reproduction but also for those on male
reproduction and on growth so that energy and space become available for
the continuous production of cercariae. The nature of the humoral cercarial
factor, which induces schistosomin release, is as yet unknown. Based on its
hydrophobic character and on the fact that it can pass through the wall of
the daughter sporocyst, it is supposed to be a diffusible molecule or a
protonephridial excretion product. It does not seem to be a vertebrate-type
steroid, an ecdysteroid or an eicosanoid.
Results obtained in vitro have indicated that schistosomin might have a
suppressive effect on haemocyte activity. Plasma from snails 5-6 weeks
post-exposure showed a tendency to inhibit phagocytic activity of haemo-
cytes from non-infected snails, that is preparatory to the escape and migra-
HOW SCHISTOSOMES PROFITFROM STRESS RESPONSES ELICITEDI N THEIR HOSTS 235
tion of cercariae. Once shedding has started this effect of schistosomin is

overruled by a strong activation of haemocyte activity coinciding with the
tissue damage that the cercariae cause in the host. The cercariae escape from
being attacked by masking their surface coat with host molecules.
As the physiological effects caused by schistosomes resemble those
observed during stress in mammals, experiments were carried out to find
out whether schistosomin is also released in non-parasitized snails during
stress resulting in an inhibiting effect on reproduction. Synergistic action of
short-term stress and other adverse conditions appeared to induce the
release of a factor, probably schistosomin, preventing snails from repro-
ductive activities. So, schistosomin can be compared with, for example, 11-
1, which is released by macrophages during stress in mammals and
transduces signals from the immune system (IS) to the NES. This means
that the effects of schistosome parasites on the NES of the host are
mediated by (factors from) the IDS. Apparently, parasites make use of
existing systems in their host to stop energy-consuming activities, thereby
favouring their own development and reproduction.
In view of these findings about a later stage of infection, it is interesting
to consider the possibility that the inhibiting effects on reproduction in an
early stage of infection of juvenile snails, i.e., on development and
differentiation of the reproductive tract, are also mediated by the IDS. In
this stage of infection activation of haemocytes occurs. This may result in
the release of a second cytokine-like factor, which interferes with the
relevant neuroendocrine regulation mechanisms. Upon penetration into
the snail host miracidia transforming into mother sporocysts release
peptidergic/proteinaceous E/S products modulating the activity of the
IDS of the host. Two main fractions were released in vitro, a 2 kDa
fraction activating haemocyte activity and a 40 kDa fraction suppressing
it. The suppressive factor is supposed to remain close to the surface coat
and to play an important role in compatibility between parasite and host.
The small, activating factor is released and dispersed in the haemolymph of
the snail. In the culture period of 3 4 days, both fractions were released by
the cultured parasites in low and equal amounts explaining that these
fractions when combined no longer had an effect on haemocyte activity.
However, in this period parasites also appeared to induce the host to release
a factor suppressing haemocyte activity, an indirect effect. This host-
derived factor is - as schistosomin - probably also derived from con-
nective tissue cells and/or from haemocytes. If haemocytes are the source
of this factor the production and release of this suppressive factor might
(also) result from their activation by the 2 kDa factor, resembling immuno-
suppression induced by stress in mammals: “the system asks for being
suppressed”. This second cytokine-like factor released by the activated
haemocytes might also be responsible for the effects on the NES resulting
in an inhibited development of the reproductive tract.
Although the 2 kDa and 40 kDa factors in this early stage of infection
have appeared to be peptides/proteins, a role of other E/S products, such as
excretory products and/or diffusible molecules, cannot be excluded. How-
ever, the fact that the effects caused by in vitro released products seem to
reflect the in vivo situation does not favour other possibilities. No indica-
Cytokine-likefactor(s)
schistosomin Neuropeptides
Target tissues
/organs
i L
Physiological
processes
+Effecton ......* } Putative

----+
Produced by --+
Figure 26 Summarizing scheme showing that the effects of the schistosome
parasite Trichobilharzia ocellata on physiological processes in its snail host
Lymnaea stagnalis are stress responses mediated by the internal defence system
(IDS) and the neuroendocrine system (NES). For details see text.
tions were obtained in immunocytochemical studies that the 2 kDa and 40

kDa products represent hormone-like factors. Involvement of heat-shock
proteins in this early stage of infection seems a possibility that is worth
further investigation. Enzymatic degradation of HSP 70 might result in
fragments, respectively activating and suppressing the internal defence in
the host. HSP 70 is not released by exocytosis and therefore cannot be
derived from the secretory vesicles in the (sub)tegumental cells of the
mother sporocyst. Another possibility is that the 40 kDa factor represents
an enzyme as it has been shown that miracidia/mother sporocysts release
enzymes in vitro. In that case these enzymes - as other tegument and
surface coat components - may be derived from the vesicles in the
(sub)tegumental cells or from the apical and lateral glands. The latter
possibility is not likely as it has been shown in in vivo transforming
miracidia that secretion is hampered because the apical papilla, where the
ducts of,these glands end, is retracted and covered by the newly formed
tegument within a few hours after penetration (Meuleman et al., 1978).
Figure 26 summarizes the findings for the model T. ocellata-L. stagnalis.
Because it is not known whether the parasite also has a direct effect on the
NES of the snail host, this possibility has been indicated as “putative”.
Although it has not been demonstrated that neurons, or glial cells, in the
central nervous system of L. stagnalis produce cytokine-like factors, it
seems very likely that this is the case as both neurons (Farrar,1988) and
glial cells (astrocytes, microglia; Benveniste, 1992) of vertebrates have
been shown to produce cytokines. The findings for other parasite-host
combinations, mentioned in this review, show many indications in favour
of the hypothesis that physiological effects caused by endoparasites in their
host are brought about by stress responses they elicit in their host. Experi-
ments on parasite-host interactions mainly focus on the effects of parasitic
infections on either the IDS/IS or the NES. Virtually no attention has been
paid to the effects of parasites on the interactions between these two
regulatory systems and the physiological effects resulting from such inter-
actions. This makes it difficult to determine whether it is a general survival
strategy for parasites to profit from the stress responses they evoke in
their host. As these phenomena are obvious in the schistosome-snail host
model, it seems worthwhile to design experiments aiming at investigating
this hypothesis for other parasite-host partnerships.
ACKNOWLEDGEMENTS
The author is greatly indebted to Dr Elisabeth A. Meuleman for all the

fruitful and indispensable discussions during the preparation of the
manuscript and for critical reading and commenting on it, to all people in
our laboratory who have contributed to the research on parasite-snail
interactions especially Marion Bergamin-Sassen, Robert Hoek and Wessel
Lageweg and to Thea Laan for patience and care in typing the manuscript.
REFERENCES
Abdul-Salam, J.M. and Michelson, E.H. (1980a). Biomphalqria glabrata amoebo-

cytes: effect on in vitro phagocytosis. Journal of Invertebrate Pathology 35,
24 1-248.
Abdul-Salam, J.M. and Michelson, E.H. (1980b). Biomphalaria glabrata amoebo-
cytes: assay of factors influencing in vitro phagocytosis. Journal of Invertebrate
Pathology 36, 52-59.
Adema, C.M., Van der Knaap, W.P.W. and, Sminia, T. (1991a). Molluscan
hemocyte-mediated cytotoxicity: the role of reactive oxygen intermediates.
Reviews in Aquatic Sciences 4, 201-223.
Adema, C.M., Van Deutekom-Mulder, E.C., Van der Knaap, W.P.W., Meuleman,
E.A. and Sminia, T. (1991b). Generation of oxygen radicals in hemocytes of the
snail Lymnaea stagnalis in relation to the rate of phagocytosis. Developmental
and Comparative Immunology 15, 17-26.
Adema, C., Harris, M. and Van Deutekom-Mulder, E.C. (1992). A comparative
study of haemocytes from six different snails: morphology and functional
aspects. Journal of Invertebrate Pathology 59, 24-32.
Alexander, J. and Stimpson, W.H. (1988). Sex hormones and the course of parasitic
infection. Parasitology Today 4, 189-193.
Amen, R.I. and De Jong-Brink, M. (1992). Trichobilharziaocellata in its snail host
Lymnaea stagnalis: an in vitro study showing direct and indirect effects on the
snail’s internal defence system, via the host central nervous system. Parasitology
105, 409-416.
Amen, R.I., Baggen. J.M.C., Bezemer, P.D. and De Jong-Brink, M. (1992). Modu-
lation of the activity of the internal defence system of the pond snail Lymnaea
- stagnalis by the avian schistosome Trichobilharzia ocellata. Parasitology 104,
3340.
Amici, C. and Santoro, G. (1991). Suppression of virus replication by prosta-
glandin A is associated with heat shock protein synthesis. Journal of General
, Virology 72, 1877-1885.
Augustine, O.C. and Denbow, D.M. (1991). Effect of coccidiosis on plasma
epinephrine and norepinephrine levels in turkey poults. Poultry Science 70,
785-789.
Balloul, J.M., Sondermeyer, P., Dreyer, D., Capron, M., Gnych, J.M., Pierce, R.J.,
Carvallo, D., Lecocq, J.P. and Capron, A. (1987). Molecular cloning of a
protective antigen of schistosomes. Nature 326, 149.
Barker, G.C., Chitwood, D.J. and Rees, H.H. (1990). Ecdysteroids in helminths and
annelids. Invertebrate Reproduction and Development 18, 1-1 1.
Bateman,- A., Singh, A., Kral, T. and Solomon, S. (1989). The immuno-
hypothalamic-pituitary-adrenal axis. Endocrine Review 10, 92-1 12.
Bayne, C.J. and Yoshino, T.P. (1989). Determinants of compatibility in molluscs-
trematode parasitism. American Zoologist 29, 399-407.
Bayne, C.J., Buckley, P.M. and Dewan, P.C. (1980a). Macrophage-like haemo-
cytes of resistant Biomphalaria glabrata are cytotoxic for sporocysts of
Schistosoma mansoni in vitro. Journal of Parasitology 66, 4 13-419.
Bayne, C.J., Buckley, P.M. and Dewan, P.C. (1980b). Schistosoma mansoni:
cytotoxicity of haemocytes from susceptible snails for sporocysts in plasma
from resistant Biomphalaria glabrata. Experimental Parasitology 50, 409-416.
Bayomy, M.F.F., Van Elk, R. and Joosse, J. (1989). The effects of infection of
Lymnaea stagnalis with Trichobilharzia ocellata on food consumption and
assimilation, and haemolymph glucose, protein and free amino acid levels.
Proceedings of the Koninklijke Nederlandse Akademie van Wetenschappen
C92, 137-152.
Beck, G. and Habicht, G.S. (1991a). Primitive cytokines: harbingers of vertebrate
defence. Immunology Today 12, 180-188.
Beck, G. and Habicht, G.S. (1991b). Purification and biochemical characterization
of an invertebrate interleukin 1. Molecular Immunology 28, 577-584.
Beck, G., O’Brien, R.F. and Habicht, G.S. (1989). Invertebrate cytokines: the
phylogenetic emergence of interleukin- 1. BioEssays 11, 62-67.
Beckage, N.E. (1993a). Games parasites play: the dynamic roles of proteins and
peptides in the relationship between parasite and host. In “Parasites and
Pathogens of Insects” (N.E. Beckage, S.N. Thompson and B.A. Frederici,
eds), Vol. 1: Parasites, pp. 25-58. Academic Press, New York.
Beckage, N.E. (1993b). Endocrine and neuroendocrine host-parasite relationships.
Receptor 3,233-245.
Beckage, N.E., Zitnan, D. and Sehnal, F. (1994). Endocrine and neuroendocrine
mechanisms of arrested host development in parasitized insects: lessons from the
tobacco hornworm? In “Insect Neurochemistry and Neurophysiology 1993”
(A.B. Borkovec and M.J. Loeb, eds), pp. 123-136. CRC Press, Boca Raton,
Florida.
Becker, W. (1980). Metabolic interrelationships between parasitic trematodes and
molluscs, especially Schistosorna mansoni in Biomphalaria glabrata. Zeitung f i r
Parasitenkunde 634, 101-1 11.
Behnke, J.M. (1990). Evasion of host immunity. In “Parasites: Immunity and
Pathology. The Consequences of Parasitic Infection in Mammals” (J.M.
Behnke, ed.), pp. 344-395. Taylor & Francis, London, New York, Philadelphia.
Benveniste, E.N. (1992). Cytokines: influence on glial cell gene expression and
function. In “Neuroimmunoendocrinology” (J.E. Blalock, ed.), Vol. 52, pp.
106-153. Karger, Basel.
Berkenbosch, F., Van Oers, J., Del Rey, A., Tilders, F. and Besedovsky, H.
( 1987). Corticotropin-releasing-factor-producingneurons in the rat activated by
interleukin 1. Science 238, 524-526.
Beutler, B. and Cerami, A. (1988). Cachectin, cachexia, and shock. Annual
Reviews Medicine 39, 75-83.
Bidmon, H.J. and Koolman, J. (1989). Ecdysteroid receptors located in the central
nervous system of an insect. Experientia 45, 106-109.
Blalock, J.E. (1989). A molecular basis for bidirectional communication between
the immune and neuroendocrine systems. Physiological Reviews 69, 1-32.
Blalock, J.E. (1992). Production of peptide hormones and neurotransmitters by the
immune system. In “Neuroimmunoendocrinology” (J.E. Blalock, ed.), Vol. 52,
pp. 1-24. Karger, Basel.
Blanton, R.E. and Licate, L.S. (1992). Developmental regulation of protein
synthesis in schistosomes. Molecular Biochemical Parasitology 51, 201-208.
240 MARMKE DE JONG-BRINK
Bloom, F.E. (1993). Cell-cell communication in the nervous system. I n “Discus-

sions in Neuroscience” (F.E. Bloom, I.L. Campbell and L. Mucke, eds), Vol.
Molecular and cellular mechanisms of neural-immune interactions., pp. 15-19.
Elsevier, Amsterdam.
Boer, H.H., Van Minnen, J., Stefano, G.B. and Leung, M.K. (1987). Opioid
peptides in the central nervous system (CNS) of Lymnaea stagnalis. In
“Neurobiology: Molluscan Models’’ (H.H. Boer, W.P.M. Geraerts and
J. Joosse, eds), pp. 68-73. North-Holland Publishing, Amsterdam, Oxford, New
York.
Brabin, L. and Brabin, B.J. (1992). Parasitic infection in women and their
consequences. Advances in Parasitology 31, 1-81.
Brooks, M.H., Barry, K.G., Cirksena, W.J., Malloy, J.P., Bruton, J:and Gilliland,
P.F. (1969). Pituitary-adrenal function in acute falciparum malaria. American
Journal of Tropical Medicine and Hygiene 18, 872-877. .
‘Brophy, P.M. and Prichard, D.I. (1992). Immunity to helminths: ready to tip the
biochemical balance? Parasitology Today 8, 419422.
Brown, J.J., Kiuchi, M., Kainoh, Y. and Takeda, S. (1993). In vitro release of
ecdysteroids by an endopiuasitoid, Ascogaster quadrt’dentata Watanabe. Journal
of Insect Physiology 39,229-234.
Bryant, C. (1993). Biochemistry. I n “Modern Parasitology. A Textbook of Para-
sitology” (F.E.G. Cox, ed.), pp. 117-136. Blackwell Scientific Publications,
Oxford.
Buma, P., Roubos, E.W. and Brunekreef, K. (1986). Role of CAMPin electrical and
secretory activity of the neuroendocrine caudo-dorsal cells of Lymnaea stagna-
lis. Brain Research 380,2633.
Bundy, D.A.P. (1988). Gender-dependent patterns of infection and disease. Para-
sitology Today 4, 186-189.
Callahan, H.L., Crouch, R.K. and James, E.R. (1988). Helminth anti-oxidant
enzymes: a protective mechanism against host oxidants? Parasitology Today
4,218-225.
Can, D.J.J. (1992). Neuroendocrine peptide receptors on cells of the immune
system. I n “Neuroimmunoendocrinology” (J.E. Blalock, ed.), Vol. 52, pp.
84-105. Karger, Basel.
Collins, S.M., Hurst, S.M., Stanley, E., Khan, I., Blennerhassett, P. and Swain, M.
(1992). Cytokine-induced changes in neurotransmitter content and release.
Annals of the New York Academy of Sciences 664,415424.
Connors, V.A. and Yoshino, T.P. (1990). In vitro effect of larval Schistosoma
mansoni excretory-secretory products on phagocytosis-stimulated superoxide
production in haemocytes from Biomphalaria glabrata. Journal of Parasitology
76,895-902.
COMOIB,V.A., Lodes, M.J. and Yoshino, T.P. (1991). Identification of a Schisto-
soma mansoni sporocyst excretory-secretory antioxidant molecule and its effect
on superoxide production by Biomphalaria glabrata hemocytes. Journal of
Invertebrate Pathology 58, 387-395.
Cooper, L.A. Ram&, S.K.,Martin, A.E., Richards, C.S. and Lewis, F.A. (1992).
Schistosoma mansoni infections in neonatal Biomphalaria glabrata snails.
Couch, L., Hemi, LA.and Loker, E.S. (1990). Humoral response of the snail
Biomphalaria glabrata to trematode infection: observations on a circulating
hemagglutinin. Journal of Experimental Zoology 255, 34&349.
Cox, F.E.G. (1993). Immunology. I n “Modern Parasitology. A Textbook of Para-
HOW SCHISTOSOMES PROFIT FROM STRESS RESPONSESELICITED IN THEIR HOSTS 241
sitology” (F.E.G. Cox, ed.), pp. 193-218. Blackwell Scientific Publications,

Oxford.
Cox, F.E.G. and Liew, F.Y. (1991). T-cell subsets and cytokines in parasitic
infections. Parasitology Today 8, 371-374.
Crews, A.E., and Yoshino, T.P. (1989). Schistosoma mansoni: effect of infection
on reproduction and gonadal growth in Biomphalaria glabrata. Experimental
Croll, R.P., Van Minnen, J., Smit, A.B. and Kits, K.S. (1991). AGPWamide:
structure, cellular localization and action in Lymnaea stagnalis. In “Molluscan
Neurobiology” (K.S. Kits, H.H. Boer and J. Joosse, eds), pp. 248-254.
North-Holland Publishing, Amsterdam.
Cushing, J.E., Evans, E.E. and Evans, M.L. (1971). Induced bactericidal responses
of abelones. Journal of Invertebrate Pathology 17, 446.
Cutler, L.S. (1983). Cytochemical methods for the localization of adenylate
cyclase. Journal of Histochemistry and Cytochemistry 31, 85-93.
Dahlman, D.L. (1990). Evaluation of teratocyte functions: an overview. Archives of
Insect Biochemistry and Physiology 13, 159-166.
Damian, R.T. (1964). Molecular mimicry: antigen sharing by parasite and host and
its consequences. American Naturalist 98, 120-149.
Damian, R.T. (1987). Molecular mimicry revisited. Parasitology Today 3,
263-266.
Davey, K.G. (1988). Endocrinology of nematodes. In “Endocrinology of Inverte-
brate Types” (H. Laufer, ed.), pp. 63-86. Alan R. Liss, New York.
Davis, T.M.E., Supanaranond, W., Pukrittayakame, S., Krishna, S., Hart, G.R.,
Bumn, J.M., Looareesuwan, S., Vilaiwanna, N. and White, N.J. (1990). The
pituitary-thyroid axis in severe falciparum malaria: evidence for depressed
thyrotroph and thyroid gland function. Transactions of the Royal Sociery of
Tropical Medicine and Hygiene 84, 2330-2334.
De Carvalho, K., Yan, C.Y.I. and De Sousa, W. (1993). Effect of various digestive
enzymes on the interaction of Toxoplasmagondii with macrophages. Parasitology
Research 79, 114-1 18.
De Jong-Brink, M. (1990). How trematode parasites interfere with reproduction of
their invertebrate hosts, freshwater snails. Journal of Medical and Applied
Malacology 2, 101-133.
De Jong-Brink, M. (1992a). Interference of schistosome parasites with neuroendo-
crine mechanisms in their snail host causes physiological changes. Advances in
Neuroimmunology 2, 199-233.
De Jong-Brink, M. (1992b). Neuroendocrine mechanisms underlying the effects of
schistosome parasites on their intermediate snail host. Journal of Invertebrate
Reproduction and Development 22, 127-138.
De Jong-Brink, M. and Bergamin-Sassen, M.J.M. (1989). Trichobilharziaocellata:
influence of infection on the interaction between the dorsal body hormone, a
female gonadotropic hormone, and the follicle cells in the gonad of the
intermediate snail host Lymnaea stagnalis. Experimental Parasitology 68,
93-98.
De Jong-Brink, M. and Geraerts, W.P.M. (1982). Oogenesis in gastropods.
Malacologia 22, 145-149.
De Jong-Brink, M., Elsaadany, M.M., Boer, H.H. and Joosse, J. (1986a). Influence
of trematode parasites upon reproduction activity of their intermediate hosts,
freshwater snail. In “Advances in Invertebrate Reproduction 4” (M. Porchet,
J.C. Andries and A. Dhainaut, eds), pp. 163-172. Elsevier Science B.V.
(Biomedical Division), Amsterdam.
De Jong-Brink, M., Bergamin-Sassen, M.J.M., Kuyt, J.R.M. and Tewari-Kanhai,
A.L. (1986b). Enzyme cytochemical evidence for the activation of adenylate
cyclase in the follicle cells of vitellogenic oocytes by the dorsal body hormone
in the snail Lymnaea stagnalis. General and Comparative Endocrinology 63,
212-219.
De Jong-Brink, M., Elsaadany, M.M. and Boer, H.H. (1988a). Trichobilhurzia
ocellata: interference with the endocrine control of female reproduction of its
host Lymnaea stagnalis. Experimental Parasitology 65, 91-100.
De Jong-Brink, M., Elsaadany, M. and Boer, H.H. (1988b). Schistosomin, an
antagonist of calfluxin. Experimental Parasitology 65, 109-1 18.
De Jong-Brink, M., Schallig, H.F.H., Chatlet, M. and ‘Zonneveld, C. (1989).
. Endocrine interactions between digenetic trematode parasites and their
intermediate hosts, freshwater snails with emphasis on the possible role of
ecdysteroids. Invertebrate Reproduction and Development 15, 202-209.
De Jong-Brink, M., Elsaadany, M. and Solis Soto, .M. (1991). The occurrence of
schistosomin, an antagonist of female gonadotropic hormones, is a general
phenomenon in haemolymph of schistosome-infected freshwater snails.
De Jong-Brink, M., Hordijk, P.L., Vergeest, D.P.E.J., Schallig, H.D.F.H., Kits,
K.S.and Ter Maat, A. (1992).The anti-gonadotropic newpeptide schistosomin
interferes with peripheral and central neuroendocrine mechanisms involved in
the regulation of reproduction and growth in the schistosome-infected snail
Lymnaea stagnalis. In “The Peptidergic Neuron. Rogress in Brain Research”
(J. Joosse, R.M. Buijs and F.J.H. Tilders, eds), Vol. 92,pp. 385-396. Elsevier
Science (Biomedical Division), Amsterdam.
De Jong-Brink, M., Bergamin-Sassen, M.J.M., Broers-Vendrig, T., Glasmeier, A.
and Kits, K.S. (1994). A stress response that delays ovulation egg laying in the
snail Lymnaea stagnalis. Submitted.
Diklcehm, R., Tijnagel, J.M.G.H., Mulder, E.C. and Van der Knaap, W.P.W.
(1987).Haemocytes of the pond snail Lymnaea stagnalis generate reactive forms
of oxygen. Journal of Invertebrate Pathology 49,321-331.
. Dunn,T.S. and Yoshino,T.P. (1988).Schistosoma munsoni: origin and expression
of a tegumental surface antigen on the miracidium and primary sporocysts.
Experimental Parasitology 67, 167-18 1.
Dushay, M.S. and Beckage, N.E. (1993). Dose-dependent separation of Cotesia
’ congregata-associated polydnavirus effects of Manduca sexta larval develop-
ment and immunity.Journal of Insect Physiology 12, 1029-1040.
Duvaux-Miret, 0..Stefano, G.B., Smith, E.M., Dissous, C. and Capron, A. (1992).
Immunosuppression in the definitive and intermediate hosts of the human parasite
Schisrosoma mansoni by release of immunoreactive peptides. Proceedings of the
National Academy of Sciences of the USA 89,778-78 1.
Duvaux-Miret, 0.. Leung, M.K., Capron, A. and Stefano, G.B. (1993).Schistosoma
mansoni: an enkephalonergic system that may participate in internal and
host-ite signalling. Experimental Parasitology 76, 76-84.
Eglezos, A.; Andrews, P.V. and Helme, R.D. (1993).In vivo inhibition of the rat
primary antibody response to antigenic stimulation by somatostatin. Immunology
and Cell Biology 71, 125-129.
Elsasser, T.H., Rumsey, T.S., Hammond, A.C.and Fayer, R. (1988).Influence of
parasitism on plasma concentrations of growth hormone, somatomedin-C and

somatomedin-binding proteins in calves. Journal of Endocrinology 116,
191-200.
Emani, F.P. and Teale, J.M. (1993). Release of stress proteins from Metacestoides
corti is a brefeldin A-inhibitable process: evidence for active export of stress
proteins. Infection and Immunity 61, 2596-2601.
Estes, D,M. and Teale, J.M. (1991). Biochemical and functional analysis of
extracellular stress proteins of Mesocestoides corti. Journal of Immunology
147, 3926-3934.
Farrar, W.L. (1988). Evidence for the common expression of neuroendocrine
hormones and cytokines in the immune and central nervous system. Brain,
Behavior and Immunity 2, 322-327.
Felten, S.Y., Felten, D.L. and Bellinger, D.L. (1992). Noradrenergic and peptider-
gic innervation of the lymphoid organs. In “Neuroimmunoendocrinology” (J.E.
Blalock, ed.), Vol. 52, pp. 25-48. Karger, Basel.
Foster, J.M., Pennock, J.F., Marshall, I. and Rees, H.H. (1993). Biosynthesis of
isoprenoid compounds in Schistosoma mansoni. Molecular and Biochemical
Fryer, S.E. and Bayne, C.J. (1990). Schistosoma mansoni modulation of phago-
cytosis in Biomphalaria glabrata. Journal of Parasitology 76, 45-52.
Fukiishima, T., Isobe, A., Hojo, N., Shiwaku, K., Yamane, Y. and Torii, M. (1993).
The metabolism of arachidonic acid to prostaglandin Ez in plerocercoids of
Spirometra erinacei. Parasitology Research 79, 634-638.
Fusco, A.C., Cassioppi, L., Salafsky, B. and Shibuya, T. (1993). Penetration of
Schistosoma mansoni cercariae into living skin equivalent. Journal of
Gaillard, R.C., Tumill, D., Sappino, P. and Muller, A.F. (1990). Tumor necrosis
factor 01 inhibits the hormonal response of the pituitary gland to hypothalamic
releasing factors. Endocrinology 127.
Gatti, G., Masera, R.G., Pallavicini, L., Sartori, M.L., Stayrenghi, A., Orlandi, F.
and Angeli, A. (1993). Interplay between ACTH, P-endorphin and gluco-
corticoids in the modulation of spontaneous and lymphokine-inducible human
natural killer (NK) cell activity. Brain, Behavior, and Immunity 7, 16-28.
Georgopoulos, C. and Welch, W.J. (1993). Role of the major heat shock proteins as
molecular chaperones. Annual Review of Cellular Biology 9, 601434.
Geraerts, W.P.M. (1976). The role of the lateral lobes in the control of growth and
reproduction in the hermaphrodite freshwater snail Lymnaea stagnalis. General
and Comparative Endocrinology 29, 97-108.
Geraerts, W.P.M. (1992). Neurohormonal control of growth and carbohydrate
metabolism by the light green cell peptides in Lymnaea stagnalis. General
and Comparative Endocrinology 86,433444.
Geraerts, W.P.M. and Joosse, J. (1975). The control of vitellogenesis and of growth
of female accessory sex organs by the dorsal body hormone (DBH) in the
hermaphrodite freshwater snail Lymnaea stagnalis. General and Comparative
Endocrinology 27, 450-467.
Geraerts, W.P.M., Ter Maat, A. and Vreugdenhil, E. (1988). The peptidergic
neuroendocrine control of egg-laying behavior in Aplysia and Lymnaea. In
“Invertebrate Endocrinology” (H. Laufer and R.G.H. Downer, eds), Vol. 2,
Endocrinology of Selected Invertebrate Types, pp. 141-231. Allan R. Liss,
New York.
Geraerts, W.P.M., Smith, A.B., Li, K.W., Vreugdenhil, E. and Van Heerikhuizen, H.
(1991). Neuropeptide gene families that control reproductive behaviour and

growth in molluscs. In “Current Aspects of the Neurosciences” (N.N.
Osbome, ed.), Vol. 3, pp. 255-305. Macmillan, London.
Goudot-Crozei, V., Caillo1.D. Djabali, M. and Dessein, A.J. (1989). The major
parasite surface antigen associated with human resistance to schistosomiasis is a
37-kDa glyceraldehyde-3-phosphatedehydrogenase. Journal of Experimental
Medicine 170, 2065-2080.
Granath, W.O.and Yoshino, T.P. (1983). Lysosomal enzyme activities in sus-
ceptible and refractory strains of Biomphalaria glabrata during the course of
infection with Schistosoma mansoni. Journal of Parasitology 69, 1018-1026.
Granath, W.O. and Yoshino, T.P. (1984). Schistosoma mansoni: passive transfer of
resistance by serum in the vector snail Biomphalaria glabrata. Experimental
Haas, W. (1992). Physiological analysis of cercarial behavior. Journal of
* Parasitology 78, 243-255.
Haas, W. and Voigt, W.P. (1988). Host finding - A physiological effect. In
“Parasitology in Focus. Facts and Trends”. (H. Mehbom, ed), pp. 454-464.
Springer-Verlag, Berlin, Heidelberg, New York, London, Paris, Tokyo.
Ham, D.A., Gu, W., Oligino, L.D., Mitsuyama, M., Gebremichael, A. and Richter,
D. (1992). A protective monoclonal antibody specifically recognizes and alters
the catalytic activity of schistosome triose-phosphate isomerase. Journal of
Immunology 148,562-567.
Hayakawa, Y. (1992). A putative new juvenile peptide hormone in lepidopteran
insects. Biochemical Biophysical Research Communications 185, 1141-1 147.
Hemminga, M.A., Maaskant, J.J. and Joosse, J. (1985). Direct effects of the
hyperglycaemic factor of the freshwater snail Lymnaea stagnalis on isolated
glycogen cells. General and Comparative Endocrinology 58, 131-1 36.
Hoek, R.M., Smit, A.B., Van der Sar, A., Lageweg, W.,Geraerts, W.P.M. and De
Jong-Brink, M. (1994). Schistosomin, a novel cytokine-like molecule from
mollusc. Lymnaea stagnalis. Submitted.
Hong, Z., Kosman, D.J., Thakur, A., Rekosh, D. and LoVerde, P.T. (1992).
Identification and purification of a second form of CWZn superoxide dismutase
from Schistosoma mansoni. Infection and Immunity, 3641-365 1.
Hong, Z., LoVerde, P.T., Thakur, A., Hammarskjold, M.L. and Rekosh, D. (1993).
.Schistosoma mansoni: a CrJZn superoxide dismutase is glycosylated when
expressed in mammalian cells and localizes to a subtegumental region in adult
schistosomes. Experimental Parasitology 76, 101-1 14.
Hordijk, P., Van Loenhout, H., Ebberink, R.H.M., De Jong-Brink, M. and Joosse, J.
(1991a). Neuropeptide schistosomin inhibits hormonally-induced ovulation in
the freshwater snail Lymnaea stagnalis. Journal of Experimental Zoology 259,
268-271.
Hordijk, P.L., Ebberink, R.H.M., De Jong-Brink, M. and Joosse, J. (1991b).
Isolation of schistosomin, a neuropeptide which antagonizes gonadotropic
hormones in a freshwater snail. European Journal of Biochemistry 195,131-136.
Hordijk, P.L., Schallig, H.D.F.H., Ebberink, R.H.M., De Jong-Brink, M. and
Joosse, J. (1991~).Primary structure and origin of schistosomin, an anti-
gonadotropic neuropeptide of the pond snail Lymnaea stagnalis. Biochemical
Journal 279,-837-842.
Hordijk, P.L., Ebberink, R.H.M., De Jong-Brink, M. and Joosse, J. (1991d).
Purification and characterization of the neuropeptide schistosomin which inhi-
bits the bioactivity of gonadotropic hormones in trematode-infected Lymnaea
stagnalis. In “Molluscan Neurobiology” (K.S. Kits, H.H. Boer and J. Joosse,

eds), pp. 340-345. North-Holland Publishing, Amsterdam.
Hordijk, P.L., De Jong-Brink, M., Lodder, J.C., Pieneman, A.B., Ter Maat, A. and
Kits, K.S. (1992). The neuropeptide schistosomin and haemolymph from para-
sitized snails induce similar changes in excitability in neuroendocrine cells
controlling reproduction and growth in a freshwater snail. Neuroscience Letters
136, 193-197.
Horta, M.F.M. and Ramalho-Pinto, F.J. (1991). Role of human decay-accelerating
factor in the evasion of Schistosoma mansoni from the complement-killing in
vitro. Journal of Experimental Medicine 174, 1399-1406.
Hughes, T.K., Smith, E.M., Chin, R., Cadet, P., Sinisterra, J., Leung, M.K., Shipp,
M.A., Scharrer, B. and Stefano, G.B. (1990). Interaction of immunoactive
monkines (interleukin I and tumor necrosis factor) in the bivalve mollusc
Mytilus edulis. Proceedings of the National Academy of Sciences of the USA
87,44264429.
Hurd, H. (1990). Physiological and behavioural interactions between parasites and
invertebrate hosts. Advances in Parasitology 29, 271-318.
Hurd, H. (1993). Reproductive disturbances induced by parasites and pathogens of
insects. In “Parasites and Pathogens of Insects” (N.E. Beckage, S.N. Thompson
and B.A. Frederici, eds), Vol. 1, Parasites, pp. 87-105. Academic Press, New
York.
Imura, H., Fukata, J. and Mori, T. (1991). Cytokines and endocrine function: an
interaction between the immune and neuroendocrine systems. Clinical
Endocrinology 35, 107-1 15.
Isseroff, H., Sylvester, P.W. and Held, W.A. (1986). Effects of Schistosoma
mansoni on androgen regulated gene expression in the mouse. Molecular
Biochemistry and Parasitology 18, 401412.
Isseroff, H., Sylvester, P.W., Bessette, C.L., Jones, P.L., Fisher, W.G., Rynkowski,
T.A. and Gregor, K.R. (1989). Schistosomiasis: role of endogenous opioids in
suppression of gonadal steroid secretion. Comparative Biochemical Physiology
94A, 4 1 4 5 .
Johnson, H.M., Downs, M.O. and Pontzer, C.H. (1992). Neuroendocrine peptide
hormone regulation of immunity. In “Neuroimmunoendocrinology” (J.E.
Blalock, ed.), Vol. 52, pp. 49-83. Karger, Basel.
Joosse, J. (1984). Recent progress in endocrinology of molluscs. In “Biosynthesis,
Metabolism and Mode of Action of Invertebrate Hormones” (J. Hoffmann and
M. Porchet, eds), pp. 19-35. Springer, Berlin, Heidelberg, New York.
Joosse, J. (1988). The hormones of molluscs. In “Invertebrate Endocrinology”
(H.Laufer and R.G.H. Downer, eds), Vol. 2. Endocrinology of Selected
Invertebrate Types, pp. 89-140. Alan Liss, New York.
Joosse, J. and Van Elk, R. (1986). Trichobilharzia ocellata: physiological
characterization of giant growth, glycogen depletion, and absence of reproduc-
tive activity in the intermediate snail host, Lymnaea stangalis. Experimental
Joosse, J., Van Elk, R., Mosselman, H. and Wortelboer, H. and Van Diepen, J.C.E.
(1988). Schistosomin: a pronase-sensitive agent in the haemolymph of Tricho-
bilharzia ocellata-infected Lymnaea stagnalis inhibits the activity of albumen
glands in vitro. Parasitology Research 74, 228-234.
Joplin, K.H. Yocum, G.D. and Denlinger, D.L. (1990). Cold shock elicits expres-
sion of heat shock proteins in the fresh fly, Drosophila crassipalpis. Journal of
Insect Physiology 36, 825-834.
Kaufmann, S.H.E. (1990a). Heat shock proteins and the immune response.
Immunology Today 11, 129-136.
Kaufmann, S.H.E. (1990b). Heat-shock proteins: a missing link in the host-parasite
relationship? Medical Microbiology and Immunology 179, 6 1 4 6 .
Kavaliers, M. and Colwell, D.D. (1992). Parasitism, opioid systems and host
behaviour. Advances in Neuroimmumlogy 2, 287-295.
Kavaliers, M. and Podesta, R.B. (1988). Opioid involvement in parasite-induced
behavioural modifications: evidence from hamsters infected with Schistosoma
mansoni. Canadian Journal of Zoology 66, 2653-2657.
Kavaliers, M., Podesta, R.B., Hirst, M. and Young, B. (1984). Evidence for the
activation of the endogenous opiate system in hamsters infected with human
blood flukes. Life Sciences 35, 2365-2373.
Kavelaars, A., Ballieux, R.E. and Heynen, C.I. (1989). The role of I11 in the
corticotropin release factor and arginine-induced vasopressin-induced secretion
of immunoreactive beta-endorphin by human peripheral blood mononuclear
cells. Journal of Immunology 342,2338-2342.
Kehoe, J.S. (1975). Electrogenic effects of neutral ,amino acids on neurons of
Aplysia californica. Cold Spring Harbor Symposium, The Synaps 40, 145-155.
Kitano, H., Wago, H. and Arakawa, T. (1990). Possible role of teratocytes of the
gregarious parasitoid, Cotesis ( = Apanteles) glomerata in the suppression of
phenoloxidase activity in the larval host, Pieris rapae crucivora. Archives of
Insect Biochemistry and Physiology 13, 177-1 85.
Kits, K.S. (1980). States of excitability in ovulation hormone producing neuro-
endocrine cells of Lymnaea stagnalis (Gastropoda) in their relation to the
egg-laying cycle. Journal of Neurobiology 11, 397410.
K(lhler, P. and Voigt, W.P. (1988). Nutrition and metabolism. In “Parasitology in
Focus. Facts and Trends” (H. Mehlhorn, ed.), pp. 412453. Springer-Verlag,
Berlin, Heidelberg, New York, London, Paris, Tokyo.
Koolman, J. (1990). The occurrence of ecdysteroids in vertebrates infected with
helminths. Progress in Clinical and Biological Research 342, 704-709.
Kuchler, K. (1993). Unusual routes of protein secretion: the easy way out. Trends
in Cell Biology 3, 421426.
Langley, J.G. and Dunne, D.W. (1992). Temporal variation in the carbohydrate and
peptide surface epitopes in antibody-dependent, eosinophil-mediated killing of
-Schistosoma mansoni schistosomula. Parasite Immunology 14, 185-200.
Lawrence, P.O. (1990). Serosal cells of Biosreres longicaudarus (Hymenoptera:
Braconidae): ultrastructure and release of polypeptides. Archives of Insect
Biochemical Physiology 13, 199-216.
Lawrence, P.O. and Lanzrein, B. (1993). Hormonal interactions between insect
endoparasites and their host insects. In “Parasites and Pathogens of Insects”
(N.E. Beckage, S.N. Thompson and B.A. Frederici, eds), Vol. 1: Parasites, pp.
59-86. Academic Press, New York.
k h a n , R.M., Toni, R., Clark, B.D., Cannon, J.G., Shaw, A.R., Dinarelo, C.A.
and Reichlin, S. (1990). Immunoreactive interleukin-1f3 localization in the rat
forebrain. Brain Research 514, 135-140.
Lie, K.J. (1982). Survival of Schistosoma mansoni and other trematode larvae in
the snail Bbmphalaria glabrara. A discussion of the interference theory.
Tropical and Geographicat Medicine 34, 111-122.
Lie, K.J., Jeong, K.H. and Heyneman, D. (1987). Molluscan host reactions to
helminthic infection. In “Immunology, Immunopathology, and Immunopro-
phylaxis” (E.J.L. Soulsby, ed.), Vol. IV, pp. 211-270. CRC Press, Boca
Raton, Florida.
Liew, F.Y. and Cox, F.E.G. (1991). Nonspecific defence mechanisms: the role of
nitric oxide. In Immunoparasitology Today (Immunology Today 12, Parasitology
Today 7), A17-A21. Elsevier Trends Journals, Cambridge.
Lightowlers, M.W. and Rickard, M.D. (1988). Excretory-secretory products of
helminth parasites: effect on host immune responses. Parasitology 96, 123-166.
Lindquist, S. and Craig, E.A. (1988). The heatshock proteins. Annual Reviews of
Genetics 22, 631-677.
Lingelbach, K.R. (1993). Plasmodium falciparum: a molecular view of protein
transport from the parasite into the host erythrocyte. Experimental Parasitology
76, 318-327.
Lodes, M.J. and Yoshino, T.P. (1990). The effect of schistosome excretory-
secretory products on Biomphalaria glabrata haemocyte motility. Journal of
Invertebrate Pathology 56, 75-85.
Lodes, M.J., Connors, V.A. and Yoshino, T.P. (1991). Isolation and functional
characterization of snail hemocyte-modulating polypeptide from primary sporo-
cysts of Schistosoma mansoni. Molecular and Biochemical Parasitology 49,
1-10.
Loker, E.S. and Bayne, C.J. (1982). In vitro encounters between Schistosoma
mansoni sporocysts and hemolymph components of susceptible and resistant
strains of Biomphalaria glabrata. American Journal of Tropical Medicine and
Hygiene 31,999-1005.
Loker, E.S. and Bayne, C.J. (1986). Immunity to trematode larvae in the snail
Biomphalaria. Symposia Zoological Society of London 56, 199-220.
Loker, E.S., Bayne, C.J. and Yui, M.A. (1986). Echinostoma paraensei: hemocytes
of Biomphalaria glabrata as targets of echinostome mediated interference with
snail host resistance to Schistosoma mansoni. Experimental Parasitology 62,
149-154.
Loker, E.S., Cimino, D.F. and Hertel, L.A. (1992). Excretory-secretory products of
Echinostoma paraensei sporocysts mediate interference with Biomphalaria
glabrata hemocyte functions. Journal of Parasitology 73, 1090-1098.
Lolait, S.J., Clements, J.A., Marck-Wick, A.J., Chang, C., McNally, M., Smith,
A.J. and Funder, J.W. (1986). Proopiomelanocortin messenger ribonucleic acid
and post-translational processing of beta-endorphin in spleen macrophages.
Journal of Clinical Investigation 77, 1776-1779.
Maizels, R.M., Bundy, D.A.P., Selkirk, M.E., Smith, D.F. and Anderson, R.M.
(1993). Immunological modulation and evasion by helminth parasites in human
populations. Nature 365, 797-805.
Maresca, B. and Caratu, L. (1992). The biology of the heat shock response in
parasites. Parasitology Today 8, 260-266.
McClelland, G. and Bourns, T.K.R. (1969). Effects of Trichobilharzia ocellata
on growth, reproduction and survival of Lymnaea stagnalis. Experimental
Parasitology 24, 137- 146.
McKerrow, J.H. and Doenhoff, M.J. (1988). Schistosome proteases. Parasitology
Today 4, 334-340.
McLaren, D.J. (1980). Schistosoma mansoni: the parasite surface in relation to host
immunity. In “Tropical Medicine Research Studies” (K.H. Brown, ed.),
Research Studies Press, Chichester.
Meier, M. and Meier-Brook, C. (1981). Schistosoma mansoni: effect on growth,
fertility, and development of distale male organs in Biomphalaria glabrata

exposed to miracidia at different ages. Zeitung fur Parasitenkunde 66, 121-131.
Meuleman, E.A. (1972). Host-parasite interrelationships between the freshwater
pulmonate Biomphalaria -pfeifferi and the trematode Schistosoma mansoni.
Netherlands Journal of Zoology 22, 355427.
Meuleman, E.A. and Holzmann, P.J. (1975). The development of the primitive
epithelium and true tegument in the cercaria oE Sehistosoma mansoni. Zeitung
f i r Parasitenkunde 45, 307-3 18.
Meuleman, E.A., Lyaruu, D.M., Khan, M.A., Holzmann, P.J. and Sminia, T.
(1978). Ultrastructural changes in the body wall of Schistosoma mansoni during
the transformation of the miracidum into the mother sporocyst in the snail host
Biomphalaria pfeireri. Zeitung fiir Parasitenkunde 56, 227-2422
Meuleman, E.A., Holzmann, P.J. and Peet, R.C. (1980). The development of
daughter sporocysts inside the mother sporocyst of Schistosoma mansoni with
special reference to the ultrastructure of the body wall. Zeitschrijl fur
Meuleman, E.A., Huyer, A.R. and Mooij, J.H. (1984). Maintenance of the life
cycle of Trichobilharzia ocellata via the duck Anas platyrhynchos and the pond
snail Lymnaea stagnalis. Netherlands Journal of Zoology 34, 414417.
Mitchell, G.F. (1989). Glutathione-S-transferases-potential components of anti-
schistosome vaccines? Parasitology Today 5, 34-37.
Minchilla, D.J. and Loverde, P.T. (1981). A cost of increased early reproductive
effort in the snail Biomphalaria glabrata. American Naturalist 118, 876-881.
Moed, P.J., Pieneman, A.W., Bos. N.P. and Ter Maat, A. (1989). The role of CAMP
in regulation of electrical activity of the neuroendocrine caudodorsal cells of
Lymnaea stagnalis. Brain Research 476, 298-306.
Mohrig, W. and Shittek, D. (1979). Phagocytosis stimulating mediators in insects.
Acta Biologica of Medica Germanica 38, 953-958.
MONOY,F.P. and Loker, E.S.(1993). Production of heterogeneous carbohydrate-
binding proteins by the host snail Biomphalaria glabrata following exposure to
Echinostoma paraensei and Schistosoma mansoni. Journal of Parasitology 79,
41-23.
Momson, D.D., Waa, E.A. and Bennet, J.L. (1986). Effects of steroids and steroid
synthesis inhibitors on fecundity of Schistosoma mansoni in vitro. Journal of
.Chemical Ecology 12, 1901-1908.
Nair, M.P.N., Schwartz, S.A., Wu, K. and Kronfol, 2.(1993). Effect of neuro-
peptide Y on natural killer activity of normal human lymphocytes. Brain,
Behavior and Immunity 7 , 7 6 7 8 .
Nafe, B., Smith, J.M. and Prichard, R.K. (1990). Schistosoma mansoni: levels of
antioxidants and resistance to oxidants increase during development.
Experimental Parasitology 70, 389-397.
Nare, B., Smith, J.M. and Prichard, R.K. (1991). Oltipraz-induced decrease in
the activity of cytosolic glutathione S-transferase in Schistosoma mansoni.
International Journal of Parasitology 21, 9 19-925.
Nare, B., Smith, J.M. and Prichard, R.K. (1992). Mechanisms of inactivation of
Schistosoma mansoni and mammalian glutathione S-transferase activity by the
antischistosomal drug oltipraz. Biochemical Pharmacology 43, 1345-1 35 1.
Navarra, P.. Tsagarakis, S.,Fiara, M.S., Rees, L.H., Besser, G.M. and Grossmann,
A.B. (1991). Interleukins-1 and -6 stimulate the release of corticotropin-releasing
hormone41 from rat hypothalamus in vitro via the eicosanoid cyclooxygenase
pathway. Endocrinology 1 2 8 , 3 7 4 .
Neumann, S., Ziv, E., Lantner, F. and Schlechter, I. (1993). Regulation of HSP70
gene expression during the life cycle of the parasitic helminth Schistosoma
mansoni. European Journal of Biochemistry 212, 589-596.
Nevhutalic, P.A., Salafsky, B., Haas, W. and Conway, T. (1993). Schistosoma
mansoni and Trichobilharzia ocellata: comparison of secreted cercarial
eicosanoids. Journal of Parasitology 79, 130-1 33.
Newport, G., Culpepper, J. and Agabian, N. (1988). Parasite heat-shock proteins.
Parasitology Today 4, 306-3 12.
Nicaise, G. (1973). The gliointerstitial system of molluscs. In “International
Review of Cytology” (G.H. Bourne, J.F. Danielli and K.W. Jeon, eds), Vol.
34, pp. 251-332. Academic Press, New York, San Francisco, London.
Nilsson, L.A., Lange, S. and Lonnroth, I. (1992). Induction of anti-secretory factor
in mice by the nonintestinal parasite Schistosoma mansoni. Journal of Para-
sitology 78, 1055-1058.
Nird6, P., Torpier, G., Capron, A., Delaage, M. and De Reggi, M.L. (1984).
Ecdysteroids in schistosomes and host-parasite relationships. In “Biosynthesis,
Metabolism and Mode of Action of Invertebrate Hormones” (J.A. Hoffmann and
M. Porchet, eds). Springer-Verlag, Berlin.
Nisticb, G. (1993). Communications among central nervous system, neuro-
endocrine and immune systems: interleukin-2. Progress in Neurobiology 40,
463475.
Noda, S. and Loker, E.S. (1989). Phagocytotic activity of hemocytes of M-line
Biomphalaria glabrata snails: effect of exposure to the trematode Echinostoma
paraensei. Journal of Parasitology 75, 26 1-269.
Nozaki, T. and Dvorak, J.A. (1993). Intraspecific diversity in the response of
Trypanosomacruzi to environmental stress. Journal of Parasitology 79.45 1454.
Ndiiez, P.E. and De Jong-Brink, M. (1994). The suppressive excretory-secretory
product of Trichobilharzia ocellata: a possible factor for determining com-
patibility in parasite-host interactions. Journal of Parasitol. Research.
Submitted.
Nliiiez, P.E., Adema, C.M. and De Jong-Brink, M. (1994a). Modulation of the
killing activity of haemocytes of Lymnaea stagnalis by the trematode parasite
Trichobilharzia ocellata. Parasitology 109, 299-3 10.
Nliiiez, P.E.,Li, K.W. and De Jong-Brink, M. (1994b). Excretory-secretory pro-
ducts of Trichobilharzia ocellata and their modulating effects on the internal
defence system of Lymnaea stagnalis. Submitted.
Oehlmann, J. and Bettin, C. (1992). TBT-induced imposex and the role of steroids
in marine snails. In “Abstracts 11 International Malacological Congress”
(F. Giusti and G. Manganelli, eds). University of Siena, Siena.
Ottaviani, E., Petraglia, F., Montagnani, G., Cossarizza, A., Monti, D. and
Franceschi, C. ( 1990). Presence of ACTH- and P-endorphin-immunoreactive
molecules in the freshwater snail Planorbarius corneus (L.) (Gastropoda,
Pulmonata). Regulatory Peptides 27, 1-9.
Ottaviani, E., Caselgrandi, E., Petraglia, F. and Franceschi, C. (1992). Stress
response in the freshwater snail Planorbarius corneus (L.) (Gastropoda, Pulmo-
nata): interaction between CRF, ACTH, and biogenic amines. General and
Comparative Endocrinology 87, 354-360.
Ottaviani, E., Caselgrandi, E., Franchini, A. and Franceschi, C. (1993a). CRF
provokes the release of norepinephrine by hemocytes of Viviparis ater (Gastro-
poda, Prosobranchia): further evidence in favour of the evolutionary hypothesis
of the “mobile immune-brain”. Biochemical and Biophysical Research

Communications 193, 446-452.
Ottaviani, E., Franchini, A. and Franceschi, C. (1993b). Presence of several
cytokine-like molecules in molluscan hemocytes. Biochemical and Biophysical
Research Communications 195, 984-988.
Pan, C.T. (1965). Studies on the host-parasite relationship between Schistosoma
mansoni and the snail Australorbis glabrata. American Journal of Tropical
Medicine and Hygiene 14, 93 1-976.
Pang, G., Coach, L., Batey, R., Clancy, R. and Cripss, A. (1994). GM-CSF, 11-1-a,
11-1-p, 11-6, 11-8, 11-10, ICam-1 and VCam-1 gene expression and cytokine
production in human duodenal fibroblasts stimulated with lipopolysaccharide,
11-1-a and TNF-a. Clinical and Experimental Immunology 96, 437443.
Payan, D.G., Hess, C.A. and Goetzl, E.J. (1984). Inhibition by somatostatin of the
proliferation of T lymphocytes and Molt4 lymphoblasts. Cellular Immunology
84,433438.
Pearce, E.J., Hall, F. and Sher, A. (1990). Host-specific evasion of the alternate
complement pathway by schistosomes correlates with the presence of phospho-
lipase C-sensitive surface molecule resembling human decay accelerating factor.
Journal of Immunology 144, 2751-2756.
Pentreath, V.W., Radojcic, T., Seal, L.H. and Winstanley, E.K. (1985). The glial
cells and glia-neuron relations in the bucca ganglia of Planorbis corneus (L.):
cytological qualitative and quantitative changes during growth and ageing.
Philosophical Transactions of the Royal Society of London B 307, 399455.
Phares, C.K. (1988). Use of receptor affinity chromatography in purification of
the growth hormone-like factor produced by plerocercoids of the tapeworm
Spirometra mansonoides. Journal of Receptor Research 8, 645-665.
Phares (1992). Biological characteristics of the growth hormone-like factor from
plerocercoids of the tapeworm Spirometra mansonoides. Advances in Neuro-
immunology 2, 235-247.
Phares, C.K. and Watts, D.J. (1988). The growth hormone-like factor produced by
the tapeworm Spirometra mansonoides specifically binds receptors on cultured
human lymphocytes. Journal of Parasitology 74, 896-898.
Phares, C.K., Schafer, J.L. and Heidrick, M.L. (1990). Characteristics, distribution,
and possible evolutionary importance of the human growth hormone-like factor
.from plerocercoids of the tapeworm genus Spirometra. In “Immune Recognition
and Evasion: Molecular Aspects of Host-Parasite Interaction” pp. 149-161.
Academic Press, Inc.
Polla, B.S. (1991). Heat shock proteins in host-parasite interactions. Immuno-
parasitology Today A38-A41.
Raftos, D.A., Cooper, E.L., Habicht, G.S. and Beck, G. (1991). Invertebrate cyto-
kines: tunicate cell proliferation stimulated by an interleukin 1-like molecule.
Proceedings of the National Academy of Sciences of the USA 88, 95 18-9522.
Ratner, S. and Vinson, S.B. (1983). Phagocytosis and encapsulation: cellular
immune responses in arthropoda. American Zoologists 23, 185-194.
Reincke, M., Heppner, C., Petzke, F., Allolio, B., Arlt, W., Mbulamberi, D.,
Siekmann, L., Vollmer, D., Winkelmann, W. and Chrousos, G.P. (1994). Impair-
ment of adrenocortical function associated with increased plasma tumor
necrosis factor-a and interleukin-6 concentrations in African trypanosomya-
sis. Neuroimmunomodulation 1, 14-22.
Riley, E.M. and Chappell, L.H. (1990). Praziquantel treatment of Biomphalaria
glabrata infected with Schistosoma mansoni - influence on snail fecundity.

Roitt, I., Brostoff, J. and Male, D. (1993). “Immunology.” Mosby-Year Book
Europe Limited, London.
Roszman, T.L. and Brooks, W. (1992). Signalling pathways of the neuroendocrine-
immune network. In “Neuroimmunoendocrinology” (J.E. Blalock, ed.), Vol. 52,
pp. 17&190. Karger, Basel.
Rothwell, N. (1991). Functions and mechanisms of interleukin 1 in the brain.
Trends in Pharmacological Sciences 121, 430-436.
Rubiliahi, C. (1983). Action of rhizocephalan on the genital activity of host male
crabs: characterization of a parasitic secretion inhibiting spermatogenesis.
International Journal of Invertebrate Reproduction 6 , 137-147.
Rubiliani, C. and Payen, G. (1988). Les sacculines: d’etranges parasites castrateurs.
Herssey, Evreux 45171, 193-200.
Salafsky, B. and Fusco, A. (1987). Eicosanoids as immunomodulators of penetration
by schistosome cercariae. Parasitology Today 3, 279-281.
Sapolsky, R.M. (1992). Neuroendocrinology of the stress-response. In “Behavioral
Endocrinology” (J.B. Becker, S.M. Breedlove and D. Crews, eds), pp. 287-324.
Massachusetts Institute of Technology. A Bradford Book, MIT Press,
Cambridge, Massachusetts; London, England.
Sapolsky, R., Rivier, C., Yamamoto, G., Plotsky, P. and Val, W. (1987). Interleukin-
1 stimulates the secretion of hypothalamic corticotropin releasing factor. Science
238,522-524.
Schallig, H.D.F.H. (1991). Neuroendocrine interactions between schistosome
parasites and their intermediate hosts, freshwater snails, studied in the combi-
nation Trichobilharzia ocellata-Lymnaea stagnalis. Thesis Vrije Universiteit
Amsterdam. pp. 95-103. Febodruk, Enschede.
Schallig, H.D.F.H., Young, N.J., Magee, R.M., de Jong-Brink, M. and Rees, H.H.
(1991a). Identification of free and conjugated ecdysteroids in cercariae of the
schistosome Trichobilharzia ocellata. Molecular and Biochemical Parasitology
49, 169-176.
Schallig, H.D.F.H., Hordijk, P.L., Oosthoek, P.W. and De Jong-Brink, M. (1991b).
Schistosomin, a peptide present in the haemolymph of Lymnaea stagnalis
infected with Trichobilharzia ocellata, is only produced in the snail’s central
nervous system. Parasitology Research 77, 152-156.
Schallig, H.D.F.H., Sassen, M.J.M., Hordijk, P.L. and De Jong-Brink, M. (1991~).
Trichobilharzia ocellata infection influence on the fecundity of its intermediate
snail host Lymnaea stagnalis and cercarial induction of the release of schisto-
somin, a snail neuropeptide antagonizing female gonadotropic hormones.
Schallig, H.D.F.H., Glasmeier, A. and De Jong-Brink, M. (1992a). Vertebrate type
steroids in cercariae of the schistosome Trichobilharzia ocellata. Parasitology
Research 78, 709-7 11.
Schallig, H.D.F.H., Sassen, M.J.M. and De Jong-Brink, M. (1992b). In vifrorelease
of the anti-gonadotropic hormone, schistosomin, from the central nervous system
of Lymnaea stagnalis is induced with a methanolic extract of cercariae of
Trichobilharzia ocellata. Parasitology 104, 309-3 14.
Schutt, D.A., Smith, A.I., Wallace, C.A., Connell, R. and Fell, L.R. (1988). Effect
of myiasis and acute restraint stress on plasma levels of immunoreactive beta-
endorphin, adrenocorticotrophin (ACTH) and cortisol in the sheep. Australian
Journal of Biological Sciences 41, 297-301.
Segura, J.J., Guerrero, J.M., Goberna, R. and Calvo, J.R. (1992). Somatostatin
inhibition of VIP- and isoproterenol-stimulated cyclic-AMP production in rat
peritoneal macrophages. Neuropeptides 23, 2 9 4 3 .
Shiff, C.J. and Dossaji, S.F. (1991). Ecdysteroids as regulators of host and parasite
interactions: a study of interrelationships between Schistosoma mansoni and the
host snail Biomphalaria glabrata. Tropical Medical Parasitology 42, 11-1 6.
Shinnick, T.M. (1991). Heat shock proteins as antigens of bacterial and parasitic
pathogens. In “Current Topics in Microbiology and Immunology” (S.H.E.
Kaufmann, ed.), Vol. 167: Heat shock proteins and immune responses, pp.
145-160. Springer-Verlag, Berlin.
Shoemaker, G.B., Ramachandran, H., Landa, A., dos Reis, M.G. and Stein, L.D.
(1992). Alternative splicing of the Schistosoma mansoni gene encoding a
homologue of epidermal growth factor receptor. Molecular and Biochemical
’Shoemaker, C.B., Reynolds, S.R., Wei, G., Tielens, A.G.M. and Ham, D.A. (1994).
Schistosoma mansoni hexokinase: cDNA cloning and immunogenecity studies.
Experimental Parasitology. In press.
Simpson, A.J.G. and Smithers, S.R. (1985). Schistosomes: surface, egg and circu-
lating antigens. Current topics in Microbiology and Immunology 120, 205-223.
Sluiters, J.F. (1981). Development of Trichobilharzia ocellata in Lymnaea stag-
nalis and the effects of infection on the reproductive system of the host. Zeitung
fur Parasitenkunde 64, 303-3 19.
Sluiters, J.F., Brussard-Wust, C.M. and Meuleman, E.A. (1980). The relationship
between miracidial dose, production of cercariae, and reproductive activity of
the host in the combination Trichobilharzia ocellata and Lymnaea stagnalis.
Zeitschrqt fur Parasitenkunde 63, 13-26.
Sminia, T.S. (1972). Structure and function of blood and connective tissue cells of
the freshwater pulmonate Lymnaea stagnalis studied by electron microscopy and
enzyme histochemistry. Zeitung fur Zellforschung 130,497-526.
Sminia, T.S.and Van der Knaap, W.P.W. (1981). The internal defence system of
the freshwater snail Lymnaea stagnalis. Developmental and Comparative
Immunology 5, 87-97.
Sminia, T.S.and Van der Knaap, W.P.W. (1986). Immunorecognition in inverte-
brates with special reference to molluscs. In “Immunity in Invertebrates”
(M. Brehelin, ed.), pp. 112-124. Springer-Verlag, Berlin-Heidelberg.
Sminia, T.S., Borghart-Reinders, E. and Van der Linde, A.W. (1974). Encapsula-
tion of foreign material experimentally introduced into the freshwater snail
Lymnaea stagnalis. Cell and Tissue Research 153, 307-326.
Smjt, A.B., Vreugdenhil, E., Ebberink, R.H.M., Geraerts, W.P.M., Klootwijk, J. and
Joosse, J. (1988). Growth controlling molluscan neurons contain the precursor of
molluscan insulin-related peptide. Nature 331, 535-538.
Smith, E.M. (1989). Neuropeptide gene expression in lymphoid tissue. In
“Endogenous Sleep Factors” (S. Inoue and J.M. Krueger, eds), Bouma, Text,
Wassenaar.
Smith, E.M. and Johnson, E.W. (1991). Neuropeptides and neuropeptide receptors
in the immune system. In “Studies in Neuroscience. Comparative Aspects of
Neuropeptide Function” (E. Florey and G.B. Stefano, eds), pp. 3 13-316.
Manchester University Press, Manchester.
Soderhall, K. and A s p h , A. (1993). Prophenol oxidase activity system and its
role in cellular communication. In “Insect Immunity” (J.P.N. Pathak, ed.), pp.
113-130. Kluwer Academic, Dordrecht, Boston, London.
Solis-Soto, M. and De Jong-Brink, M. (1994). Immunocytochemical study on

biologically active neurosubstances in daughter sporocysts and cercariae of
Trichobilharzia ocellata and Schistosoma mansoni. Parasitology 108, 301-3 1 1.
Soudan, B., Tetaert, D., Racadot, A., Degand, P. and Boersma, A. (1992). Decrease
of testoster~ne level during an experimental African trypanosomiasis. Acta
Endocrinologica Copenhagen 127, 86-92.
Srivastava, P.K. and Maki, R.G., (1991). Stress-induced proteins in immune
responses. In “Current Topics in Microbiology and Immunology” (S.H.E.
Kaufmann, ed.), Vol. 167: Heat shock proteins and immune responses, pp.
Stadecker, M.J. (1992). The role of T-cell anergy in the immunomodulation of
schistosomiasis. Parasitology Today 8, 199-204.
Stadnyk, A.W. and Gauldie, J. (1992). The acute phase protein response during
parasitic infection. Immunoparasitology Today (Immunology Today) 12
Parasitology Today 7, A7-All.
Stead, R.H. (1992). The gastrointestinal tract. Regional Immunology 4, 91-99.
Stefano, G.B., Leung, M.K., Zhao, X. and Scharrer, B. (1989a). Evidence for the
involvement of opioid neuropeptides in the adherence and migration of immuno-
competent invertebrate hemocytes. Proceedings of the National Academy of
Sciences of the USA 86, 62-30.
Stefano, G.B., Cadet, P. and Scharrer, B. (1989b). Stimulatory effects of opioid
neuropeptides on locomotory activity and conformational changes in inverte-
brate and human immunocytes: evidence for a subtype of delta and delta
receptor. Proceedings of the National Academy of Sciences of the USA 86,
6307-63 11.
Stefano, G.B., Cadet, P., Dokun, A. and Scharrer, B. (1990). A neuroimmuno-
regulatory-like mechanism responding to stress in the marine bivalve Mytilus
edulis. Brain, Behavior, and Immunity 4, 323-329.
Stefano, G.B., Cadet, P., Sinisterra, J. and Scharrer, B. (1991a). Comparative
aspects of the response of human and invertebrate immunocytes to stimulation
by opioid neuropeptides. In “Studies in Neuroscience. Comparative Aspects of
Neuropeptide Function” (E. Florey and G.B. Stefano, eds). pp. 329-334.
Manchester University Press, Manchester.
Stefano, G.B., Smith, D.M., Smith, E.M. and Hughes, T.K. (1991b). MSH can
deactivate both TNF stimulated and spontaneously active immunocytes. In
“Molluscan Neurobiology” (K.S. Kits, H.H. Boer and J. Joosse, eds), pp.
206-208. North-Holland Publishing, Amsterdam.
Stefano, G.B., Digenis, A., Spector, S., Leung, M.K., Bilfinger, T.V., Makman,
M.H., Scharrer, B. and Abumrad, N.N. (1993). Opiate-like substances in an
invertebrate, an opiate receptor of invertebrate and human immunocytes, and a
role in immunosuppression. Proceedings of the National Academy of Sciences of
the USA 90,11099-1 1103.
Sternberg, E.M., Hill, J.M., Chrousos, G., Kamlaris, T., Listwak, S.J., Gold, P.W.
and Wilder, R. (1989). Inflammatory mediator-induced hypothalamic pituitary-
adrenal axis activation is defective in streptococcal cell-wall arthritis-susceptible
Lewis rats. Proceedings of the National Academy of Sciences of the USA 86,
23762378.
Stirewalt, M.A. (1974). Schistosoma mansoni: cercaria to schistosomule. Advances
in Parasitology 12, 115-182.
Stoltz, D.B. (1993). The polydnavirus life cycle. In “Parasites and Pathogens of
Insects” (N.E. Beckage, S.N. Thompson and B.A. Frederici, eds), Vol. 1:
Parasites, pp. 167-187. Academic Press, New York.
Strand, M.R. and Dover, B.A. (1991). Development disruption of Pseudoplusia
includens and Heliothis virescens larvae by the calyx fluid and venom of
Microplitis demolitor. Archives of Insect Biochemistry and Physiology 18,
131-1 45.
Sun, S.C., Lindstrom, I., Boman, H.G., Faye, I. and Schmidt, D. (1990). Hemolin:
an insect-immune protein belonging to the immunoglobin superfamily. Science
250, 1729-1732.
Tanaka, T. and Vinson, S.B. (1991). Depression of prothoracic gland activity of
Heliothis virescence by venom and calyx fluids from the parasitoid, Cardiochiles
nigriceps. Journal of Insect Physiology 37, 139-144.
Tanaka, T. and Wago, H. (1990). Ultrastryctural and functional maturation of
teratocytes of Apanteles kariya. Archives of Insect Biochemistry and Physiology
* 13, 197-197.
Ter Maat, A. Lodder, J.C. and Wilbrink, M. (1983). Induction of egg-laying in
the pond snail Lymnaea stagnalis by environmental stimulation of the release
of ovulation hormone from the caudo-dorsal cells. International Journal of
Invertebrate Reproduction 6, 239-247.
Teunissen, Y. (1994). Molecular-biological studies of growth and reproduction in
the pond snail Lymnaea stagnalis. Thesis, Vrije Universiteit Amsterdam.
Thompson, S.N. ( 1990). Physiological alterations during parasitism and their
effects on host behaviour. In “Parasitism and Host Behaviour” (C.J. Barnard
and J.M. Behnke, eds), pp. 64-94. Taylor and Francis, London.
Thompson, S.N., Mejia-Scales, V. and Borchardt, D.B. (1992). Physiological
studies of snail-schistosome interactions and potential for improvement of in
vitro culture of schistosomes. In Vitro Cellular and Developmental Biology 27A,
497-5 14.
Thorndyke, M.C. (1990). Neurohormonal peptides in parasitic worms: a new
frontier in host-parasite pathophysiology. In “Progress in Comparative Endo-
crinology” (A. Epple, C.G. Scanes and M.H. Stetston, eds). Wiley-Liss, New
York.
Thorndyke, M.C. and Whitfield, P.J. ( 1987). Vasoactive intestinal polypeptide-
like immunoreactive tegumental cells in the digenean helminth Echinostoma
liei: possible role in host-parasite interaction. General and Comparative
Endocrinology 68, 202-207.
Thornhill, J.A., Jones, J.T. and Kusel, J.R. (1986). Increased oviposition and
growth in immature Biomphalaria glabrata after exposure to Schistosoma
, mansoni. Parasitology 93, 443-450.
Tielens, A.G.M., Van den Heuvel, J.M. and Van Eden, W. (1993). Schistosoma
mansoni: an HSP60 homologue is constitutely expressed. Experimental
Tracey, K.J. and Cerami, A. (1992). Tumor necrosis factor in the malnutrition
(cachexia) of infection and cancer. American Journal of Tropical Medicine and
Hygiene 47, 2-7.
Trottein, F., Godin, C., Pierce, R.J., Sellin, B., Taylor, M.G., Gorillot, I., Silva,
M.S., Lecocq, J.P. and Capron, A. (1992). Inter-species variation of schistosome
28-kDa glutathione S-transferases. Molecular and Biochemical Parasitology 54,
63-72.
Van der Knaap, W.P.W. and Loker, E.S. (1990). Immune mechanisms in trematode-
snail interactions. Parasitology Today 6, 175-1 82.
Van der Knaap, W.P.W., Sminia, T., Kroese, F.G.M. and Dikkeboom, R. (1981).
Elimination of bacteria from the circulation of the pond snail Lymnaea stagnalis.
Developmental and Comparative Immunology 5, 2 1-32.
Van der Knaap, W.P.W., Sminia, T., Schutte, R. and Boemgter-Barendsen, L.H.
(1983). Cytophilic receptors for foreigness and some factors which influence
phagocytosis by invertebrate leucocytes: in vitro phagocytosis by amoebocytes
of the snail Lymnaea stagnalis. Immunology 48, 377-383.
Van der Knaap, W.P.W., Boots, A.M.H., Meuleman, E.A. and Sminia, T. (1985).
Search for shared antigens in the schistosome-snail combination Trichobilharzia
ocellata-Lymnaea stagnalis. Zeitschrift fur Parasitenkunde 71, 2 19-226.
Van der Knaap, W.P.W., Meuleman, E.A. and Sminia, T. (1987). Alterations in the
internal defence system of the pond snail Lymnaea stagnalis induced by infec-
tion with the schistosome Trichobilharzia ocellata. Parasitology Research 73,
57-65.
Vankelcom, H., Carmeliet, P., Van Damme, J., Billiau, A. and Denef, C. (1989).
Production of interleukin-6 by folliculo-stellate cells of the anterior pituitary
gland in a histotypic cell aggregate culture system. Neuroendocrinology 59,
102-106.
Vinson, S.B. (1993). Interactions between the insect endocrine system and the
immune system. In “Insect Immunity” (J.P.N. Pathak, ed.), Vol. 48, pp. 103-1 11.
Kluwer Academic Publishers, Dordrecht, Boston, London.
Vreugdenhil, E., Jackson, J. F., Bouwmeester, T., Smit, A.B., Van Minnen, J., Van
Heerikhuizen, H., Klootwijk, J. and Joosse, J. (1988). Isolation, characterization
and evolutionary aspects of a cDNA clone encoding multiple neuropeptides
involved in the stereotyped egg-laying behavior of the freshwater snail
Lymnaea stagnalis. Journal of Neuroscience 8, 4 1 8 4 4191.
Wakelin, D. (1984). “Immunity to Parasites. How Animals Control Parasite
Infections.” Edward Arnold (Publishers) Ltd., London.
Wakelin, D. (1988). Genetic control of immunity to helminth infections. In
“Parasitology in Focus. Facts and Trends” (H. Mehlhorn, ed.), pp. 651-670.
Springer-Verlag, Berlin, Heidelberg.
Wani, M., Yagi, S. and Tanaka, T. (1990). Synergistic effect of venom, calyx and
teratocytes of Apanteles kariyai on the inhibition of larval pupal ecdysis of
the host, Pseudaletia separata. Entomologia Experimentalis et Applicata 57,
10 1-104.
Weigent, D.A., Cam, D.J.J. and Blalock, J.E. (1990). Bidirectional communication
between the neuroendocrine and immuno systems. Annals of the New York
Academy of Sciences 659, 17-27.
Welch, W.J. (1993). How cells respond to stress. Scientific American 268(5),
3441.
Welch, W.J., Kang, H.S., Beckmann, R.P. and Mizzen, L.A. (1991). “Response
of Mammalian Cells to Metabolic Stress; Changes in Cell Physiology and
Structure/Function of Stress Proteins.” Springer-Verlag, Berlin, Heidelberg.
Wijdenes, J., Van Elk, R. and Joosse, J. (1983). Effects of two gonadotropic
hormones on polysaccharide synthesis in the albumen gland of Lymnaea stag-
nalis, studied with the organ culture technique. General and Comparative
Endocrinology 51, 263-27 1.
Williams, C.L. and Gilbertson, D.E. (1983). Altered feeding behaviour of Schisto-
soma mansoni-infected Biomphalaria glabrata. Journal of Parasitology 69,
671-676.
Williams, D.L., Pierce, R.J., Cookson, E. and Capron, A. (1991). Molecular
cloning and sequencing of glutathione peroxidase from Schistosoma mansoni.

Molecular and Biochemical Parasitology 52, 127-1 30.
Winfield, J.B. and Jarjour, W,N. (1991). Stress proteins, autoimmunity, and auto-
immune disease. In “Current Topics in Microbiology and Immunology”. Vol.
167: Heat shock proteins and immune responses (S.H.E. Kaufmann, ed.), pp.
Yagi, Sand Tanaka, T.N. (1992). Retardation of testis development in the army-
worm, Pseudaletia separata, parasitized by the braconid wasp, Cotesia kariyai.
Invertebrate Reproduction and Development 22, 15 1-157.
Yoshino, T.P. and Lodes, M.J. (1988). Secretory protein biosynthesis in snail
hemocytes: in vitro modulation by larval schistosome excretory-secretory
products. Journal of Parasitology 74, 538-547.
Yoshino, T.P., Lodes, M.J., Rege, A.A. and Chappell, C.L. (1993). Proteinase
activity in miracidia, transformation excretory-secretory products, and primary
‘
sporocysts of Schistosoma mansoni. Journal of Parasitology 79, 23-3 1 .
Young, D.B., Mehlert, A. and Smith, D.F. (1990). Stress proteins and infectious
diseases. In “Stress Proteins in Biology and Medicine” (R. Morimoto,
A. Tessieres and C. Georgopoulos, eds), pp. 131465. Cold Spring Harbor
Laboratory Press.
Yuckenberg, P.D., Poupin, F. and Mansour, T.E. (1987). Schistosoma mansoni:
protein composition and synthesis during early development: evidence for early
synthesis of heat shock proteins. Experimental Parasitology 63, 301-3 1 1 .
Zitnan, D., Kingan, T.G., Kramer, S.J. and Beckage, N.E. (1994). Accumulation of
neuropeptides in the cerebral neurosecretory system of Manduca sexta larvae
parasitized by the braconid wasp Contesia congregata. Developmental Biology.
Submitted.
Myiasis of Humans and Domestic Animals
Martin Hall
Department of Entomology. The Natural History Museum. London. UK

and
Richard Wa II
School of Biological Sciences. University of Bristol. UK
.
1 Introduction ............................... 258
1.1. What is myiasis? . . . . . . . . . . . . . . . . . . . . . . . . . . 258
......................
1.2. Classification of myiases 259
.
2 Principal Myiasis Species and Their Life Cycles ............. 262
2.1. Oestridae .............................. 262
2.2. Calliphoridae and Sarcophagidae . . . . . . . . . . . . . . . . . . 266
. ..............
3 Evolution of Myiasis as a Life History Strategy 269
3.1. Oestridae .............................. 269
..................
3.2. Calliphoridae and Sarcophagidae 271
.
4 Current Status of Species: Their Distribution. Economic Importance and
............
Current Research on Their Behaviour and Ecology 273
4.1. Oestridae .............................. 273
4.2. Calliphoridae . . . . . . . . . . . . . . . . . . . . . . . . . . . . 279
4.3. Sarcophagidae ........................... 286
.
5 Physiology of Myiasis . . . . . . . . . . . . . . . . . . . . . . . . . . 287
5.1. Predisposing conditions for myiasis . . . . . . . . . . . . . . . . . 287
5.2. Pathology and immunology . . . . . . . . . . . . . . . . . . . . . 289
6. New and Improved Control Techniques . . . . . . . . . . . . . . . . . 293
6.1 Insecticides ............................. 293
....................
6.2. Mechanical means of control 298
..........................
6.3. Biological control 299
.......................
6.4. Sterile insect technique 299
6.5. Genetic control . . . . . . . . . . . . . . . . . . . . . . . . . . . 301
ADVANCES M PARASITOLOGY VOL 35 Copyright (D 1995 Acodemic Press Limited

ISBN 0-12-0317354 All rights of reproduction in any form resewed
258 MARTIN HALL AND RICHARD WALL
6.6. Vaccines .............................. 303

6.7. Baits, traps and targets . . . . . . . . . . . . . . . . . . . . . . . 305
7. New Monitoring, Modelling and Forecasting Methods . . . . . . . . . . 306
7.1. Monitoring by serodiagnosis .................... 306
7.2. Monitoring by traps . . . . . . . . . . . . . . . . . . . . . . . . . 307
7.3. Modelling and forecasting ..................... 308
8. Conclusions and Future . . . . . . . . . . . . . . . . . . . . . . . . . 309
Acknowledgements ........................... 31 1
References ............................... 31 1
1. INTRODUCTION
1.1. What is Myiasis?
The associations between animals and fly larvae can be categorized in a

matrix with the outcome of the association on one axis and the state of the
animal tissue on the other axis (Table 1). The subject of this review is
myiasis, where fly larvae use living hosts as a food source for growth and
development.
This paper is not intended to be a fully comprehensive historical review
of the biology of myiasis and all its associated facets. Rather, the aim is to
provide an overview of the principal recent developments in this subject.
Even in this aim, given its scope and the necessary limitation on length,
many issues have been given what may appear a relatively superficial
treatment. This is particularly so where they have been reviewed in detail
recently elsewhere, or where they were considered to be of peripheral
importance. In these cases, we hope that this review will serve as a
stimulus and guide to further enquiry.
-Kirby and Spence (1815) first used the term scholechiasis to refer to
animal diseases caused by the larvae of insects in general. Hope (1840)
later proposed the term myiasis (Greek myia = fly) to differentiate those
diseases caused by fly larvae, and he further proposed that diseases caused
by’larvae of Lepidoptera and Coleoptera be referred to as scholeciasis and
canthariasis respectively. Myiasis has since been defined as, “the infesta-
tion of live human and vertebrate animals with dipterous larvae, which, at
least for a certain period, feed on the host’s dead or living tissue, liquid
body substances, or ingested food” (Zumpt, 1965). The vertebrates usually
involved are mammals, birds, amphibians and reptiles, but a case of
aquarium fish (Asryanax mexicanus fasciatus, Mexican tetra) infested in
the stomach by Calliphora has also been reported (Bristow er al., 1990).
Myiasis is a large field with publications in many widely dispersed areas
of the literature. It is hoped that this review will bring some of these areas
MYlASlS OF HUMANS AND DOMESTIC ANIMALS 259
together and stimulate a cross-fertilization of ideas between workers in

different fields. Many references are to single case reports, particularly of
human infestations, but these will not be included here except where they
make an important, novel contribution.
Myiasis has had and still has a much greater economic impact as an
infestation of animals than of humans; there are no species of Diptera
which are restricted to humans for their development but there are many
that have a very restricted host range on animals. However, human myiasis
can be common in parts of the tropics, for example Singh et al. (1993)
reported 254 unidentified cases of myiasis over a six-year period in the Ear,
Nose and Throat Department of a single hospital in Rohtak, India. We will
concentrate on the principal myiasis agents of humans and their livestock.
We will not deal with the many species that naturally parasitize wildlife
and only rarely parasitize humans and their livestock, except where studies
of these provide important information of relevance to the myiasis condi-
tion in general. Zumpt (1965) deals with myiasis in wildlife in the Old
World, with more recent works for southern Africa being reviewed by
Horak (1987). Baumgartner (1988) provides a more recent review of
calliphorid and sarcophagid myiasis of Nearctic wildlife.
1.2. Classification of Myiases
The different forms of myiasis have been classified in two ways. First, in
anatomical terms, based on the part of the host’s body that is infested
(Bishopp, in Patton, 1922) and second, in parasitological terms, according
to the types of host-parasite relationship (Patton, 1922). The first classifi-
cation can provide a convenient short cut to identification of the fly species
concerned for practical diagnosis (Table 2), but the second gives a better
understanding of the biology of the fly as a guide to treatment or prevention
as well as providing information on the evolution of the habit.
Adopting the parasitological classification, we will consider three main
groups of myiasis-producing species: obligatory parasites, which must
Table 1 Matrix categorizing the associations between fly larvae and animals
with respect to the state of the animal host (living or dead) and the nature of the
association (detrimental or beneficial).
Detrimental Beneficial
Living hosts Myiasis (see this review) Maggot therapy (e.g. Sherman
and Pechter, 1988)
Dead hosts Food spoilage (e.g. Esser, Nutrient recycling and forensic
1991) (e.g. Smith, 1986)
Table 2 Classification of myiases according to their anatomical location in or on

the host animal. The division of myiases into five groups is based on the groupings
of Zumpt (1965) in the first column. The second and third columns show the
comparable groupings of Bishopp (in Patton, 1922) and the modification of these
by James ( 1947).
Zumpt Bishopp James
1. Sanguinivorous 1. Bloodsucking 1. Bloodsucking
2. Dermal/subdermal 2. Tissue destroying 2. Furuncular
3. Subdermal migratory 3. Creeping
4. Traumatic/wound
5. Anal/vaginal
-3. Nasopharyngeal 4. Infestations of the head 6. Nose, mouth and sinuses
passages 7. Aural
8. Ocular
4. Intestinal 5. Intestinal/urinogenital 9. Enteric
5. Anal/vaginal
5. Urinogenital 5. IntestinaVurinogenital 10. Bladder and urinary
passages
5. Anal/vaginal
develop on live hosts and facultative parasites, which can develop on both
living and dead organic matter and can be divided into two groups, the
primary species which are able to initiate myiasis and the secondary
species which occur after obligate or primary species have initiated it
(Zumpt, 1965).
A fourth grouping, the accidental myiases or pseudomyiases (Zumpt,
1965), occur when fly eggs or larvae are inadvertently swallowed. In a
cgntrolled experiment, nausea, vomiting, intestinal cramps and diarrhoea
were observed in volunteers who swallowed live larvae of Musca domes-
tics, Calliphora or Sarcophaga in gelatine capsules (Kenney, 1945).
Naturally acquired accidental intestinal infestations can show similar
symptoms or be benign (Nagakura et al., 1991).
Miscellaneous myiases are often reported, particularly in humans, that
are essentially accidental but do not involve the intestinal tract. They occur
when the wrong host is invaded or eggs are laid in atypical sites for the
species. Examples are facultative species involved in human urinary tract
myiases, when eggs are deposited at the entrance to the urethra, hatch into
larvae and develop at the end of the urethra (Musca, Gupta et al., 1982) or
inside the urethra or bladder, being passed in the urine (Megaselia, Singh
and Ranah, 1989; Piophilu, Saleh and El Sibae, 1993). Another internal
region of the human body that can be unusually infested is the respiratory
tract, with relatively mild (Megaselia, Carpenter and Chastain, 1992) or
MVlASlS OF HUMANS AND DOMESTIC ANIMALS 261
severe (Megaselia, Komari ef al., 1978) symptoms. Such myiases are

relatively uncommon but, as they involve humans, tend to be reported in
the literature with a frequency that exaggerates their importance. Hall and
Smith (1993) review such cases and so they will not be considered further
Table 3 Table of relationships between the taxonomic groupings of the myiasis

agents of humans and domesticated animals, their degree of parasitism and their
main anatomical site of parasitism (facultative 1 = primary; facultative 2 =
secondary).
~~
Taxonomic grouping Degree of parasitism Site of parasitism

OESTRIDAE
Oestrinae Obligate Nasopharyngeal
Oestrus
Rhinoestrus
Cephenemyia
Cephalopina
Gasterophilinae Obligate Digestive tract
Gasterophilus
Hypodermatinae Obligate Dermal, furuncular
Hypoderma
Przhevalskiana
Cuterebrinae Obligate Dermal, furuncular
Cuterebra
Dermatobia
CALLIPHORIDAE and SARCOPHAGIDAE
Cochliomyia hominivorax Obligate Dermal, wound
Chrysomya bezziana Obligate Dermal, wound
Wohlfahrtia magnifrca Obligate Dermal, wound
Wohlfahrtia vigil Obligate Dermal, furuncular
Auchmeromyia Obligate Sanguinivorous
Cordylobia Obligate Dermal, furuncular
Lucilia sericata Facultative 1 Dermal, wound
Lucilia cuprina Facultative 1 Dermal, wound
Other Calliphoridae and Sarcophagidae species in genera:
Cochliomyia Facultative 1 and 2 Dermal, wound
Chrysomya Facultative 1 and 2 Dermal, wound
Lucilia Facultative 1 and 2 Dermal, wound
Calliphora Facultative 1 and 2 Dermal, wound
Phormia Facultative 1 and 2 Dermal, wound
Protophormia Facultative 1 and 2 Dermal, wound
Sarcophaga Facultative 1 and 2 Dermal, wound
Wohlfahrtia Facultative 1 and 2 Dermal, wound
here, except where they involve species important in their true host, for
example, Oestrus causing ophthalmomyiasis and Gasterophilus causing so-
called creeping myiasis in the dermal layers of humans. Chodosh and
Clarridge (1992) review ophthalmomyiasis of humans.
The three major families of myiasis-producing flies are the Oestridae, as
recognized by Wood (1987) (four subfamilies, Oestrinae, Gasterophilinae,
Hypodermatinae and Cuterebrinae), the Calliphoridae (blowflies) and the
Sarcophagidae (fleshflies) and this review will concentrate on them. They
all belong to the superfamily Oestroidae (Calyptrate) (McAlpine, 1989).
Their involvement in myiasis is tabulated from an entomological perspec-
tive, but taking account of the degree of parasitism and anatomical loca-
tion, in Table 3. Works containing detailed taxonomic identification keys to
'myiasis species are James (1947), Zumpt (1963, Guimariies et al. (1983),
Spradbery (1991) and Hall and Smith (1993).
2. PRINCIPAL MYlASlS SPECIES AND THEIR LIFE CYCLES
The Oestridae (ca 151 species in 28 genera; Wood, 1987) are all obligate
parasites and all of the nutritional requirements for adulthood are taken
from their hosts during the larval stage. Adults have atrophied mouthparts
and do not feed, although Cephenemyia and Cuterebra may imbibe fluids
(Wood, 1987).
In contrast to the Oestridae, the Calliphoridae (ca 1000+ species in 150
genera; Shewell, 1987a) and Sarcophagidae (ca 2000+ species in 400
genera; Shewell, 1987b) include both obligate and facultative parasites
and all feed both as larvae and as adults. At least 80 species in these two
families have been recorded as agents of myiasis (Zumpt, 1965). However,
only a relatively small number of species, three obligate screwworms
(Cochliomyia hominivorax, Chrysomya bezziana and Wohlfahrtia magni-
&a) and two species of primary facultative blowflies (Lucilia sericata and
Luqilia cuprina) are of major clinical and economic importance worldwide.
They will, therefore, be the focus of future discussion.
2.1. Oestridae
2.1.1, Oestrinae
The genus Oestrus includes 0. ovis, the sheep nasal bot fly, whose larvae
develop in the head sinuses and nasal passages of sheep and goats in all
sheep-farming areas of the world. Females can produce up to 500 larvae
which are deposited directly into the hosts’ nostrils (Kettle, 1990). Adult
fly activity can cause panic, sheep burying their heads in other sheep or
running about snorting in an effort to dislodge larvae. Oestrus ovis larvae
are well adapted to their location in the nasal passages of sheep and goats,
with a variation in spinulation between instars that accords with the
requirements of the instar for anchoring within the respiratory tract
(Guitton and Dorchies, 1993). Effects of infestation can be insignificant
or severe (especially in lambs), with purulent discharge from nostrils,
repeated sneezing and shaking of head and breathing difficulties. Rarely,
larvae may enter the brain causing a condition known as “false gid”
(ataxia, circling and head pressing). Development takes 25-35 days in
summer but is extended through the winter when first instars may last up
to 9 months, giving a total development time of 10-11 months.
Larvae of the Old World genus Rhinoestrus, infest the nasal sinuses of
their hosts and are, generally, very host specific: R . purpureus attacks
horses and donkeys. The genus Gedoelstia generally parasitizes ante-
lopes, but can cause a’serious myiasis of sheep in the western parts of
southern Africa (Zumpt, 1965).
Cephalopina titillator, the camel nasal bot fly, is the only species in its
genus and the larvae develop in the nasal cavities of camels wherever
camels naturally occur, even in areas of introduction such as Australia
(Spratt, 1984). High levels of infestation may occur in camel herds, up to
91% in the rainy season in Nigeria (Desbordes and Ajogi, 1993), and the
presence of larvae in their nostrils may cause considerable irritation,
difficulty in breathing and, exceptionally, death following complications
due to secondary infections (Higgins, 1985).
Important throat bot flies of reindeer are Cephenemyia trompe (reindeer
throat bot fly) and C. auribarbis (red deer throat bot fly).
Oestrus ovis and Rhinoestrus purpureus can cause ophthalrnomyiasis in
humans. Other Oestrinae reported to cause human ophthalmomyiasis
include Pharyngomia picta, the deer throat bot fly (Sauter and Huber, 1988).
2.1.2. Gasterophilinae
Originally restricted to the Palaearctic and Afrotropical regions, species of
Gasterophilus (bot flies) now have a worldwide distribution. Their larvae
develop in the digestive tract of equids. Eight species of Gasterophifus are
generally recognized and all but one of those with known biology lay their
eggs directly on the host, attaching them to the hairs in particular body
regions such as cheek, chin, lip, leg and mane (Cogley, 1991). The numbers
of eggs produced depends on the species, ranging from about 160 for G .
haemorrhoidalis to over 2000 for G . pecorum (Kettle, 1990). A combina-
tion of morphological specialization of the eggshell (attachment organ) and
an adhesive ensures effective long-term egg attachment (Cogley and

Anderson, 1983). Published observations of adult fly behaviour are rare,
but Cope and Catfs (1991) have published an account of the behaviour of
G. intestinalis around horses in the USA. This demonstrates the remarkable
tenacity of ovipositing females which will lay eggs on walking and trotting
horses in addition to standing horses. If this action induces the horse to
gallop, the females will pursue galloping horses alongside the flank until
the host stops when the flies will immediately resume oviposition. This
urgency to oviposit reflects the short life span of the adults, mature
ovipositing females have an effective field life span of one day (Cope
and Catts, 1991). A brief outline :of the biologies of the six important
species is provided in Table 4.
2.1.3. Hypodermatinae
Hypoderma are the heel flies, warble flies or cattle grubs whose larvae
develop in subcutaneous “warbles” which spoil the hosts’ hides and cause
serious losses to the cattle industry of the Holarctic region (Scholl, 1993).
The important species are H . bovis and H . lineatum. Mating takes place off
the host at aggregation points where females are intercepted in flight.
Females are reproductively well adapted to their short, non-feeding life
style, because they emerge from the puparium with all their eggs fully
developed, and the capacity to mate immediately and oviposit on host
Table 4 Outline of the biologies of important species of Gasterophilus.

Gasterophilus nigrocornis: eggs laid on cheeks, hatch and larvae penetrate into
cheeks; once second instar they are swallowed and embed themselves in duodenum.
Gasterophilus haemorrhoidalis: eggs attached to hairs fringing lips, moisture from
licking is needed for incubation; larvae penetrate epidermis and move to stomach
and duodenum; third instars move to rectum and attach very close to anus.
Gasterophilus inermis: eggs laid on cheeks, larvae migrate in epidermis to mouth
(hairs fall out along its path); second and third instar larvae in rectum.
’
Gasterophilus intestinalis: eggs laid mainly on inner forelegs and stimulated to
emerge by licking; larvae migrate through tongue (for 24-28 days); second instars
move to stomach and cluster at boundary of glandular and non-glandular
epithelium.
Gasterophilus nasalis: eggs laid underneath jaw and hatch in 5-10 days; larvae
migrate to lips and develop in pus-pockets of necrosis in gums to second instar,
then move to duodenum; passed with faeces about 11 months after hatching.
Gasterophilus pecorum: larvae laid in grass where they remain viable for months,
until ingested by horse; they burrow into mucous membranes and may remain at
the root of the tongue for 9-10 months, or be swallowed and settle in the pharynx,
oesophagus or stomach.
cattle (Scholl and Weintraub, 1988). Eggs are laid on the host’s hairs,
either singly ( H . bovis) or in batches of up to 15 ( H . lineatum) on the
lower region of the legs and lower body. The persistence of the females in
laying 300-800 eggs can cause a dramatic escape response, “gadding”, by
the hosts. The most obvious economic effects are due to the creation of
holes in hides once the larvae have reached the back after their migration
from sites of oviposition via the spinal column ( H . bovis) or oesophagus
( H . lineatum). Losses due to decreased weight gain and milk production
may be more significant in economic terms (Steelman, 1976; Tarry, 1986).
In a similar manner to the cattle pests, Hypoderma diana parasitizes deer
aqd H . tarandi (= Oedemagena tarandi, Wood, 1987) parasitizes reindeer.
A related warble fly is the goat warble of the Mediterranean Basin,
Przhevalskiana silenus. Rarely, the larvae of Strobiloestrus species, para-
sites of antelopes in the family Bovidae (e.g. klipspringers, mountain
reedbuck, steenbok), cause a furuncular myiasis of cattle (Horak and
Boomker, 1981) and sheep (Brain et al., 1983).
2.1.4. Cuterebrinae
The most important Cuterebrid is Dermatobia hominis (tbrsalo, beme,
human bot fly), a Central and South American species whose larvae create
boil-like swellings where they enter the skin (Lane et al., 1987). In
economic terms it is most important as a pest of cattle, but it also para-
sitizes man, dogs and a variety of other domestic and wild animals and
birds. The hides of infested cattle can be made worthless. Dermatobia
hominis do not feed as adults and live for only a short time (ca. 6 days).
A noteworthy adaptation to this appears to be the early degeneration of
less-developed cytoblasts in the ovaries of pupae, cytoblasts that would not
have time to complete their oogenesis in the adult (Secco et al., 1992).
Dermatobia have a fascinating method of infecting their hosts. Whereas
other Cuterebridae lay eggs on substrate near or within the entrance to host
nests (Beamer et al., 1943; Catts, 1967; Capelle, 1970) or on grass near
trails used by hosts (Beamer, 1950), D . hominis lay their eggs onto host-
visiting, carrier (phoretic) insects. The eggs incubate on the carrier and
then, when the carrier next visits a host, first instar larvae hatch in response
to the sudden temperature rise near the host’s body, leaving the egg through
a well-developed, plate-like operculum at the anterior end (Mourier and
Banegas, 1970). The larvae invade the host’s subcutaneous tissues where
they remain for 35-42 days. Although eggs are ready to hatch after 4 days
at 30”C, larvae may emerge from the same batch over a considerable period
(e.g. from 5-20 days at 28°C; Mourier and Banegas, 1970), which might
help to improve dispersion of larvae.
In the laboratory, females held in conditions of constant dark laid only
5% of the number of eggs laid by females in constant light, therefore

Mourier and Banegas (1970) concluded that most egg-laying in nature
occurred during the day. This is supported by the fact that all phoretic
vectors recorded have been day-flying species. There are few observations
of where Dermatobia catch their vectors, but Mourier and Banegas (1970)
concluded that it must be on or near cattle. Other hosts may also serve as
catching sites (M.J.R. Hall, unpublished observation, on horse in Bolivia).
Females have the potential to lay up to 1000 eggs (Catts, 1982). The eggs
are laid side by side along their long axis, usually on one side of the
abdomen of the carrier insect. Rate of egg-laying is about one per second
and more eggs are laid on heavier vectors, with a mean of 28 per vector
(Sarcopromusca sp.). Females usually oviposit only on live flies. Inanimate
objects, including dead flies, are rejected (Mourier and Banegas, 1970).
Excellent sources of information for all aspects of work on D. hominis
are provided by the bibliographies of Guimariies and Papavero (1 966) and
Guimariies et al. (1983).
Cuterebra species cause myiasis of rodents, lagomorphs and, occasion-
ally, humans in North America (Baird et al., 1989).
2.2. Calliphoridae and Sarcophagidae
The calliphorid and sarcophagid agents of myiasis have broadly similar life
cycles except for Auchmeromyia and Cordylobia. The latter two genera are
discussed separately at the end of this more general discussion about the
remaining genera.
For most genera, eggs or first instar larvae are laid on or in the host. The
larvae pass through three instars while feeding on the host tissues, before
entering a wandering stage in which they migrate away from the strike
focus and, after a period of dispersal, pupate, prior to adult emergence.
The New World screwworm fly, Cochliomyia hominivorax, is an obligate
ectoparasite and will infest almost all warm-blooded livestock, wildlife and
humans; it is unable to breed in carrion (James, 1947). Females are auto-
genous, mate during early vitellogenesis and oviposit approximately every
three days (Guillot et al., 1977). They lay batches of up to 500 eggs, with a
mean of 200 per batch, at the edges of open wounds or in body orifices
(Thomas and Mangan, 1989). Within 24 h of hatching, the maggots start to
feed, burrowing into the living tissue. The behavioural, ecological and
physiological characteristics of the different phases (adolescent, sexual
and reproductive) in adult screwworm life, are reviewed by Thomas
(1993a). The extensive wound which results from larval feeding may
rapidly lead to the death of the struck animal.
The Old World screwworm fly Chrysomya bezziana is similar in many
respects to Cochliomyia hominivorux, being an obligate parasite of

mammals and unable to develop in carrion (Zumpt, 1965). Females are
normally autogenous (Spradbery and Sands, 1981) and lay batches of 175
eggs on average, at the edges of wounds or body orifices. The eggs hatch in
10-12 h at 37°C and the first instar larvae migrate to the wound or moist
tissue where they aggregate and begin feeding on the wound fluids. The
mean female life expectancy is about 9 days, resulting in a mean lifetime
fecundity for C. bezziana of about 146 female progeny (Spradbery and
Vogt, 1993). The feeding activity of the larvae may cause serious tissue
damage, resulting in loss of condition, injury to the hide, secondary
invasion and often death (Humphrey et al., 1980). There is no evidence
to support the assertion that C. bezziana is a less aggressive species than
C. hominivorax or that the effects of its myiasis are less pathogenic
(Spradbery, 1994).
Both Lucilia sericata and L. cuprina are primarily anautogenous. The
maximum number of eggs that can be matured by L. cuprina is dependent
on size (Vogt et al., 1985a) and protein availability; when protein is freely
available an average of about 250 eggs are produced in each batch (Barton
Browne et al., 1979). However, under field conditions in Australia, female
L. cuprina rarely obtain sufficient proteinaceous material to mature their
full egg complement (Vogt et al., 1985a). For L. sericata, egg production is
also dependent on size and protein availability, but protein does not appear
to be limiting in the field under British conditions and females produce an
average of about 225 eggs per batch (Wall, 1993). Mean life expectancy,
estimated from field age distributions, is temperature dependent and has
been estimated to be between 3 and 6 days at average ambient temperatures
of 19°C and 16"C, respectively. At these mortality rates, about 15% of
females would be expected to reach a first oviposition, 7% to reach a
second and only 3% to reach a third, giving an expected mean lifetime
reproductive output of about 50 eggs per female (Wall, 1993).
The sarcophagid, Wohlfahrt's wound myiasis fly, Wohljiahrtia magnifica
is an obligate, larval parasite of warm-blooded vertebrates throughout the
Mediterranean basin, east and central Europe and Asia Minor. Due to the
larviposition habit, it can cause a rapid and severe myiasis in humans and
most livestock, including poultry (Zumpt, 1965). Female flies deposit
120-170 first instar larvae at sites of wounding on the host, or beside its
body orifices. The larvae feed and mature in 5-7 days and then leave the
wound for pupation (Zumpt, 1965; Kettle, 1990). The major work on the
biology of the species is still that of Portschinsky (1916).
Wohlfahrtia nuba also infests wounds of livestock in North Africa and
the Middle East, but it probably feeds only on dead or diseased tissues
rather than on living tissues (James, 1947).
Whereas W. magnifica causes a traumatic cutaneous myiasis, two species
of Wohvahrtia cause a furuncular myiasis in North America. Larvae of W .

vigil can penetrate intact skin if it is thin and tender, hence it is the young
of animals and humans that are most affected. The larvae produce furuncles
similar to those of Dermatobia but containing up to five larvae (O’Rourke,
1954; Alexander, 1984). In Europe W . meigeni has not been recorded as a
myiasis agent, but it has been a serious pest to the mink and fox-farming
industry in North America (Gassner and James, 1948: as W . opaca) and can
also cause a myiasis in infants similar to that caused by W . vigil (Haufe and
Nelson, 1957).
Myiasis due to Sarcophaga species is of no significance in veterinary
terms: many species breed in excrement, carrion and other decomposing
organic matter and are only occasionally involved in myiasis. They are
rarely involved in myiasis of humans, either in wound myiasis (Arbit et al.,
1986) or in intestinal myiasis due to accidental ingestion (Hasegawa et al.,
1992). Sarcophaga cruentata (= haernorrhoidalis) is one of the most
common species and breeds mainly in faeces (Zumpt, 1965; Smith, 1986).
2.2.1. Auchmeromyia
Bloodsucking larvae of the African species Auchmeromyia senegalensis
the Congo floor maggot, are atypical myiasis species as they do not live on
or in the host, but are obligate, haematophagous ectoparasites and suck the
blood of sleeping humans (Noireau, 1992) and burrow-dwelling animals
(sanguinivorous myiasis). In this respect their behaviour is more like that
of an adult, bloodsucking insect rather than a myiasis species, to the extent
that they have been shown experimentally to be capable of transmitting
trypanosomes (Geigy and Kauffmann, 1977). Somewhat similar behaviour
is shown by blood-feeding blowfly larvae of the genus Protocalliphora.
These are obligate parasites of nestling birds (Sabrosky et al., 1989;
Bennett and Whitworth, 1991) but, in general, their effects are not serious
(Whitworth and Bennett, 1992; Johnson and Albrecht, 1993).
2.2.2. Cordylobia
Cordylobia includes C . anthropophaga, the “tumbu” fly of Africa, which
causes a boil-like (furuncular) type of myiasis like that of D . hominis in the
Americas. Its biology has been investigated most extensively by Blacklock
and Thompson (1923), who concluded that wild rats were the most impor-
tant natural reservoir of infection in Sierra Leone. However, humans and
dogs are the most important hosts in economic terms. Eggs are not
deposited directly onto a host, rather onto dry, shaded ground, especially
if contaminated by urine and faeces. They may also be laid on drying
laundry. Larvae hatch in 1-3 days and remain just under the soil surface
until activated by host body heat. They then emerge, burrow into the host
and grow for 8-15 days in a furuncle, usually with only one larva per
abscess. Cordylobia rodhaini only rarely infests man (Kremer ef al., 1970).
3. EVOLUTION OF MYlASlS AS A LIFE HISTORY STRATEGY
This section will concentrate on the evolution of the myiasis habit itself
rather than the evolution of the groups involved in myiasis, although the
two are interlinked. The phylogeny of the three principal myiasis families,
Oestridae, Calliphoridae and Sarcophagidae, has been considered in detail
by McAlpine (1989) and Pape (1992).
The family and subfamily groups can be characterized with respect to their
feeding habits (Schaefer, 1979). Most Calliphoridae and Sarcophagidae are
catholic in the range of hosts they exploit and their selection of hosts is
frequently a reflection of host availability. Thus, in much of the Americas,
Cochliomyia hominivorax is principally a pest of cattle but in Libya it was
mainly found on sheep, the most numerous local hosts (FAO, 1992). In
contrast, most Oestridae show a high degree of host specificity. The
specialization of some Oestridae on endangered hosts could, of course,
lead to their extinction, for example, Gyrostigma on rhinoceroses (Cogley,
1990).
Zumpt (1965) postulated two roots for the evolution of myiasis: (1) the
sanguinivorous root, with the specialized oestrid behaviours derived from
less-specialized calliphorids that had ectoparasitic bloodsucking larvae,
like the present day Auchmeromyia (on mammals) and Protocalliphora
(on birds), and, (2) the saprophagous root, with an evolutionary trend from
carrion breeders to obligate wound myiasis species, as outlined below.
James (1969) noted that the transition to parasitism, especially faculta-
tive, involved a broad adaptability or lack of specialization of a group in a
state of biological flux, of relatively youthful and vigorous stock.
3.1. Oestridae
Papavero (1977) speculates on the evolution of Oestrinae (as Oestridae) in

relationship to their hosts, 25 genera in four orders (Marsupialia, Probo-
scidea, Artiodactyla and Perissodactyla). The hosts have the following
characteristics: (1) terrestrial herbivores; (2) minimum length (1 m) and
weight (20-25 kg), (3) in the geographical and ecological area of the
parasite for sufficient time to allow parasite-host adaptation; (4) gregarious
if inhabiting open situations.
In his discussion, Schaefer (1979) included the other oestrid subfamilies

in addition to Oestrinae. He observed that their hosts either lived in herds
(because animals in-herds are more easy to find than single animals,
although this depends on their distribution), or in burrows (because a
burrow which does not move and can accumulate attractive odours over
many years is more easy to find than a single animal). The only oestrid that
naturally parasitizes carnivores is Dermatobia hominis, although it mainly
parasitizes herbivores. Why is it that oestrid hosts are mainly herbivorous?
The major reason is probably that herbivores are more numerous as hosts
than carnivores, and so supply more habitat (Schaefer, 1979). For example,
on Isle Royale in Lake Superior, USA, the ratio of the numbers of moose to
. their only predator, wolves, was estimated to be 42: 1 when the populations
were relatively stable, giving a prey-predator biomass ratio of 452: 1
(Jordan et al., 1971). Likewise, prey-predator biomass ratios in five
African game reserves ranged from 94:l to 301:l (predators were lion,
leopard, cheetah, spotted hyena and wild dog: Schaller, 1972). For the
Gasterophilinae, an additional reason might be that the digestive tract of
herbivores is a less hazardous environment for larvae than the digestive
tract of carnivores.
The evolution of the ovipositional behaviour of Dermatobia hominis
deserves special attention because it is unique. Today there is a heavy
infestation of domestic animals with Dermatobia hominis. However, before
the introduction of cattle and other domestic animals to the Americas in the
16th century, the main hosts were native mammalian and avian fauna, e.g.
monkeys, wild pigs, jaguars, pumas, agoutis, grisons, armadillos, toucans
and ant birds. Townsend (1935) postulated that before the advent of cattle,
Dermatobia used only forest mosquitoes as egg carriers, and that the hosts
were only the native tropical American mammalian fauna which are,
generally, thinner skinned than cattle. In order to exploit the increasing
numbers of cattle as hosts, Townsend speculated that development of the
carrier habit was greatly stimulated, and that it is still developing now.
Developments were needed to overcome problems associated with the
more open and drier environment preferred by cattle to the original forest
habitats of the fly. Townsend (1935) argued that sunlight is harmful to the
developing larva in its egg and that there was, therefore, a gradation in
acceptability of oviposition sites, with hairs on the host being worst,
phoretic flies the best and path-side vegetation in between. Humidities of
>70% are favourable for egg hatching (Mourier and Banegas, 1970). As
eggs require 4-9 days to incubate, if they were located directly on the
host’s body they would be exposed to adverse climatic conditions in the
tropics. In temperate climates, the exposure to adverse conditions on the
host is not such a problem and so there has not been the same selective
pressure on species such as Hypoderma to deposit their eggs elsewhere.
MYlASlS OF HUMANS AND DOMESTIC ANIMALS 27 1
The eggs of Gasterophilus species appear to be more resistant to adverse

tropical conditions than those of Dermatobia. At the extreme, developed
first instar lapae of G . pecorum remain viable inside egg shells for many
months (Zumpt, 1965).
Mourier and Banegas (1970) speculate that the adaptive value of oviposi-
tion on phoretic carriers may be to camouflage eggs, so that the hosts do not
develop defence mechanisms. This might be more of a problem if the
original hosts were efficient groomers, such as rodents and primates, but
would seem unlikely to be important on bovid hosts, as demonstrated by the
success of hypodermatids and gasterophilids. With some Gasterophilus it is
the hosts grooming (licking) that actually stimulates hatching (Table 4).
The selection of phoretic flies by female Dermatobia hominis deserves
study from an evolutionary point of view. Clearly the best vectors of eggs
will be flies that are associated with hosts. The most likely site for
Dermatobia to come into contact with such flies is on the host, and
Mourier and Banegas (1970) believed that, in the field, female D . hominis
catch vector flies on or near cattle. Possibly the fly started out as a parasite,
laying its eggs directly on the host before, secondarily, laying on phoretic
vectors. However, this would rule out the possibility of sudden warming as
an egg-hatching stimulus, a characteristic of all cuterebrids including
Dermatobia: therefore, D . hominis is unlikely to have used egg deposition
directly on the host (Catts, 1982). The importance of the sudden rise in
temperature for hatching of cuterebrids is illustrated by studies of the
California rodent bot fly, Cuterebra latifrons (Catts, 1967). Eggs of this
species mature in about ten days at 25°C and at humidities from 11.05% to
saturated. If there are no hatching stimuli the eggs can generally remain
infective at 1525°C and 4 5 4 4 % RH for 2-3 months, even up to four
months. At 20-25"C, hatching could be stimulated by an increase in
temperature as small as 2.5"C in a period of 30 s. More gradual rises in
temperature did not stimulate hatching, e.g. 2 4 ° C over 30-60 min or 16°C
over a 4 h period. Such studies of Cuterebra are important for comparison
with Dermutobia (Table 5 ) .
Scholl and Weintraub (1988) discussed the gonotrophic development of
Hypoderma which is adapted for the short-lived, non-feeding adult stage
and suggested that a study of patterns of ovarial development among
oestrids would be of interest from an evolutionary perspective.
3.2. Calliphoridae and Sarcophagidae
The Calliphoridae and Sarcophagidae responsible for myiasis can be

divided generally into three functional groups based on their larval feeding
habits:
Table 5 Comparison of biology of the Cuterebridae genera Cuterebra and

Dermatobia based on data presented in Catts (1982).
Cuterebra Dermatobia
Relatively host specific with poor Relatively large host range and broad
tolerance of different hosts tolerance of different hosts
Rarely infest humans Frequently infest humans
Oviposition on inanimate objects near Oviposition on “porter flies”
nest or pathways of host associated with hosts
L1 develop in egg in 4-10 days L1 develop in egg in 4-9 days
Eggs hatch in response to sudden slight EggS hatch in response to sudden
. rise in temperature slight rise in temperature
L1 enter host via moist natural openings L1 enter host via bite of porter flies or
or skin lacerations directly through intact skin
Larvae undergo subcutaneous migration to Larvae do not migrate but mature near
specific maturation site to entry site virtually anywhere on
host
1. saprophages normally living in decaying organic matter and animal

carcasses, which cannot initiate a myiasis but which may secondarily
invade existing infestations (i.e. secondary facultative species);
2. facultative ectoparasites, normally adopting an ectoparasitic habit and
which are capable of initiating myiases but which occasionally live as
facultative saprophages (i.e. primary facultative species);
3. primary, obligate parasites feeding only on the tissues of living
vertebrates, usually mammals and birds.
This functional division may also reflect the evolution of the parasitic
habit. Generalized free-living saprophagous feeders, such as calliphorids,
which may occasionally act as agents of myiasis in wounded, dying or
otherwise clinically predisposed animals, may have formed the ancestral
origins of the parasitic habit. These then gave rise to facultative ectopara-
sites, attracted to skin soiled by faeces, bacterial infection and suppurating
wounds, such as Lucilia sericata or L. cuprina, which behave as primary
myiasis agents rather than saprophages. From this intermediate stage, the
obligate parasites, Cochliomyia hominivorax, Chrysomya bezziana and
Wohlfahrtia magnifrca developed (Zumpt, 1965; Erzinqlioglu, 1989).
Hence, it would appear that these three screwworm species evolved in
parallel, independently, in different parts of the world from necrophagous
ancestors, with species of the genera such as Lucilia, representing a mid
point in this development. Intermediate evolutionary stages within the
genera of the three obligate parasites themselves are represented by
Cochliomyia macellaria, Chrysomya megacephala, and Wohlfahrtia

meigeni.
If the myiasis habit of Calliphoridae and Sarcophagidae evolved from
saprophagy, then it probably involved a change in the timing of attack.
Thus, the primitive behaviour shown by carrion-breeding species changed,
through attack on stressed or dying animals, to attack on healthy animals,
i.e. from vertebrate saprophage, to feeder on vertebrate wounds, to obligate
cause of vertebrate wounds. The ectoparasites of the obligate set do not
inevitably cause death of their hosts. However, the blowfly Lucilia bufo-
nivora which is an obligate, specialist agent of myiasis of frogs and toads,
generally does cause death of its amphibian hosts and can therefore be
more correctly classified as a parasitoid (Eggleton and Belshaw, 1992).
The myiasis habit of the obligate ectoparasites may be a well-established
life history strategy in evolutionary terms. In contrast, inter-and intra-
specific variation in myiasis propensity and historical records of changes
in myiasis incidence or species of importance, provide a strong case to
suggest that some of the species responsible for facultative myiasis may
have co-evolved with the domestication and spread of livestock and the
intensification of their husbandry, and may have expanded relatively
rapidly and recently into the vacant niches so created. Possibly they were
pre-adapted as general parasites of wildlife, and so were able to take
advantage of the spread of domestic, relatively non-resistant, animals.
This may be particularly true for species of the genus Lucilia.
4. CURRENT STATUS OF SPECIES: THEIR DISTRIBUTION,

ECONOMIC IMPORTANCE AND CURRENT RESEARCH ON THEIR
BEHAVIOUR AND ECOLOGY
4.1. Oestridae
4.1.1. Oestrinae
Adult Oestrus ovis seriously annoy sheep as they deposit larvae, leading to
a loss of grazing time and condition of the sheep. Horak and Snijders
(1974) demonstrated a poorer weight gain of 0. ovis infested lambs
compared to those freed of the parasite by rafoxanide treatment, whereas
Ilchmann et al. (1986) reported losses in production ranging from 1.1 to
4.6 kg of meat, 200 to 500 g of wool and up to 10% of milk. Recent studies
have shown infestation levels in sheep of 6 5 2 % (Zimbabwe; Pandey,
1989), up to 69% (India; Jagannath et al., 1989b), up to 100% (Morocco;
Pandey and Ouhelli, 1984: South Africa; Louw, 1989: Brazil; Ribeiro et
al., 1990) and with a seasonal variation of 44-88% (France; Yilma and
Dorchies, 1991 - who discuss other recent epidemiological studies).
Yilma and Dorchies (1991) observed a maximum of three fly generations
per year in southwest France, with adults peaking in numbers in three
periods, March-April, June-July and September-October. They suggest
that treating sheep during these three periods would considerably reduce
the incidence of disease locally. In Britain the general prevalence of 0. ovis
infestation is e l % , but it can be much higher in local “hot-spots”, with
most infestations in the south of England and Wales (P. Bates, personal
communication), from where human ophthalmomyiasis has been reported
(Stevens et al., 1991).
Cases of human ophthalmomyiasis due to the larvae of 0. ovis are
frequently published, in particular from the Middle East and Mediterranean
Basin (Cameron et al., 1991; Mariotti and Vacheret, 1992; Amr et al.,
1993; Hira et al., 1993). Most cases of ophthalmomyiasis due to 0. ovis
resolve rapidly as the larvae are unable to develop beyond the first instar.
Infrequently, nasal myiasis due to 0. ovis is reported in humans (Quesada
et al., 1990).
In the Caspian region of the former USSR, Rastegayev ( 1 984) reported a
prevalence of infection of horses with Rhinoestrus of 96.7-loo%, the
major species being R . latifrons and R . purpureus. Between 45 and 899
Rhinoestrus larvae were found in the head cavities of infested horses, with
a mean of 154. Rastegayev (1984) made detailed records of the biology,
life cycle and behaviour of these species, including the observation that
female Rhinoestrus can insert larvae into the nostrils of horses not only
while in flight, but also when landed on the ground or on other objects near
the host. Thus, females can raise themselves on their tarsi and launch
several batches of larvae into the current of air inhaled by the horse.
Zayed et al. (1993) recorded the prevalence of R . purpureus in donkeys
in Egypt and showed that larvae were present from January to October with
two peaks of 100% monthly infestation, in March and July-August. No
’
infestations were found in the winter, November-December, and it was
concluded that there were two generations of the fly per year. The mean
monthly larval burden was greatest in June (58 per donkey). All first instar
and the majority (94%)of second instar R. purpureus were located in the
ethmoid bone of donkeys, whereas third instars were mostly (42%)found
in the sphenopalative communication, prior to their maturation and migra-
tion to the exterior via the common and nasal meatus and the nostrils
(Zayed and Hilali, 1993).
4.1.2. Gasterophilinae
The high value of many horses, the recurrent expense of treatments and
possible self-injury by horses under “attack” from ovipositing females
make Gasterophilus intestinalis a major economic pest of equines in North

America (Cope and Catts, 1991), despite the fact that these parasites are
frequently well tolerated. Pandey et al. (1992) review reports of Gastero-
philus in horses and donkeys which can be very high: in Morocco, 98% of
donkeys harboured G . intestinalis and 95% harboured G . nasalis. The two
species were found in almost equal proportions in donkeys but G . intesti-
nalis were more common in horses (Pandey et al. 1980). The mean
monthly burden of second and third instars of both species in donkeys
varied from 75 to 3 11, with a maximum of 715. In a major study of horses
around the Caspian Sea, Rastegayev (1984) reported 100% infection with
Gqsterophilus, with 394525 larvae per horse (mean 322). Pandey et al.
(1992) observed erosions, ulcers, nodular growths and stomach perforation
as a result of Gasterophilus infestation of donkeys. Other workers have
also incriminated Gasterophilus in gastric abscesses, ruptures, peritonitis,
general debilitation in heavy infections and even rectal prolapse (Daoud et
al., 1989). Pandey et al. (1992) conclude that treatment against bots is
justified, especially if linked with treatments for helminth parasites to
increase the performance of equines during periods of agricultural opera-
tions. On rare occasions humans can acquire cutaneous or oral infections
with Gasterophilus (Hall and Smith, 1993).
4.1.3. Hypodermatinae
The gadding behaviour of cattle irritated by ovipositing Hypoderma is
thought to be a potential cause of injury, spontaneous abortion and reduced
milk production, but these losses have not been assessed (Scholl, 1993).
Likewise, the effects of Hypoderma on weight gain are open to interpreta-
tion and Scholl (1993) points out that many questions regarding effects of
infection remain to be answered, including the threshold levels of infesta-
tion for economic loss. However, the effect of Hypoderma on hides is well
established.
A programme of Hypoderma eradication in Britain was launched in
1978, at which time annual losses in the UK due to Hypoderma were
estimated to be approximately E l 3 million (Tarry, 1986). The programme
was based on a combination of the voluntary use of pour-on organo-
phosphorus treatments, with compulsory treatment of cattle showing Hypo-
derma larvae in the spring, plus appropriate movement restrictions. The
programme was very successful and resulted in a decrease in the original
infestation levels from about 40% to less than 1% in four years. From 1982
it became compulsory to treat a whole herd in which Hypoderma was found
(Tarry,1986) and the programme was so effective that, recently, it has only
been possible to detect infestations by serological analysis of hosts
(Sinclair et al., 1990; Tarry et al., 1992) and even these revealed zero
positive in some 312 000 animals tested in 1992-1993 (Tarry, 1994).

However, as a result of relaxation of importation restrictions on cattle
from other EC countries, during May 1993 it was noticed that warbles
were present on some imported cattle: 42% of cattle groups entering
Britain tested positive for active infections (Tarry,1994); all of a sub-
sample were identified as H . bovis (D.W. Tarry,personal communication).
If any of the imported infestations matured before treatment and produced
adults that mated and laid eggs, then this will represent a severe set-back to
the eradication programme just at the time it was reaching success.
The dangers of a rapid reinfestation by Hypoderma were emphasized
by Scholl et al. (1986) in a report on progress in the insecticide-based
. Joint United States-Canada Cattle Grub Project. In a three-year period,
1983-1985, this project reduced populations of grubs by over 90%. Cattle
grubs are estimated to cause annual losses (excluding control) of over
US$600 million in the United States (Drummond et al., 1981). The effects
are mainly due to a loss in hide value and meat trim at slaughter; the direct
effects of infestation on weight gain and feed utilization of beef cattle are
negligible (Scholl et al., 1988).
Details of the infestation levels of cattle with Hypoderma in Europe,
which can be found in Gasca et al. (1992) and Losson et al. (1993), indicate
the importance of the parasite, for example, with 91% of farms ( n = 78)
surveyed in Belgium testing positive for infestations (J.F. Lonneux,
B. Losson, and L. Pouplard in Gasca et al., 1992).
The status of bot and warble flies in Sweden has been reviewed by
Andersson (1988): Hypoderma tarandi and Cephenemyia trompe are still
common on reindeer. In Canada, these two species have a significant effect
on caribou activity budgets as they increase the time spent standing and
moving and decrease the time spent feeding and resting (Downes et al.,
1986). In the Magadan region of the far east of the former USSR, over half
a million reindeer were treated with insecticide three times a year to protect
against these two species: 23% of deer hides were downgraded due to
warble fly damage (Shumilov and Nepoklonov, 1983).
. Hypoderma tarandi is considered one of the most economically signifi-
cant of reindeer parasites in Alaska, with hides of adult reindeer having up
to 2000 warble scars (Washburn et al., 1980). The effects of H . tarandi on
reindeer are summarized by Karter et al. (1992). They showed that, under
controlled conditions in the laboratory, H . tarandi oviposit close to the base
of newly grown hairs and that newly hatched larvae show a positive
thermotaxis. Both these behaviours would, in the wild, promote near
maximum hatchability, a short incubation period and high transmission
efficiency. In northern Norway, Folstad er al. (1991) recorded 99.9%
prevalence of infection of reindeer with H . tarandi, the larval burden
ranging up to 432 larvae per animal. The mean number of parasites per
MYIASIS OF HUMANS AND DOMESTIC ANIMALS 277
herd member was negatively correlated with the distance between calving
and summer grazing grounds. The calving grounds are where the greatest
larval shedding occurs. Folstad et al. (1991) hypothesize that the annual
migration between these two grounds is a behaviour that confers a reduc-
tion in Hypoderma levels, and that, in addition to predator avoidance and
enhanced access to good food, such parasite avoidance behaviour may have
played an important role in forming migration patterns among herbivore
populations.
Hypodermosis in man most frequently features skin allergies accompa-
nied by blood eosinophil differential counts varying from subnormal to
6 0 8 above the normal (Boulard and Petithory, 1977). The severity of
infection varies with the site of the larvae, from a “creeping myiasis”
caused by subdermal migrations (Uttamchandani et al., 1989), to ophthal-
momyiasis interna resulting in visual loss (Edwards et al., 1984), to rare
intracerebral myiasis (Kalelioglu et al., 1989). Since 1982 (Syrdalen et al.,
1982), H . tarandi has been increasingly reported as a cause of more or less
severe ophthalmomyiasis in humans, infestations even leading to blindness
(Kearney et al., 1991). Surgical techniques for removal of larvae in cases
of human ophthalmomyiasis are discussed by Syrdalen et al. (1982) and
Rapoza et al. (1986).
Larvae of Przhevalskiana burrow into the skin of goats in the same
manner as the larvae of Hypoderma in cattle (Puccini et al., 1987).
Infested animals lose condition and the perforation of the hide causes
considerable losses to its value. A good correlation has been found
between the loss of weight of young goats and the numbers of larvae of
P. silenus harboured by them (Liakos, 1986). The most recent studies of P.
silenus have been made in southern Italy, by Tassi et al. (1989) who also
review the literature on this species. They recorded infection rates ranging
from 30 to 81% of goats in a herd, each with up to a mean of 5.3 warbles:
there was a greater prevalence of infection among younger animals which
showed a higher mean intensity of infection.
4.1.4. Cuterebrinae
Dermatobia hominis occurs from Mexico (24-26”N) south throughout all
countries of central and south America, except Chile (Roncalli and Usher,
1988; Uribe et al., 1989), to northern Argentina (30-32”s). The annual
costs of D. hominis infestation (meat, milk and hide production losses)
were estimated to be some US$200 million annually in 1976 (Steelman,
1976): more recent estimations are needed. In Costa Rica up to 42% of
cattle are infested by D . hominis and up to 62% of water buffaloes (Sancho
et al., 1989). Thomas (1988) recorded a mean monthly infestation rate of
23% in cattle in the southern coastal Yucatan Peninsula of Mexico, with

71% of cattle being infested at least once during the wet season.
An indication of the potential effect of D . hominis on weight gain in
cattle is given by the study of McMullin et al. (1989): cattle treated with an
ivermectin bolus gained significantly more weight than controls which
received a placebo bolus (mean of 81 kg versus 47 kg). The increased
weight gain of the treated group may have been partly due to control of
other parasites as well, but for much of the trial the treated group had zero
live larvae whereas the untreated group had about 40-60 larvae. Female
larvae are probably a greater parasitic burden to hosts than males because
they take a longer period to reach maturity than male larvae and reach a
greater mature weight (Ribeiro and Oliveira, 1983; Lello er al., 1985).
The biology and economic importance of D . hominis have recently been
reviewed by Sancho (1988). A considerable amount of attention has been
paid to the location on the host where the nodules of Dermatobia are most
frequently located. In a study of Canchim breed of cattle (5/8 Charolais +
3/8 Zebu) in Brazil, Oliveira (1991a) found that 57% of Dermatobia
nodules were located on the left side of the body. He suggested that the
hosts’ preference for resting on their right side might be the reason for this
asymmetric distribution of nodules: 55% rested on their right side so
exposing their left side to carriers of Dermatobia eggs. The importance
of the protective value of the tail in influencing the location of nodules was
also emphasized. Hence, tail swishes can reach some 41% of the body
surface and only 16% of Dermatobia nodules were found in that, mainly
posterior, region (Oliveira, 1991~).Thomas (1988) also considered that the
activity of the tail may cause the lower number of nodules observed on the
hindquarters.
The ecology of free-living stages of D. hominis, third instar larvae, pupae
and adults, has been most recently studied in detail by Oliveira (1991b, d)
in Brazil. He observed that the duration of pupal stages was temperature
dependent, ranging from 62-70 days in the coldest months (mean tempera-
ture 17-19°C) to 34-37 days in the warmest (and wettest) months (mean
temperature 22-24°C). The emergence rate was greater in the hotter, humid
months than in the colder, drier months (range 15-52%). Longevity of
adults was also linked to temperature, being longest in the coldest months
(up to 19 days), but humidity was also important, thus in the hottest months
longevity was longer when precipitation was greater. These biological
parameters combine to give greatest frequencies of cattle infestation with
Dermatobia in warm and humid months and least in cold and dry months
(Oliveira, 1991d). Similar results were found by Ribeiro et al. (1989).
Recent research has also provided considerable information on the egg
stage of Dermatobia. The eggs are attached firmly to the body of the carrier
by an adhesive that does not appear to lose its integrity in museum speci-
mens for over 50 years (Cogley and Cogley, 1989); the eggs are also
strongly adhered to each other. The adhesive(s) can be broken down by
enzymatic activity (papain and bromelain). The combination of egg-to-egg
and egg-to-carrier attachment increases the likelihood of the eggs remaining
attached and increases the overall rigidity of the egg mass. Cogley and
Cogley (1989) concluded that a number of structural and location features
enhance the ability of larvae to infest a host, e.g. the ventral placement of
eggs such that the hatching end is nearest the host’s skin, the offsetting of
egg tiers which enables larvae to escape without obstruction from overlying
eggs, and egg curvature that helps to give all eggs a nearly equal contact
with the host.
In addition to its major importance as a veterinary pest, D . hominis is a
common pest of humans. Rarely it can cause fatalities, as when larvae
penetrate the fibrous portion of the bregmatic fontanel of infants (Rossi and
Zucoloro, 1972). As air travel increases, cases of human infestation with
D. hominis are recorded more frequently outside its natural range. Recent
reports in the literature have included cases not only in the New World (e.g.
USA: McIntyre, 1989; Lowry, 1992) but also in the Old World, in Belgium
(Deroo et al., 1990), England (Hay, 1990), France (Nderagakura et al.,
1989), Italy (Polidori et al., 1992), Poland (Wegner et al., 1986, 1992),
Japan (Maeda et al., 1990) and Saudi Arabia (Qadri and Al-Ahdal, 1988). In
addition to passive migration in humans, larvae of both D. hominis
(Bourdeau et al., 1988; Roosje et al., 1992) and Cordylobia anthropophaga
(Fox et af., 1992) have also been imported to Europe with dogs.
In contrast to Dermatobia, Cuterebra species have a virtually negligible
economic impact as they naturally parasitize rodents and lagomorphs
(Catts, 1982). However, they can parasitize humans in North America, as
reported in detail by Baird et al. (1989). Alouattamyia baeri is a cuterebrid
parasite of howler monkeys (Catts, 1982), but can cause a pulmonary
myiasis of man (Fraiha er al., 1984).
4.2. Calliphoridae
4.2.1. Cochliomyia
The distribution of New World screwworm fly, Cochliomyia hominivorax,
extends from the southern states of the USA through Central America and
the Caribbean Islands to northern Chile, Argentina and Uruguay. In North
America the fly used to spread north and west each summer into more
temperate zones from its overwintering areas near the USAhlexican
border. The fly was of greatest significance as a pest of livestock, neces-
sitating the continued costs of vigilance, treatment and control. In the
epidemic year of 1935 in Texas there were approximately 230 000 cases in
livestock and 55 in humans (Dove, 1935). Up to 1958, the annual cost of C.
hominivorax control in the United States was estimated to be US$140
million. Large-scale screwworm fly control, by sterile insect technique
(SIT), was initiated in the south eastern states of the United States of
America in 1957-59. Subsequent control operations spread the area of
sterile male release and in 1966 effective control of C. hominivorax in
the US was declared. Several outbreaks since then, most notably in 1968
and 1972, occurred but control was quickly reimposed and no cases of
infestation have been recorded since 1982. The eradication programme has
subsequently been directed against the fly in Mexico, Puerto Rico, Vieques
* and the Virgin Islands (Graham, 1985; Krafsur et al., 1987). SIT has been
advocated for use on other islands in the Caribbean region where it can be a
serious pest of livestock (Rawlins and Mansingh, 1987) and humans
(Rawlins, 1988).
In 1988, C. hominivorax were discovered in an area 10 km south of
Tripoli in Libya (Gabaj et al., 1989). This was the first known established
population of this species outside the Americas. The fly quickly spread to
infest about 25 000 km2. In 1989 there were about 150 cases of myiasis by
C. hominivorax, but in 1990 a total of 12 068 confirmed cases of screw-
worm fly myiasis were recorded and, at its peak, almost 3000 cases were
seen in the single month of September 1990, mainly in sheep. Humans
were also affected (El-Azazy, 1990; Reichard et al., 1992). It was esti-
mated that if unchecked the infestation could cost the Libyan livestock
industry about US$30 million per year and the North African region
approximately US$280 million per year (Lindquist et al., 1992). This led
to the implementation of a major international control programme to
eradicate the fly from this area (see section 6.4.2).
. Cochliomyia macellaria is a ubiquitous carrion breeder in the Americas,
but can act as a secondary invader of strikes, and is known as the
secondary screwworm fly. It can cause myiasis in humans, usually in
immobile or debilitated persons (Smith and Clevenger, 1986). A rare case
of Cochliomyia minima myiasis has been reported in a dog in Puerto Rico
(Lebn and Fox, 1980).
4.2.2. Chrysomya
The Old World screwworm fly Chrysomya bezziana screwworm is an
obligate parasite which occurs throughout much of Africa, India, the
Arabian peninsula, southeast Asia and the Indonesian and Philippine
islands to New Guinea (Norris and Murray, 1964; Spradbery and Kirk,
1992). The precise status of C. bezziana as a clinical and economic pest is
uncertain, particularly in sub-Saharan Africa, and few studies have been
able to obtain quantitative estimates of myiasis incidence, its clinical or

economic importance. The absence of livestock throughout much of its
range in sub-Saharan Africa, due to the presence of trypanosomiasis and
its vector the tsetse fly (Glossina spp.) may have substantially limited its
economic impact (Hall, 1991). However, C. bezziana has been inadvertently
introduced into several countries in the Middle East and such an introduc-
tion is believed to pose a major economic threat to the pastoral industry of
Australia (Rajapaksa and Spradbery, 1989; Sutherst et al., 1989) where it
has been estimated that its cost to the livestock industry would be up to
A$430 million at 1990 values (Spradbery, 1994).
Chrysomya megacephala is a native of Australasian and Oriental
regions, known to act as an agent of myiasis in domestic animals and
humans (Baumgartner and Greenberg, 1984). It is anautogenous and is
commonly known as the ‘Oriental latrine fly’ because it can breed in
faeces as’ well as on carrion. It has been introduced inadvertently into
Africa and also into the New World (for reviews see Olsen et al., 1993):
it entered Brazil around 1975 along with Chrysomya putoria and C .
albiceps (Guimarges et al., 1978). These species have dispersed rapidly
to reach Central and North America (Wells, 1991).
The hairy maggot blowfly, Chrysomya rufifacies, is a tropical Australa-
sian and Oriental species. It is morphologically similar and closely related
to the African species Chrysomya albiceps. Erzinqlioglu (1987) proposed a
character on dorsal process 1 for separating the larvae of these two species.
However, Tantawi and Greenberg (1993) proposed instead other characters
on the processes and urged caution in using previously accepted characters,
for larvae and adults, especially when dealing with small numbers of
specimens. Both C. rufifacies and C . albiceps are saprophagous, normally
laying batches of 200 eggs on carcasses, but they may also act as faculta-
tive ectoparasites. In Australia and New Zealand C. rufifacies is predomi-
nantly a summer species and in parts of its range it may be the dominant
species in carcasses (Anderson et al., 1988). Chrysomya rufifacies may be
capable of initiating sheep myiasis, but more commonly it will act as a
secondary invader of myiasis in Australia (Anderson et al., 1988) and New
Zealand (Tenquist, 1977; Heath and Bishop, 1986). It has been reported as
a serious pest of newborn calves in Hawaii, with up to 10% mortality,
mainly of calves contaminated with placental and foetal tissues after birth
(Shishido and Hardy, 1969). First instars are entirely necrophagous but
second and third instars may be facultatively predaceous on other dipteran
larvae. This may give it considerable competitive advantage in crowded
carcasses and in sheep myiasis, where C. rufifacies may also act as a
biological control agent, repelling and killing larvae of L. cuprina. In
1978, C . rufifacies was introduced into Central America from where it
has dispersed into the southern states of the USA (Baumgartner and
Greenberg, 1984). Because of its predaceous behaviour, it may reduce

natural populations of Cochliomyia macellaria (Wells and Greenberg,
1992). The biology of C . rujfacies has recently been comprehensively
reviewed (Baumgartner, 1993).
4.2.3. Lucilia
Twenty seven species have been described within the genus Lucilia, found
worldwide, although they are originally and predominantly Palaearctic and
Ethiopian in distribution (Aubertin, 1933). However, only two, Lucilia
sericata and Lucilia cuprina, are of major clinical and economic signifi-
cance as primary agents of cutaneous myiasis, particularly affecting sheep,
although they may also strike a range of other wild and domestic animals
and humans.
Cases of blowfly strike of sheep have been recorded from many parts of
nothern Europe where the primary species responsible is L. sericata. Sheep
strike by L. sericata has been recorded in the Netherlands (Baudet and
Nieschultz, 1938) and in Scandinavia (Rigndahl, 1942). In the summer of
1981 in north and west Germany, myiasis by L. sericata resulted in sheep
mortality rates of up to 10% (Liebisch et al., 1983). Lucilia sericata was
shown to be the most important primary agent of sheep myiasis in Scotland
(Ratcliffe, 1935; Haddow and Muirhead Thompson, 1937), Wales (Davies,
1934) and the Ukraine (Mashkei, 1990).
In a comprehensive national survey of sheep blowfly strike in Britain,
MacLeod (1943) found that L. sericata was by far the most important
primary myiasis fly; only in the western highlands of Scotland was it
found in less than 90% of all strikes. However, the Lucilia caesar group
(L. caesar and L. illustris), Protophormia terraenovae, and the bluebottles
Calliphora vicina and C . vomitoria were also found in small numbers,
though rarely in pure cultures, suggesting that they generally act only as
secondary agents of myiasis. In a more recent, though considerably less
detailed, study than that of MacLeod, samples of larvae were collected
'from strikes at spring shearing in England and Wales. The same general
pattern of species in myiases was confirmed, with 81% of strikes composed
of L. sericata alone, 13% of mixed cultures of L. sericata and L. caesar and
6% of L. caesar alone. No other species were found (Wall et al., 1992a).
Larvae of L. sericata in pure cultures have been collected from strikes on
domestic rabbits, dogs, cats and wild hedgehogs and birds which had been
brought into veterinary surgeries in the UK (Wall, unpublished data).
However, the extent to which existing injury predisposed these animals
was unknown. A pure culture of L. illustris has recently been recovered
from myiasis of a fox cub (Vulpes vulpes) in south west England (Wall,
unpublished data).
A pronounced geographic trend was evident in the data collected by

MacLeod (1943): P. terraenovae was restricted to strikes in the north and
west of Britain and a similar, though less marked, trend could be seen with
L. caesar and the Calliphora species which were relatively rare in strikes in
southern Britain or Ireland (Table 6). The occurrence of L. caesar in strikes
in the more northerly and westerly areas of Britain is supported by a report
that, in Norway, in 27 cases of sheep myiasis the primary species was
found to be L. caesar, occurring alone or in combination with L. illustris
(Brinkmann, 1976).
Myiasis caused by L. sericata and L. cuprina was reported to have
affected 0 . 2 6 4 6 5 % of sheep in the wooded zone of the Ukraine and
0.14.4% in the steppe zone, with 27-32% mortality among affected sheep
(Mashkei, 1990). In England and Wales, a questionnaire survey of sheep
farmers showed that in 1989-90 blowfly strike affected over 80% of farms,
where an.average of 1.5% of sheep were struck each year (French et al.,
1992). This prevalence equates to about half a million sheep struck
annually, from a national flock of about 30 million. Nevertheless, examina-
tion of the literature indicates that 50 years ago strike prevalence in Britain
was considerably higher, with between 10 and 20% of sheep struck. The
primary change in the intervening 50 years has been the development and
use of powerful organochloride and organophosphorus insecticides. In the
absence of such control, strike rates in northern Europe might be expected
to return to the levels seen in the first half of this century.
The status of the blowfly Lucilia cuprina (Wiedemann) as an agent of
primary myiasis in Australia was first correctly established in about 1930
(Norris, 1990) and subsequent surveys have confirmed L. cuprina as the
dominant sheep myiasis species for mainland Australia (Mackerras and
Fuller, 1937; Watts et al., 1976) and Tasmania (Ryan, 1954), being present
in 90-99% of flystrike cases (Dalwitz et al., 1984). Lucilia sericata is
present in Australia but is generally confined to more urban habitats. It can
and does act as a primary myiasis fly however. In many parts of Australia
L. cuprina is able to breed throughout the year but, in the main sheep areas
of S.E. Australia there is usually a long winter period of low fly abundance.
Today, L. cuprina remains a major pest in Australia and up to 3 million
sheep may be killed each year. Its annual cost to the Australian sheep
industry through control and production losses are estimated at around
A$150 million (Beck et al., 1985).
Lucilia sericata arrived in New Zealand over 100 years ago (Miller,
1939) and for many years was the primary myiasis fly in this country,
occurring in 75% of all cases of sheep strike (Heath and Bishop, 1986).
Strike by L. sericata is most prevalent between March and April. Other
species, including Calliphora stygia and Chrysomya rujifacies are also
important in New Zealand (Heath and Bishop, 1986; Dymock et al.,
Table 6: Agents of sheep myiasis in Britain. The prevalence of Lucilia sericata, Lucilia caesar group (L. caesar or L. illustris),
Protophormia terraenovae and Calliphora spp. in pure and mixed cases of sheep myiasis in Britain (Pure = percentage of the total
number of samples identified (n) that contained pure cultures of the fly species; Total = percentage of total number of samples in
which the fly species was found, in mixed and pure cultures). Data from MacLeod (1943).
Protophormia
Lucilia sericata Lucilia caesar terraenovae Calliphora SQQ.
n Pure Total Pure Total Pure Total Pure Total
N.W. Scotland 19 79 100 5 16
Outer Hebrides 11 100 100
W. Highlands 107 44 73 16 38 7 18 1 7
Central Highlands 33 64 91 3 15 18 6 6
N.E. lowlands 17 100 100
Central lowlands 133 77 94 1 14 4 7
Tweed Basin 81 96 100 2 1
Solway and Lake District 117 78 98 15 6 1 4
Pennines and Yorkshire 26 81 96 11 4 4 4
North Wales 145 85 94 4 13 1 1
South Wales 59 98 98 2 2
Central England 49 100 100
Eastern England 33 97 100 3
Southern England 39 92 100 8
S.W. Peninsula 65 98 100 1
Ireland 97 98 100 2
1991). In 1976, a questionnaire survey in New Zealand showed that the

annual prevalence of flystrike in sheep was 1.7% in the North Island and
0.7% in the South Island and the annual cost to farmers was at least $1.7
million (Tenquist and Wright, 1976). However, in the early 1980s L.
cuprina was first discovered in New Zealand and now, despite its low
abundance (Dymock and Forgie, 1993), it appears to be displacing L.
sericatp, to become the most important primary cause of flystrike in sheep.
In southern Africa the primary myiasis fly of sheep is L. cuprina
(Waterhouse and Paramonov, 1950). Although this species had been
known in South Africa since 1830, little sheep strike was recorded until
the early decades of the 20th century, possibly as a result of the introduc-
tion of more susceptible Merino breeds, changes in husbandry practices
and/or the development of more aggressive fly strains (Norris, 1990).
Lucilia sericata and L. cuprina (= L. pallescens of some authors for North
America) are present in areas of North America (Hall and Townsend, 1977)
but little appears to be known about their clinical or economic importance
as agents of myiasis there. In 1991, L. cuprina was collected for the first
time in Europe, among a sample of calliphorids from Zaragoza, Spain
(Rognes, 1994). How long the species has been present in Spain and its
veterinary importance there are presently unknown.
Lucilia species can cause a facultative myiasis of humans, particularly
the elderly or debilitated, with cases reported for L. sericata (UK, Roche et
al., 1990; USA, Greenberg, 1984) and L. cuprina (Australia, Lukin, 1989).
4.2.4. Phormia, Protophormia

The most im.portant species are Phormia regina and Protophormia
terraenovae. Both are similar in habits, usually breeding in carrion, but
both can cause myiasis, particularly for P . regina in North America (Hall,
1948; Hall et al., 1986). Protophormia terraenovae is a Holarctic, cold-
loving species. It is abundant in early spring in Finland (Nuorteva, 1987)
and is the dominant blowfly in the Arctic and subarctic (Vinogradova,
1986). As shown in MacLeod’s (1943) study, P . terraenovae is more
important in sheep strike in Britain in northern Scotland (Table 6).
4.2.5. Calliphora
There are a great many species in this widely distributed genus, particularly
in the Holarctic and Australasian regions. Species of Calliphora are
primarily carrion feeders (e.g. Davies, 1990) but a number can act as
secondary or tertiary agents of myiasis (Zumpt, 1965). The two most
important species in the northern hemisphere are Calliphora vicina and
Calliphora vomitoria. Attempts to induce sheep strike by C . vicina proved
unsuccessful (MacLeod, 1937) and led MacLeod to conclude that this

species was physiologically unable to strike sheep. He suggested that
this was either because the sheep body temperature was fatally high or
because larvae were unable to feed on the animal tissues without the prior
activity of Lucilia larvae. Nevertheless, C . vicina has been recorded as
laying eggs on living small mammals (ErzinClioglu and Davies, 1984).
The Western Australian brown blowfly, Calliphora albifrontalis and the
lesser brown blowfly, Calliphora nociva, are also important native species
found in sheep strike in the Australasian region (Anderson et al., 1984). In
Western Australia C . albifrontalis may be responsible for up to 10% of
single-species strikes. In New Zealand Calliphora stygia may be a common
secondary invader of ovine myiasis (Heath and Bishop, 1986) being present
'in strikes from October to May.
4.2.6. Cordylobia
As with Dermatobia, human movements carry infestations of Cordylobia
anthropophaga outside the endemic areas with increasing frequency, for
example, in Belgium (Hausdorfer-Scheiff et al., 1993), France (Gall et al.,
1987), Italy (Pampiglione et al., 1993), the UK (Chopra et al., 1985), Japan
(Kagei et al., 1989) and the USA (Ockenhouse et al., 1990). Of interest in
this regard is the acquisition of Cordylobia myiasis in northern Europe by
two brothers who had never been to Africa (Baily and Moody, 1985): the
explanation for this infestation was that their father, who had made several
recent trips to Africa, might have brought back eggs of the tumbu fly with
his baggage. More difficult to explain is the case of a British woman who
had also never been to Africa, but had acquired an infection with tumbu fly
in Spain (Laurence and Herman, 1973). It appears that C . anthropophaga
may not be restricted to sub-Saharan Africa as is usually stated (Zumpt,
1965): seven cases were recently reported originating in the Asir region
of southwestern Saudi Arabia (Omar and Abdalla, 1992).
A rare case of human myiasis due to C. rodhaini was reported from Italy,
in a patient returned from Ethiopia. The case was particularly severe
involving some 150 larvae (Pampiglione e f al., 1991).
4.3. Sarcophagidae
The most important genus acting as agents of myiasis in this family are
Wohlfahrtia, the key species being W. magnijica. It is considered to be the
most important myiasis-causing species of camels (Higgins, 1985), but is
even more important as a pest of sheep. Levels of infestation appear to be
higher in sheep in Eastern European countries (e.g. Bulgaria, 23-41%,
Nedelchev, 1988; Stavropol area of the former USSR, 30-50%, Pokidov

and Goncharov, 1971), than those observed in the Mediterranean basin (e.g.
Spain, 0.7-8.5%, Ruiz-Martinez et al., 1987, 1991; Israel, ca 1.5-12.5%,
Hadani et al., 1971). Exceptionally, a report from Romania states that
8&95% of sheep were infested, with 20% fatalities of newborn lambs
(Lehrer et al., 1988). This high incidence is thought to be a consequence of
the introduction and massive increase in numbers of sheep originating from
Australia and New Zealand (Lehrer and Verstraeten, 1991).
Portchinsky (1916) recorded many cases of human infestation with W.
mugn$ca from Russia, with approximately 80% in children of 10 years or
less and approximately 70% affecting the ears. Most other cases affected the
head area, either eyes, mouth or nose. Human infestations due to W.
magn$ca are still recorded, from the ear (El-Kadery and El-Begenny,
1989), eye (Baruch et al., 1982), nose or mouth (Zeltser and Lustmann,
1988). .
5. PHYSIOLOGY OF MYIASIS
5.1. Predisposing Conditions for Myiasis
5.1.1. Oestridae
Maia and Guimaries (1985) reported an association between abscesses on
cattle and infestation by Dermatobia hominis which can be explained by
the observations of Koone and Banegas (1959) that such abscesses
(including ongoing Dermatobia infestations) are attractive to important
egg vectors such as Sarcopromusca arcuata. Lesions of D . hominis on
cattle are not attractive to adult Cochliomyia hominivorax and therefore do
not usually become secondarily infested by screwworms (Thomas, 1987).
However, myiasis due to C. hominivorax was observed in some sheep after
removal of Dermatobia larvae (Amarante et al., 1992). In Sao Paulo State,
Brazil, sheep were affected by D. hominis after shearing, particularly in the
scrotum of rams (Amarante et al., 1992).
Within a single breed of cattle there may be considerable differences in
the rate of infestation of individuals by Dermatobia hominis (e.g. Maia
and Guimaries, 1985) and there is evidence for differences in breed
susceptibility. Steers of Bos indicus had significantly fewer naturally
acquired nodules of D. hominis than did steers of Bos taurus (14.6 versus
21.5 nodules, respectively; Moraes et al., 1986). Similarly, Oliveira and
Alencar (1992) showed that crosses of Holstein-Fresian and Guzera breeds
were increasingly susceptible to Dermatobia infestation as the proportion
of Holstein-Fresian increased. However, Thomas (1988) showed no dif-

ference in susceptibility of Zebu and Brown-Swiss cattle in Mexico. The
question of differential host susceptibility and its basis needs much further
study in controlled conditions.
5.1.2. Calliphoridae and Sarcophagidae

For the primary screwworm flies, Cochliomyia hominivorax and
Chrysomya bezziana, eggs are laid in pre-existing wounds or in body
orifices, such as eyes, ears or nostrils. Even wounds the size of a tick bite
are reported to be sufficient to attract oviposition. The precise semiochemi-
cal and chemotactile cues causing attraction and leading to oviposition are
unknown, but wound fluids and blood are known to be attractive (Hammack
and Holt, 1983). Bacterial metabolites also may increase the attractiveness
of screwworm infested wounds as oviposition sites (Humphrey et al., 1980;
Hammack et al., 1987).
The key factors determining the incidence of flystrike in sheep by Lucilia
are sheep susceptibility and fly abundance, both of which are affected by a
range of environmental and management variables (Wardhaugh et al.,
1989; Wardhaugh and Morton, 1990; French et al., 1994a). Blowfly strike
by L. sericata occurs most commonly in the breech region and is strongly
associated with faecal soiling (Leiper, 1951). Faecal soiling is also an
important predisposing factor for strike by L. cuprina (Watts et al.,
1978), as it is for breech myiasis in sheep caused by W. magnifica
(Nedelchev, 1988). The role of odours in the attraction to and subsequent
oviposition of L. sericata and L. cuprina on sheep has been reviewed
recently (Ashworth and Wall, 1994).
Body strike, strike to the back, flanks and withers, is frequently the main
form of myiasis caused by L. cuprina in Australia (Watts et al., 1979).
Body strike occurs most commonly around the shoulders and back region
and is frequently associated with the incidence of bacterial dermatophilosis
(Wardhaugh and Dallwitz, 1984). Dermatophilosis is a chronic bacterial
infection, caused by the bacterium Dermatophilus congolensis which
invades the epidermis (Roberts, 1967). Body strike in Australia is more
often associated with bacterial fleece rot, a superficial dermatitis induced
by moisture and bacterial proliferation on the skin, resulting in a matted
band of discoloured fleece. It is caused predominantly by Pseudomonas
aeruginosa (Watts er al., 1979). It is possible that dermatophilosis and
fleece rot act synergistically in attracting blowflies and their subsequent
oviposition (Gherardi et al., 1983). There is little recorded involvement of
either form of dermatitis in predisposing sheep to strike in northern Europe.
Genetic variation in fleece rot and body strike susceptibility have been
identified between Merino sheep strains, bloodlines and between individual
sheep within flocks (Raadsma, 1987; Raadsma et al., 1989). There is,
therefore, the potential for selection for resistance to reduce the incidence
of this condition. In practice it is likely that indirect selection criteria, such
as for resistance to fleece rot, would be employed rather than direct
selection for resistance to body strike (Raadsma, 1991).
The risk of myiasis by L. sericata in England and Wales has been shown
to increase with increasing flock size and stocking density and to decrease
with increasing farm altitude and latitude (French et al., 1994a). In
Australia, flystrike was shown to be positively related to increases in the
density and activity of gravid L. cuprina, rainfall, cloud cover and the rate of
pasture growth (Wardhaugh and Morton, 1990). The analysis suggested that
rainfall determined overall levels of strike, whereas pasture conditions and
cloud cover determined the type of strike, with crutch strike replacing body
strike under dry conditions and when fly densities were low (Wardhaugh
and Morton, 1990).
Calving seemed to predispose camels to myiasis by Wohlfahrtia magni-
frca, gravid female flies being attracted to lochial fluids and damaged
tissues (Hadani et al., 1989).
5.2. Pathology and Immunology
5.2.1. Oestridae
The parasitic rhinitis caused by the larvae of Oestrus ovis is characterized
by a sticky and mucoid nasal discharge, at times haemorrhagic (Roncalli,
1984b). Histopathological changes in the nasal tissues of sheep and goats
due to infestation by 0. ovis were recorded by Jagannath et al. (1989a):
they were characterized by catarrh, infiltration of inflammatory cells and
squamous metaplasia, with conversion of secretory epithelium to stratified
squamous type. Yilma and Dorchies (1993) reported a significant reduction
in the population of 0. ovis artificially placed into sheep nostrils, which
they suggested was due to host immune reactions. Previously, Rogers and
Knapp (1973) had demonstrated levels of mortality of the immature stages
of 0. ovis ranging from 9699%. There is a danger in ovine oestrosis of
secondary infections leading to lung abscesses (Dorchies et al., 1993).
Harvey (1986) presents a comparative study of 30 cases of human
ophthalmomyiasis due to Oestrus ovis. The common symptom of all was
an acute conjunctivitis, 63% also showed lid oedema and 43% a superficial
punctate keratopathy,produced by movement of the larva across the cornea.
However, in rare cases the larva may invade the interior of the eye,
producing extensive retinal destruction and preretinal fibrosis (Rakusin,
1970).
Hussein et al. (1982) describe the pathology of infestations of camels

with Cephafopina titillator, which includes congestion of the nasal cavity
with mucus, severe inflammatory and degenerative changes, leading to
extensive damage of nasopharyngeal tissues, and the formation of
lymphoid nodules at the site of larval attachment in the pharynx.
The pathological effects of Hypoderma in cutaneous lesions are well
known (Scholl, 1993) but, until recently, the pathological changes asso-
ciated with migrating larvae were less well known. Panciera et al. (1993)
have now described effects of first instars of H . lineafum on the connective
tissues through which they migrate. These are characterized by yellow or
greenish gelatinous, oedematous areas with an overwhelming eosinophil
infiltration.
It is surprising that perhaps the most detailed account of tissue changes
accompanying infestation of mammals by larvae of Cuterebridae concerns
infestation of rodents, white-footed mouse (Peromyscus leucopus feuco-
pus) by Cuterebra angustifrons, rather than of livestock by Dermatobia
hominis (Payne and Cosgrove, 1966). The lesions made by this bot fly in
mice are similar to those of D . hominis in cattle, involving a cavity in the
loose subcutaneous tissue, not usually extending into the underlying
muscle. There is a thin layer of necrotic tissue next to the larvae and the
bot feeds on tissue debris and exudates. Following departure of the bot,
healing and closure of the lesion can be very rapid, within nine days for
C . angustifrons and five days for C . beameri in pack rats (Beamer et al.,
1943).
Although many authors have reported the clinical and histological
features of human infestations with Dermatobia hominis (e.g. Lane et al.,
1987), there are few accounts of immunological features. Grogan et al.
(1987) report that there is a complex host immunological response to the
larva, including lymphocytes, eosinophils, activated fibroblasts, mature
histiocytes, mast cells/basophils, plasma cells and Langerhans cells. A
similar set of cells is found in mice infested by Cuterebra angustifrons
(Payne and Cosgrove, 1966). Immunohistochemistry revealed that the
. dominant dermal cells responding to the larva were activated (Ia') T-helper
cells which, in turn, probably stimulate plasma cell production and B-cells
to produce antibody (Grogan et al., 1987). The activated fibroblasts pro-
duce collagen that may be important in containment of the larva within its
fibrous, subcutaneous nodule. Grogan et al. (1987) could not explain how
the larva escaped immunological destruction. However, they only examined
a punch biopsy of the skin lesion at about one month after infestation. In
future studies, examination of lesions produced by D . hominis over the
whole period of infestation would be of value. Progress towards solving
this problem comes from the work of Lello and Boulard (1990), who have
studied the immune responses of rabbits to experimentally produced
infestations with D . hominis. They showed that the larvae of this species
elicit a humoral response from the host. The first instar larvae produced the
greatest host reaction and allowed the earliest detection of infestation by
ELISA. However, the initial immune response against antigens produced
by second and third instar larvae was depressed during the course of an
infestation, but started to increase as soon as the larvae left their host. This
suggests that immunosuppression may be a phenomenon of Dermatobia
infestation. Weisbroth et al. (1973) demonstrated that the antigens
provoking the immune response in rabbits naturally infected with Cuterebra
buccata resided in the alimentary tract and haemolymph fractions of
dissected larvae and that sensitization of the host occurred as a consequence
of exogenous larval secretions injected during feeding.
Dermatobia infestations also produce a cell-mediated immune response
(Lello and Peraqoli, 1993). Since immunization of rabbits with a crude
antigen of D . hominis led to higher levels of cellular response and antigen-
specific antibody production, vaccination against first instar larvae may be
a future means of biological control of this myiasis.

During the 6-7 days of larval feeding, Chrysomya bezziana burrow deeply
into the host’s tissues so that only the posterior segment and spiracles of the
larvae are exposed. The same is true of infestations with Cochliomyia
hominivorax and Wohlfahrtia mugnijica. In cattle, infestation by C .
bezziana has been described as causing intermittent irritation and pyrexia,
followed by the production of a cavernous lesion. The tissue shows pro-
gressive liquefaction, necroses and haemorrhage, before the larvae leave
the wound. Histologically, two distinct phases are observed: the first being
intense neutrophil infiltration and haemomhage associated with the growth
of the larvae; the second being a fibroplastic healing phase in which mast
cells and eosinophils are prominent (Humphrey et al., 1980).
Infestation of the perineal and vulvar areas of camels with Wohlfahrtia
magniJica results in marked deformity and fibrosis, with possible
complications in future calvings (Hadani et al., 1989).
In contrast to the feeding behaviour of screwworm larvae, the larvae of
Lucilia spp. usually feed superficially on the epidermis and lymphatic
exudate or on necrotic tissue. Only when crowded will they begin to
feed on healthy tissue. Digestion occurs extra-orally by means of amylase
in the saliva and proteolytic enzymes in the larval excreta. Feeding begins
with the head elevated and the mouth hooks in the abducted position. The
head is lowered until the grooved stoma1 disc contacts the feeding sub-
strate. The mouth hooks are then adducted and driven into the crevices of
the feeding substrate surface. Maceration of the substrate occurs as the
retractor and protractor muscles of the head contract and relax alternately,
while the mouth hooks are held in an adducted position. The semiliquid
material is then sucked through the oral aperture into the atrium (Barnard,
1977).
Sheep struck by L. cuprina display a rapid increase in temperature and
respiratory rate accompanied by loss of weight and appetite (Broadmeadow
et al., 1984). The animals become anaemic and suffer severe toxaemia,
with both kidney and heart tissues affected. A massive cellular infiltration
occurs in the skin of sheep within 48 h of primary or secondary infections
with L. cuprina, with a complex of cellular immune responses (Bowles et
al., 1992). The feeding activity of the larvae may cause extensive tissue
damage, which, in combination with the larval proteases produced (Bowles
‘et al., 1988), results in the development of inflamed, abraded or under-
mined areas of skin. This may result in considerable distress to the struck
animal, a loss of fertility (Heath et al., 1987) and, if untreated, rapidly leads
to death from chronic ammonia toxicity (Broadmeadow et al., 1984;
Guerrini, 1988).
Myiasis from a range of species has been shown to produce an immuno-
logical response in the host. Sheep struck by L . cuprina produce specific
antibodies in the serous exudate produced at the skin in response to the
feeding activity of larvae (O’Meara et al., 1991): sheep bred for resistance
to blowfly strike produce greater exudate protein release during infection
(O’Meara et al., 1992). Repeated exposure to four or five infestations of
these larvae at two-week intervals produces at least partial resistance to
reinfection (O’Donnell et al., 1981), but it is short-lived and requires
frequent larval exposure (Sandemann et al., 1992). Antibodies to whole
third instar larvae have been shown to be present in previously struck sheep
and significant mortality of larvae is observed in a challenged sheep, while
growth retardation is seen when larvae are cultured in vitro in the presence
of serum from previously infested sheep (Eisemann et al., 1990).
Larvae of Cordylobia anthropophaga rarely cause the severe pathology
in humans seen in infestations with Dermatobia hominis. They remain in
the skin or subcutaneous tissues and do not migrate into deeper tissues. The
skin surrounding the furuncles can become erythematous, oedematous and
tender to touch. An inflammatory infiltrate, consisting of lymphocytes,
histiocytes, neutrophils and eosinophils, extends throughout the tissues
around and below the larva (Ockenhouse er al., 1990). The larvae secrete
bacteriostatic fluid which can prevent secondary infections (Hausdorfer-
Scheiff et al., 1993). Blacklock and Thompson (1923) reported an acquired
immunity of dogs, guinea pigs, monkeys and humans to infestations by C.
anthropophaga and postulated at that early time that it might be possible to
immunize cattle against myiasis species, in particular Dermatobia and
Hypoderma.
6. NEW AND IMPROVED CONTROL TECHNIQUES
There are basically three levels at which control of myiasis species can be
considered:
1. Suppression or eradication of fly population (e.g. eradication of
Cochliomyia hominivorax and Hypoderma species);
2. Avoidance of infestation where adult control is not possible (e.g. by fly
screening, dressing of wounds, prophylactic treatments);
3. Treatment because of failure of both above levels (removal of larvae
. manually or by insecticides, application of antibiotics).
The location of the parasites for much of their developmental stages on
the host means that control techniques can be very precisely targeted
against at least the larval stage. The precision of targeting and the propor-
tion of the population that can be reached depends on the host specificity of
the parasite (i.e. whether there are wild animal, reservoir hosts which
cannot be easily treated), and its degree of dependence on the host (i.e.
whether or not it is a facultative species that can also develop on carrion).
6.1. Insecticides
6.1.1. Oestridae
An aerosol technique using trichlorphon has been developed against
Oestrus ovis (Ilchmann and Splistester, 1982) and has been used for
large-scale treatment of sheep in Mongolia (Ilchmann et al., 1986). How-
ever, most treatments for 0. ovis are based on systemics, applied by
injection or oral dosing. Thus, Rafoxanide administered orally at a dose
rate of 7.5 and 10 mg kg-' gave 94100% and 100% effective cure,
respectively, of 0. ovis in sheep (Horak et al., 1971; Roncalli et al.,
1971). Injected at a rate of 3 mg kg-', it gave 94100% reduction in
larvae (Arm er al., 1982; Schindler et al., 1986). Following a preliminary
epidemiological survey that demonstrated three generations of 0. ovis per
year in south-west France, a control regimen of two treatments of Closantel
(10 mg kg-') at eight-week intervals was tested by Dorchies et al. (1992)
and found to be very effective in improving sheep condition and protecting
against late infections. Previously, Closantel had been shown to be effec-
tive in both cure and prevention of 0. ovis infection: 98% of larvae were
eliminated in treated animals and there was a 75% reduction in incidence in
treated animals compared to controls after eight weeks (Dorchies et al.,
1989).
Ivermectin applied at a dose of 200 pg kg-' killed 99% (injection:

Puccini et al., 1983; Schindler et al., 1986) and 100% (orally: Roncalli,
1984b) of Oestrus ovis in sheep. When given as a subcutaneous injection at
the same dose, ivermectin also proved an effective treatment for the
condition associated with nasal infestations of camels by Cephalopina
tirillator (Sharma, 1992).
Both trichlorphon and ivermectin pastes administered to horses in
Hungary resulted in eviction of Gasrerophilus larvae: no larvae were
expelled by untreated controls (Egri, 1989). The major excretion of larvae
occurred 28-52 and 50-64 hours after treatment with trichlorphon and
ivermectin, respectively. Rastegayev (1988) used ivermectin against both
Rhinoestrus and Gasterophilus in horses, either as a 1.87% oral paste or a
1% injectable solution. Seven days after treatment, neither parasite could
be detected in injected horses. In orally treated horses, Rhinoestrus was
detected in only 6% and Gasterophilus in only 2%, whereas all untreated
horses demonstrated infections.
Since the 1950s the traditional insecticide treatments for control of
Hypoderma have been systemic organophosphates. These can kill migrat-
ing larvae but are relatively ineffective once the larvae are inside their
warbles. Trichlorphon (not in UK) and fenthion are among the most widely
used organophosphorus compounds at present (Boulard et al., 1991).
Unlike organophosphates, ivermectin is highly effective against all larval
instars and by the early 1980s was considered one of the most effective
systemics ever developed for use against Hypoderma (Scholl, 1993). Dis-
advantages of the use of ivermectin are the lengthy pre-slaughter with-
drawal period and that it should not be used in lactating animals
(Jackson, 1989).
Other macrocyclic lactones show similar effectiveness to ivermectin.
The milbemycin moxidectin (100400 pg kg-') was 100% effective
against migrating first instar H . lineatum (Scholl et al., 1992). Studies by
Hendrickx et al. (1993) have demonstrated that the avermectin, Dora-
mectin (200 pg kg-' subcutaneous) is 100% effective in the treatment of
cattle naturally infested with larvae of H . bovis at all stages of develop-
ment. In addition to the efficacy of Doramectin, it is notable that there were
no adverse reactions due to larval death as is sometimes noted when
organophosphates are applied. This is probably due to the mode of action
of avermectins, which cause a gradual paralysis of larvae followed by death,
rather than the sudden larval death with potentially massive release of
toxins (Eyre et al., 1981) following organophosphate treatments.
The pyrethroid Decamethrin was shown to be relatively ineffective as a
treatment for bovine hypodermosis when given either orally or as a pour-
on. In addition, high doses of 10 mg kg-' body weight caused severe
side effects of hyperactivity, perspiration and lachrymation (Boulard and

Troccon, 1984).
Tassi et al. (1987) have shown that subcutaneous injections of iver-
mectin at doses of 50, 100 and 200 pg kg-' were 100% effective against
goat warbles, Przhevalskiana silenus. The use of other insecticides for
controlLof this species is reviewed by Tassi et al. (1989).
Historically, control of Dermatobia hominis has been by the application
of various insecticides to the hide of livestock, to kill larvae in furuncles
and to prevent reinfestation. Applications had to be repeated at two- to
four-week intervals to be of benefit. Insecticides used in this manner
include toxaphene (camphechlor), DDT/gamma-BHC mixtures, crufo-
mate, fenthion and trichlorphon. The intramuscular injection of Closantel
(10-12.5 mg kg-') has been used for control with success, three injections
at regular intervals keeping cattle virtually free of larvae for three to four
months (Lancaster and Meisch, 1986). Decamethrine spray was up to 98%
effective as a curative treatment for Dermatobia hominis (Moriena et al.,
1984).
The methods used over the centuries in the control of D . hominis were
reviewed by Roncalli (1984a). Roncalli (1984a) also discussed the use of
ivermectin, which is the method of control for D. hominis most often
reported now in the research literature. Ivermectin is very effective for
killing larvae of D . hominis in cattle either, (1) by injection of IVOMEC@
Injection (at 200 pg kg-I body weight: Maia and Guimariies, 1986;
Roncalli and Usher, 1988; Moriena et al., 1984) or, (2) by topical applica-
tion of IVOMEC@Pour-On (at 0.5 mg kg-' body weight; Uribe et al.,
1989) or, (3) by sustained-release bolus (delivering 44-62 pg kg-' day-';
McMullin et al., 1989). All three treatments gave 95-99% reductions in
infestation rates. The injection and bolus had the advantage of remaining
active for about 90 days, whereas the topical application was effective for
about 30 days.
Abamectin (l%), an avermectin related to ivermectin, gave good treat-
ment against Dermatobia hominis when injected subcutaneously at a dose
rate of 200 pg kg-' (Cruz et al., 1993). Most larvae were expelled from the
hosts as a result of treatment, but a few shrunken, dead ones remained.
These were removed after ten days. What their effect would be if not
removed is unknown. Treated cattle remained free of D . hominis for 30
days, infestations only being detected again at the next examination, 44
days after treatment. Compared to untreated controls (4-29 larvae per
host), treated animals showed a marked improvement at 30 days, with
completely healed larval lesions, bright hair, weight gain and a generally
healthy appearance.
Another avermectin, Doramectin, has also recently been demonstrated to
have excellent therapeutic and prophylactic effect against Dermatobia
hominis in calves that were experimentally infested with first instar larvae
(Moya-Borja et al., 1993a). Thus, 48 h after treatment with a subcutaneous
injection of Doramectin _at a dose rate of 200 pg kg-', the number of
Dermatobia nodules was significantly reduced compared both to pre-
treatment levels and to controls treated with saline: there was 100% reduc-
tion in nodules after 6 days. At the same dose rate, Doramectin gave at least
35 days protection from infection by first instar larvae. This ability to
protect cattle from infestation gives chemicals such as Doramectin a
marked advantage over those that are only effective in treating existing
infections since, in economic terms, it is more important to prevent hide
damage than treat animals with an infection that has already damaged the
hide (Moya-Borja et al., 1993a). Dermatobia hominis is a seasonal pest,
* especially in the southern parts of its range, populations appearing in the
rainy season and disappearing in the cool, dry seasons. Strategic treatment
of cattle with Doramectin at regular intervals could provide protection from
Dermatobia throughout the season of fly activity.

Most of the control methods currently used aim to reduce host suscept-
ibility to myiasis either by killing eggs and feeding larvae with insecticides
or by reducing the number or suitability of available oviposition sites.
The current recommended treatment for wounds infested by Cochliomyia
hominivorax is a mixture of the organophosphates coumaphos (5%) and
chlorfenvinphos (2%) powder in a vegetable oil base. For Chrysomya
bezziana a range of insecticides have been shown to be effective
(Spradbery et al., 1991). A number of organophosphorus and synthetic
pyrethroid compounds are currently used in a variety of sprays, showers or
dips to control blowfly strike (Shanahan, 1965; Gruss, 1988; Gruss and van
Rensburg, 1988; French et al., 1994~).Ivermectin jetting fluid on sheep
gave over 93% protection against Lucilia cuprina strike 14 weeks after
treatment and could prove useful in regimens where it was used in rotation
with chemically unrelated insecticides to reduce resistance problems
(Eagleson et al., 1993).
Anthelmintics, such as ivermectin and orally administered Closantel
have also proved highly effective in the treatment of C . bezziana infesta-
tions (Spradbery et al., 1985; Spradbery and Owen, 1990; Reddy et al.,
1993) as has Doramectin for C. hominivorax (Moya-Borja et al., 1993b).
Most trials of insecticides for treatment of Wohlfahrtia myiasis have
been made in the former Soviet Union, where a wide range of organophos-
phorus compounds have been found to be effective, e.g. crotoxyphos,
trichlorphon, dichlorvos, propoxur, phosalone, temephos, chlorpyrifos,
iodofenphos and diazinon (Lancaster and Meisch, 1986). Work in
Kazakhstan demonstrated that treatment of infested wounds with 2%

emulsion or suspension of temephos, fenthion, propoxur and diazinon
was highly effective and gave protection for 5-7 days. For prophylaxis,
sheep should be sprayed within one day of shearing with an emulsion
containing 1% fenthion, propoxur or temephos or 0.1% diazinon at a
rate of one litre per sheep (Isimbekov and Zhanuzakov, 1983). Aerosol
applications of crotoxyphos and trichlorphon have also proven effective
(Simetskii, 1980; Podmogil’naya, 1983). These compounds have been
formulated as aerosol foams containing antibiotics, which form an elastic
waterproof film over the treated wounds.
.As a preventive treatment for the entire season, an emulsion of 0 . 1 4 2 %
diazinon has been sprayed every 3 4 days on all sheep at pasture from May
to November (Hadani et al., 1971). The incidence of Wohlfahrtia myiasis
was dramatically reduced and those cases that did occur were mild and
responded readily to curative treatment. However, this treatment regimen
represents a big investment in time and finance.
Treatment of W . magnijica infestation of camels in the Sinai has proven
effective with “Carcide”, a mixture of 2% diazinon, 0.05% pyrethrum and
0.25% piperonyl butoxide in pine oil (Hadani et al., 1989). The pine oil
appeared to repel gravid female flies.
Concerns over the development of insecticide resistance (e.g. see Wilson
and Heath, 1994), environmental contamination and effects on human and
animal health have led to work aimed at identifying alternatives to insecti-
cides (e.g. see Strong and Wall, 1990). This interest has resulted among
other things in numerous studies of hormonal disruption of insect growth.
Juvenile hormone, produced by the corpora allata in insects, plays a key
role in developmental processes such as larval moult, metamorphosis and
adult reproduction, including ovarian development. The juvenile hormones
and their synthetic mimics have long been known to have a range of
effects, including disruption of normal embryogenesis, moult inhibition,
interference with diapause, stimulation of precocious egg development
and, most commonly, the derangement of metamorphosis. Unfortunately,
juvenile hormones have, as yet, found little application against the higher
Diptera. However, other insect growth regulators (IGRs), such as the chitin
synthesis inhibitors, have been shown to be highly effective (Graf, 1993).
Of articular interest in this respect is the larvicide cyromazine (Vetra-
zinc!r, Ciba-Geigy). Cyromazine is a triazine compound which, when
applied as a pour-on may provide effective larvicidal cover against blow-
fly strike for up to 8 (Lonsdale et al., 1990) or even 13 weeks (O’Brien and
Fahey, 1991). It affects moulting and pupation following ingestion by first
instar larvae, but without affecting chitin synthesis (Friedel et al., 1988;
Graf, 1993). The development of systemic insect growth regulators, such as
lufenuron (Program@, Ciba-Geigy) a benzoylphenyl urea currently
available for flea control (Zakson et af., 1992), may also prove an impor-
tant future direction for myiasis control. Insect growth regulators are
particularly suited for use in strategic pest control and integrated pest
management but, since they do not act immediately and often do not kill
the pest at the stage where damage occurs, a better understanding of the
ecology and population dynamics of the pest is required for their effective
use.
6.2. Mechanical Means of Control
The reduction of conformational susceptibility to strike by Lucifia spp.,

‘through removal of skin folds and wool can also be brought about by
mechanical means. Dagging, the removal of faecally soiled wool, and
crutching, the regular shearing of wool from around the breech, can both
reduce susceptibility to strike by eliminating suitable oviposition sites.
Similarly, strike susceptibility is reduced in ewes following annual
shearing. Surgical removal of skin folds around the breech, the
“Mules” operation, is also used for Merino sheep in Australia (Gherardi,
1977; Townend, 1987). The scar tissue formed following this procedure
results in a smooth denuded area of skin, reducing faecal soiling and the
development of potential oviposition sites. However, the mulesing wound
can itself be attractive to L. cuprina during healing and the operation
should only be performed when blowflies are absent or in low abundance
(Cook and Steiner, 1990). Tail docking may also reduce the incidence of
strike in sheep (French et al., 1994b).
Hall and Smith (1993) review techniques for extracting larvae from
humans. Larvae of Cordyfobia anthropophaga can be expressed from
their furuncles relatively easily, either by pressure or by occlusion of the
opening with petroleum jelly, fats, water or similar substances which
prevent their respiration. Similar techniques have been traditionally used
by Indians of South America to extract Dermatobia hominis (Keech, 1981).
However, the morphology of second instar larvae of D . hominis, with an
anterior end broader than the diameter of the respiratory hole to the skin
surface, makes extraction more difficult, frequently necessitating surgery
(Lane et af., 1987). For removal of third instar Dermatobia, Nunzi et af.
(1984, 1986) first used surgery to enlarge the breathing pore and then
syringed anaesthetic (lidocaine hydrochloride; 2 ml/nodule) under the
larva to force it out of the furuncle by pressure. Li Loong et af. (1992)
describe the same extraction technique for Dermatobia and a similar
technique, with injection of hydrogen peroxide, has been used to extract
late second and third instar Hypoderma from the backs of cattle (Scholl and
Barrett, 1986).
MWASIS OF HUMANS AND DOMESTIC ANIMALS 299
6.3. Biological Control
Kal’vish (1990) reported that application of a suspension of spores of the

entomopathogenic fungus Tolypocladiurn niveurn to adult Oestrus ovis
caused 100% mortality (compared to 30% of controls). Such deleterious
relationships between insects and fungi have been comprehensively
reviewed by Hajek and St Leger (1994).
Parasites, pathogens or predators may play a more important role than
previously thought in limiting the distribution of Hypoderma in the
northern hemisphere (Scholl, 1990) and studies of them could be of value
in indicating future biological control agents.
The effects of phoretic mites, Macrocheles rnuscaedornesticae, on
Dermatobia horninis in the laboratory was to reduce their insemination
rate (from 75% to 17%), probably by mechanical interference with copula-
tion, and to reduce the ability of females to capture and oviposit on egg-
carriers, Musca dornestica. The longevity and fertility of the Dermatobia
were not affected (Moya-Borja, 198 1). Macrocheles rnuscaedornesticae
have also been found infesting Cochliornyia hominivorax in Libya,
together with the parasitic mite, Trichotrornidiurn muscarurn, but their
effects on adult screwworm are unknown (McGarry et al., 1992).
Wettable powder formulations of Bacillus thuringiensis var wuhhanensis
have been used in small-scale field tests on sheep for the prevention of
artificially induced strike by Lucilia cuprina (Cooper et al., 1985). Persis-
tent biological activity and control were effected for 21 days, suggesting
that B. thuringiensis may be of value as a future biological control agent.
6.4. Sterile Insect Technique
The release of irradiated sterile males into a wild population is known as

the sterile insect technique (SIT) (Knipling, 1955). Females mating with
irradiated males produce eggs that fail to hatch. If enough irradiated males
are released and obtain a large proportion of the matings with fertile
females, with continued release of sterile males the population of wild
insects is eventually driven to extinction.
6.4.1. Oestridae
The major problem with application of SIT to oestrid flies is the extreme
difficulty of rearing the species. Beesley (1967) cultured first instar larvae
of Hypoderrna in vitro: a few moulted to second instar but did not develop
further. Techniques were described more recently for enhanced survival of
H. lineaturn in artificial media (Chamberlain and Scholl, 1991), but these
techniques only applied to third instar larvae removed from cattle at

slaughter. Despite the difficulties in producing sterile Hypoderma, release
of sterile males of H . bovis and H . lineatum was successful in eliminating
these species in a small trial along the USA-Canada border (Kunz et al.,
1990), following earlier insecticidal applications to cattle to reduce the
wild populations to a low level. This integrated approach is an attractive
option for the future, because the combined treatment attacked both
immature and adult stages of the parasites in a complementary manner.
However, before large-scale integrated control programmes using SIT can
be initiated, in vitro rearing techniques will need to be developed.
Although sterile Dermatobia hominis can:be produced by gamma radia-
.tion and by application of chemosterilants (Moya-Boija and Borkovec,
1981), no sterile fly control activities have been begun because attempts
to culture D. hominis in vitro have so far been only partly satisfactory
(Zeled6n and Silva, 1987). First instar larvae grew well on a mixture of
50% Eagle’s MEM,40% fresh bovine serum and 10% yeast extract, with
antibiotics (penicillin, streptomycin, nystatin), but only 29% moulted to the
second instar and thereafter development was very poor, with only 0.7%
moulting to third instar at which they died. More success was had with
rearing second and third instar larvae removed from cattle, but results were
still unsatisfactory: only 39% of second instars moulted to third instar and,
of a small subsample of third instars that pupated, only 16% hatched
(Zeledbn and Silva, 1987).
6.4.2. Calliphoridae
The use of SIT against the screwworm fly Cochliomyia hominivorax in
North and Central America, supported by insecticidal treatments of live-
stock, restrictions on movement of infected animals and an intensive
idormation campaign, has been documented in detail (Graham, 1985;
Krafsur et al., 1987). The New World screwworm is ideally suited to
mass rearing techniques, nowadays achieved using a gelled diet (Taylor,
1992). Mass production is aided by selection for laboratory traits of
increased fecundity and shortened development time, but these must be
balanced against selection for undesirable traits that reduce effectiveness of
the strain on release (Thomas, 1993b).
The inadvertent introduction of C. hominivorax screwworm flies in
North Africa in 1988 led to the implementation of an international pro-
gramme to eradicate the fly using SIT, co-ordinated by the Food and
Agriculture asanisation of the United Nations (Lindquist et al., 1992).
Cochliomyia hominivorax were reared at the screwworm production facil-
ity of the Mexican-American Commission for Eradication of Screw-worm
in Tuxtla Gutidrrez, Mexico. They were sterilized by exposure to gamma
MWASIS OF HUMANS AND DOMESTIC ANIMALS 30 1
radiation as late-stage pupae, then placed in small cardboard boxes (1600

pupae per box) together with a sugar-based food supply to sustain the
emerging flies. The boxed pupae were then transported to Libya, in the
final phases direct from Tuxtla to Tripoli by charter jet. When approxi-
mately 80% of the adults had emerged, the boxes were dispersed at about
three to ten boxes per minute, from light aircraft flying at 240 km h-' along
predetermined flight paths about 4 km apart. The boxes opened in mid air or
on impact with the ground releasing the newly emerged sterile adults.
Full-scale releases began in February 1991, at the time when the screw-
worm fly population was at its seasonal minimum. Flies were released at
initial rates of 3.5 million per week rising to peak rates in July 1991 of 40
million per week, at densities ranging from 500-1200 km-'. By May 1991
an area of 41 000 km2 was being treated each week. The release of sterile
males was backed-up by extensive control operations on the ground,
including surveillance teams and quarantine stations to prevent movement
of infested animals outside the infested area. In 1990 a total of 12 068
confirmed cases of C. hominivorax infestation had been recorded, but in
1991 only six cases were seen, the last occurring in April. By November,
after six months with no further cases, the release of sterile flies was
terminated. The official announcement of the success of the eradication
campaign was made on 22 June 1992, and it has been estimated that the
benefit:cost ratio of the campaign for the North African region was 50:l
(Cunningham et al., 1992; FAO, 1992). A technique developed for dis-
persing sterile Chrysomya bezziana (Spradbery et al., 1989) is now being
used with success in Central America for dispersal of sterile C. homini-
vorux. In the Chilled Fly Dispersal System (CFDS), newly emerged flies
are inactivated by chilling and can then be transported in bulk in tempera-
ture regulated containers that can be fitted into light aircraft, so avoiding
the direct and indirect expenses of boxed pupae. The flies are reactivated
by warming when they contact the outside air as they are released during
dispersal flights (M. Vargas-Terh, personal communication).
Preliminary field trials, using sterile male Chrysomya bezziana in Papua
New Guinea have also indicated the potential value of SIT for the control
of this species (Spradbery et al., 1989). Given the threat to the Australian
livestock industry posed by the inadvertent introduction of C. bezziana,
the ready availability of mass-reared flies and SIT technology may be
particularly important (Spradbery, 1994).
6.5. Genetic Control
Exposure of Lucilia cuprina to radiation was found to induce a range of

mutations, including translocations, where one or more sections of the Y
chromosome are swapped with sections of one or more autosomes. Males

possessing the translocation can be identified and selected in the laboratory.
Possession of the Y-linked translocations confers partial sterility and a
proportion of the eggs produced in a cross between a modified male and
a wild female will fail to hatch. Among the individuals that do hatch the
deleterious translocation is carried by the male progeny but not the females
(Whitten ef af., 1977; Whitten, 1979).
Further work has shown that it is also possible to translocate onto the Y
chromosome the wild-type alleles of autosomal mutations affecting eye
colour. The recessive mutations are carried on the non-translocation
chromosome set, by the genetically modified males and in heterozygotic
. females. However, they are not expressed because of the presence of the
dominant wild-type alleles. In female homozygotes, the mutations are
expressed as white or yellow eye colour. These females lack the light-
filtering pigments that give the normal blowfly eye its reddish-brown
colour, so the homozygous mutant flies are effectively blind in daylight.
When heterozygous females mate with the genetically altered males a large
percentage of their female progeny are homozygous for one or more of the
eye pigment mutations. The blind females can be reared in the laboratory but
die rapidly in the field. The system is termed a female-killing (FX) system or
the genetically impaired female technique (GIFT) (Foster et af., 1992).
Release of the modified strains causes genetic death, partly from semi-
sterility caused by the chromosome rearrangement and partly from death of
the female descendants of the released males due to homozygosity for the
mutations. An additional development is that the released males also
carry inversions to prevent recombination and maintain the linkage of
the deleterious genes to the translocation. The inversion also contributes
an additional degree of potential sterility for females which inherit the
inversions in heterozygous form.
Computer simulations indicated that a theoretical maximum death rate of
98% per generation could be achieved by release of males possessing the
chromosome abnormalities and the eye colour mutations (Foster et al.,
1988). Furthermore, genetic death from semisterility and homozygosity
should persist in the wild population for several generations after cessation
of releases, giving this control system a considerable advantage over
conventional SIT in which suppression ceases when release stops. At
lower release rates GIFT would be expected to result in more rapid
suppression of the target population than SIT.
In field trials on Flinders Island, which has an area of about 40 km2and
is 27 km off the south Australian coast, 34 000 modified male L. cuprina
were released per km2each week between August 1985 and May 1986. The
induced rate of genetic death peaked at 87%, 6 months after the trial began
and the blowfly population declined from 345 females per hectare in
October 1985 to less than one female per hectare in May 1986, when
releases were terminated (Foster and Smith, 1991). The population
remained at below four females per hectare for the following 10 months
(Foster, 1989; R. Mahon, personal communication).
However, although the Hinders Island trial showed that it is possible to
reduce an isolated Lucilia population by this method, further trials in the
much larger Furneaux Island group, encountered significant problems.
Difficulties were experienced in rearing the 15 million modified flies
required per week to swamp the wild population and problems arose
from recombination within males carrying the translocation under large-
ssale rearing conditions. As a result of these considerations, together with
the expense of maintaining large-scale rearing facilities, removal of L.
cuprina from large areas of the Australian mainland using this technique
has not so far been attempted.
6.6. Vaccines
Much work has been carried out to characterize bovine immune responses
to Hypoderma infestations, reviewed by Baron and Weintraub (1987),
Baron and Colwell (1991b) and Scholl (1993). This work is based on the
observation that, generally, fewer larvae appear in the back of older cattle
than in calves or yearlings, implying that some immunity develops with
age. The principal antigens appear to be Hypoderma digestive enzymes and
four proteinase fractions have been identified: hypodermin A, hypodermin
B and hypodennin C (collagenase) and P2. Recent work by Baron and
Colwell (1991a) involved a comparison of calves immunized with a
purified combination of hypodermins A, B and C, and monophosphoryl
lipid A (MPL, a potent immune system stimulant), calves treated with MPL
alone and untreated control calves. All calves were given a subcutaneous
injection of 100 first instar larvae of H. lineatum one week after a two-week
immunization schedule. The maximum number of grubs appearing in the
back of the antigen+MPL group was significantly lower than that of the
other two groups, but the mean number of viable larvae (after death of
grubs in the back) was not different between the two treated groups,
although both were significantly lower than in the control group. MPL
enhanced antigen specific lymphocyte responsiveness in immunized calves
by completion of the immunization schedule. It also enhanced lymphocyte
responsiveness to antigen stimulation in calves receiving only MPL by four
weeks post-infection. Baron and Colwell (1991a) concluded that stimula-
tion of a strong antigen-specific cellular immune response prior to infesta-
tion could significantly reduce the survival of H. lineatum, but the role of
antibodies in that response remains to be determined. There is no apparent
correlation between the development of humoral antibodies and resistance

to Hypoderma infestations (Gingrich, 1982; Baron and Weintraub, 1987).
Chabaudie et al. (1991) vaccinated cattle with hypodermin A only, with or
without adjuvants. They did not demonstrate any significant difference
between vaccinated and control groups of calves, in the number of Hypo-
derma infestations naturally acquired when the calves were released onto
pasture. These results again suggest that humoral immunity is not involved
in acquired resistance to reinfestation by Hypoderma: the factors which are
involved are still unknown (Chabaudie et al., 1991).
Current research is examining the causes of immunosuppression noted
during infestations with Hypoderma, when the absence of any inflam-
matory reaction around the first instar larvae indicates that these larvae
may avoid the host’s non-specific defence mechanisms. Hypodermin A is
a powerful anti-inflammatory factor and appears to play a crucial role in
mediating the immunosuppression reported in bovine hypodermosis
(Chabaudie and Boulard, 1992a, b); conversely, hypodermin C had no
such effect (Chabaudie and Boulard, 1993).
Discovery of the host humoral response to antigens of Lucilia cuprina
has stimulated considerable interest in the production of a vaccine.
Vaccination of sheep with a partially purified extract of L. cuprina larvae
can result in a marked reduction in growth when larvae are fed on sheep
(Johnston et al., 1992). Experimental vaccines have been produced based
on serine proteases secreted by larvae (Sandeman, 1990) and larval mem-
brane proteins (Johnston et al., 1992), with effects being amplified by
adjuvants (East et al., 1992). However, to date, no effective commercially
available vaccine has been developed. An effective approach to the pro-
duction of vaccines against blowfly larvae would require a readily available
source of purified antigen. To date, in vitro culture has not been possible
(Baron and Colwell, 1991b).
A novel approach to the search for vaccines may be to target antigens
that do not normally stimulate an immune response. Thus, antigens in the
cuticle (Barrett and Trevella, 1989) and parasite gut could be targeted:
initial studies have shown that first instar larvae of L. cuprina, feeding on
sera from animals vaccinated with a protein found attached to the peri-
trophic membrane of the larval gut, have their growth decreased by up to
60% (Willadsen et al., 1993). Eisemann et al. (1993) studied the uptake of
antibody by feeding larvae of L. cuprina and demonstrated that there was a
progressive decrease in the percentage of ingested antibody remaining
functional, from 90% in the crop to around 10% in the posterior midgut
and hindgut, presumably due to the action of proteolytic enzymes on the
antibodies during digestion. Therefore, the anterior half of the gut and its
contents offer the best prospects as targets for vaccination, although the
foregut and midgut epithelial cells may be protected from antibodies by,
respectively, a cuticle layer and the peritrophic membrane.
6.7. Baits, Traps and Targets
6.7.1. Trapping for Control

A screwworm adult suppression system (SWASS) developed for the New
World screwworm fly Cochliomyia hominivorux uses a chemical lure, food
and 2% dichlorvos to attract and kill adult flies (Coppedge et al., 1978a).
The synthetic chemical attractant is known as swormlure (Coppedge et al.,
1977; Mackley and Brown, 1984). The bait was developed from analysis of
the volatiles released from decomposing blood and carrion (Grabbe and
Turner, 1973) and promising mixtures tested in the field (Jones et al., 1976;
Coppedge et al., 1977). The current formulation is composed of a mixture
of butyl alcohol, dimethyl disulphide, phenol, 2-methylphenol, indole, and
ethanoic, butanoic, pentanoic, and benzoic acids (Mackley and Brown,
1984). Swormlure increases the catch of C . hominivorux while decreasing
the non-target fly catch compared to liver baits (Coppedge et al., 1977).
Field trials with the SWASS gave a 6 5 8 5 % reduction in an isolated wild
C . hominivorux population within three months (Coppedge et al., 1978b).
However, environmental concerns about the release of large quantities of
dichlorvos has resulted in the SWASS being largely abandoned as a control
technique.
In field trials, the incidence of strike by Lucilia cuprina in Australia was
reduced by over 50% using carrion-baited traps (Mackerras et al., 1936).
However, at that time the use of traps for blowfly control was not con-
sidered economic due to high construction costs, poor efficiency, the large
number of traps needed and the frequency of bait-replacement and trap
maintenance required. More recently a number of studies have renewed
interest in the development of simple catching systems for blowfly popula-
tion suppression. Work in Australia has used bait-bins filled with carrion
and insecticide to attract and kill L. cuprina. Preliminary data have
suggested that bait-bins may be able to reduce fly abundance and the
incidence of flystrike (Anderson, J.M.E. et al., 1990). Similarly, current
work in Britain is developing coloured targets which could be used with an
insecticide, IGR or chemosterilant to attract and kill or sterilize L. sericata
(Wall et al., 1992b), particularly if accompanied by a powerful synthetic
odour bait (Ashworth and Wall, 1994). No field trials of the efficacy of the
target system have yet been undertaken. Related work is developing targets
for Cochliomyia hominivorax (Torr and Hall, 1992; Green et al., 1993) and
Wohlfahrtia magnifica (Hall et al., 1995).
7. NEW MONITORING, MODELLING AND FORECASTING METHODS
7.1. Monitoring by Serodiagnosis
Diagnosis of Oestrus ovis infestation in sheep can be problematic because

symptoms can be confused with other respiratory diseases and because
sheep with overwintering larvae (usually first instar) do not show any
clinical signs. Bautista-Garfias et al. (1988) investigated the use of sero-
logical techniques for diagnosis, using double immunodiffusion (DD) and
indirect haemagglutination (IH) tests with somatic crude antigens prepared
from first, second and third instar larvae. IH tests were most sensitive,
*
especially using first or second instar antigens: however, IH tests using

third instar antigens gave many false positives with uninfested sheep,
indicating that more studies on cross-immunity to other parasites of sheep
are needed. Jagannath et al. (1989~)also observed that second instar larvae
were more immunogenic than third instar larvae by IH, but the false
negatives and potentially false positives in their work sound a cautionary
note regarding the interpretation of serodiagnostic results. The problems
of cross-reactions in ELISA-based serodiagnosis were addressed by
Haralampidis (1987) who absorbed out non-specific immunoglobulins.
As with Oestrus, serological techniques can be used for the diagnosis of
Gasterophilus. Ribbeck et al. (1984) used counterimmunoelectrophoresis,
passive haemagglutination and intradermal injection to test for Gastero-
philus infestation, using antigen prepared from larvae of six species: all
three techniques gave positive results. Escartin-Peiia and Bautista-Garfias
(1993) demonstrated the effectiveness of thin layer immunoassay, diffu-
sion-in-gel ELISA and, to a lesser extent, indirect haemagglutination for
.detection of antibodies in horse and donkey sera reacting to antigen
prepared from third instar G. intestinalis. As with Oestrus ovis, false
positives were recorded, possibly due to cross-reactions with antibodies
to other parasites or possibly due to previous infections. Further work is
needed to improve the sensitivity and selectivity of such serodiagnosis,
possibly using antigen prepared from earlier instar larvae as for Oestrus
ovis. The larval proteins with antigenic activity differ between larval
instars (Ilchmann et al., 1989).
The use of ELISA for detection of hypodermosis in cattle (Sinclair and
Wassall, 1983) is now well accepted (Tarry, 1986; Colwell and Baron,
1990), and preliminary evaluation of immunological diagnosis of Hypo-
derma tarandi in reindeer looks promising (Monfray and Boulard, 1990).
To reduce costs in large epidemiological surveys for hypodermosis, analysis
of pooled serum and milk samples (from 10 hosts) has been successfully
tested (Boulard and Villejoubert, 1991). A model for estimating the
abundance of Hypoderma from the proportion of uninfested calves in a

herd, as determined by serological techniques, has been developed by Lysk
et al. (1991). This model should be of value in routine monitoring, asses-
sing control programmes and in determining release rates for sterile insect
programmes.
Boulard and Petithory (1977) first demonstrated that human hypo-
dermosis can be diagnosed serologically by immunoelectrophoresis, using
proteolytic secretions of the first instar larvae as antigens. The ELISA and,
to a lesser extent, passive haemagglutination tests have subsequently been
shown to be more sensitive, but immunoelectrophoresis is better able to
track the temporal parasitological progress of the disease (Doby et al.,
1987).
Recent work has indicated the potential value of serological techniques
to indirectly investigate prevalence of myiasis due to Cochliomyia homini-
vorax. Thomas and Pruett (1989) noted that antibodies to C. hominivorux
antigens were found in the serum of previously infested sheep for at least a
month after infestation. However, the present antigens need refinement to
prevent cross-reactivity giving false positives to previous infestations with
other myiasis-producing calliphorids (Thomas and Pruett, 1992).
7.2 Monitoring by Traps
A wind-oriented trap, which employs a swormlure bait, is used to monitor

populations of Cochliomyia hominivorux (Broce et al., 1977). Swormlure
is also used as an attractant in sticky traps which were developed for
sampling populations of Chrysomya bezziana (Spradbery, 1981). These
traps are widely used in monitoring the progress of the C. hominivorux
eradication programme in central America and for screwworm fly surveil-
lance in northern Australia. However, the importance of vision and colour
in the response of C. hominivorax to targets has recently been demonstrated
in the laboratory (Green and Warnes, 1992) and targets with a greater
visual component than the wind-oriented trap were shown to be signifi-
cantly more attractive to C. hominivorax in the field (Ton and Hall, 1992).
Furthermore, swormlure appears to act primarily as a feeding attractant,
attracts fewer gravid females and is still a relatively weak attractant
compared to a wounded host animal (Thomas and Mangan, 1989).
Hence, there appears to be considerable potential for the further enhance-
ment of the visual attractiveness and odour composition of the baits used in
monitoring systems for these species. Linked gas chromatography and
electroantennography has recently been used to identify compounds in
larval wound fluid that stimulate activity in the antenna1 receptors of C.
hominivorax and that might, therefore, be important in wound location
(Cork, 1994). Such work might lead to new odour baits simulating wound
odours that attract a greater proportion of gravid females than swormlure.
Carrion-baited traps (Vogt and Havenstein, 1974; Williams, 1977) and
sticky traps (Wardhaugh et al., 1984) have been used for monitoring
populations of Lucilia cuprina and coloured, sticky targets for L. sericata
(Wall et al., 1992b, 1993a). Catches made by baited traps and targets are
however, subject to a considerable range of biases as a result of weather,
sex and stage in the reproductive cycle at which flies are more or less
attracted. All of these need to be considered in the interpretation of catch
data (Vogt et al., 1983, 1985b; Vogt and Morton, 1991; Spradbery and
Vogt, 1993; Wall, 1993).
7.3. Modelling and Forecasting
Predictive models may be of value in helping to assess the likely threats of

the geographic spread of specific fly species and their economic con-
sequences. To this end a climate matching programme (CLIMEX) has
been developed which indicates the potential geographic distribution and
relative abundance of a species. The model has been used to show that
Australia and the Americas are particularly at risk from invasion by
Chrysomya bezziana, the potential global distribution of which is similar
to that recorded for Cochliomyia hominivorux (Sutherst et al., 1989). The
model predicted that C.bezziana could occur in Ethiopia beyond its then
known limits, in Africa. This prediction was accurate because C. bezziana
has since been recorded from Gondar Province, Ethiopia (A. Demissie and
M. Hall, unpublished).
In the control of myiasis and/or suppression of adult fly populations,
effectiveness may be greatly enhanced by appropriate timing of control
measures within the pattern of seasonal incidence or abundance (McKenzie
and Anderson, 1990). However, for the operation of such a policy, it is
necessary to be able to predict the forthcoming changes in myiasis inci-
dence or fly abundance, at least in the short term. The development of such
predictive models would allow farmers to use insecticides prophylactically
and may even allow less radical alternative techniques, such as trap/target
population suppression systems to be employed cost effectively.
One such model is currently being developed to predict the seasonal
changes in fly abundance and sheep-strike incidence by Lucilia sericata.
Using an analysis of the day-degree requirements of each L. sericata life-
cycle stage and measurement of daily temperatures, the model developed to
date is able to explain a high proportion of the variance in fly catch and can
predict relative fly abundance, given appropriate temperature measure-
ments, extrapolations or forecasts (Wall et al., 1992c, 1993a). The model
has been used to explore the effects of a range of control options on L.

sericata abundance. The optimum control policy for fly population sup-
pression was shown to be strategic early spring treatment of stock, to
ensure that a high proportion of the offspring of the first generation of
the season are eliminated (Wall et al., 1993b). The effectiveness of spring
strategic treatment has been tested in the field with promising results.
Further work is underway to determine the relationships between fly
abundance, strike incidence and other factors such as weather in this
system (French er al., 1994a).
Working towards a similar goal, but from the opposite perspective,
Wardhaugh and Morton ( 1990) have developed predictive equations
which show that, in Australia, 76% of the observed variance in body
strike and 54% of that for crutch strike may be explained by measurement
of a combination of weather variables, indices of pasture growth and the
population density and activity of gravid L. cuprina. They are also now
working to complete their understanding of the causes of changes in fly
abundance and strike epidemiology so that they can develop a compre-
hensive predictive model for use in the Australian pastoral system.
8. CONCLUSIONS AND FUTURE
When James (1947) wrote his important work on myiasis of humans he

observed that there had been little quantitative work done on myiasis: at that
time it was a rather descriptive, case-oriented branch of medical and
veterinary entomology. Since then, analytical studies of myiasis have
increased dramatically, particularly in alliance with new control and moni-
toring methods that demand an increased understanding of the biology,
physiology and ecology of the parasites, for example, SIT, serodiagnosis,
vaccine research and trapping.
Despite the increased understanding of myiasis, as reported here, there
are still many aspects about which we know relatively little. The distribu-
tions and economic impacts of even the important species are poorly
recorded. The incidence of myiasis in wildlife is similarly not well
recorded, nor is the role of wildlife as reservoirs of infection for humans
and domestic animals. Serodiagnosis could usefully be applied to wildlife
to survey for levels of infection on a wider scale than has previously been
done (Thomas and Pruett, 1989).
Greater thought needs to be given to the development of coherent
myiasis control policies, and the relevant merits of eradication or popula-
tion suppression and disease management. To enhance possibilities of
developing such policies, trapping for monitoring and control is an area
that has dttracted increasing recent attention. However, most of the work
has beeg conducted on calliphorids and sarcophagids. This could usefully
be extended to further work on the behaviour of Oestridae, as shown by the
detailed studies of reindeer bot flies, Cephenemyia trompe, at reindeer and
reindeer-like traps (Anderson, 1975, 1989; Anderson and Nilssen, 1990).
It is clear that avermectins have had a dramatic impact on control of
myiasis species, oestrids in particular. However, the use of avermectins for
prophylactic control of myiasis will necessitate the maintenance of high
blood concentrations over long periods, administered by multiple treat-
ments or by rumenal bolus. Avermectins eliminated in cattle dung have
been shown to have a significant impact on non-target dung fauna, parti-
cularly beetle larvae, since a high proportion of the administered dose is
eliminated in the faeces where it may remain toxic (Strong, 1992). Con-
sequently such prophylaxis is likely to be environmentally undesirable,
although the debate is ongoing (Herd et al., 1993).
The recent introduction of Cochliomyia hominivorax into Libya, followed
by its costly eradication (US$75 million: Cunningham et al., 1992),
indicates the real need for more detailed information on the behaviour,
ecology and life cycles of myiasis species. Such information is important
not only for control programmes in the endemic areas but can provide
predictions on species performance in areas into which they might be
introduced as exotic pests. At its time of introduction to Libya, C . hornini-
vorax was not a notifiable pest and there were no international regulations
regarding the transport of infected livestock. It is now included on Disease
List B of the Office International des Epizooties (FAO, 1992). However,
myiasis due to Chrysomya bezziana is not included on the list despite the
facts that it has the potential to be as serious a pest as C. hominivorax
(Spradbery, 1994) and has previously been redistributed through countries
of the Persian Gulf with livestock (Rajapaksa and Spradbery, 1989). It is
worth noting that accidental transfer of screwworms in infested hosts is a
risk not just with animal vectors but with humans too (Mehr et al., 1991).
The current threat of reintroduction of Hypoderma to the UK is a further
example of the dangers of passage of myiasis species with infested live-
stock (Tarry, 1994). However, it should be remembered that such move-
ments of myiasis species are not a new phenomenon: Hypoderma,
Gasterophilus, Oestrus and Lucilia have all had their ranges greatly
extended by human activities.
There is considerable scope for further study of the interactions between
agents of myiasis and other parasites of domestic livestock, regarding
immunosuppression and the possibility of myiasis making animals more
susceptible to other diseases. Similarly, there is scope for further integra-
tion of monitoring and control programmes for myiasis species with those
for other diseases, for example, the monitoring for hypodermosis in the UK
using sera collected for brucellosis monitoring (Tarry, 1986). A further

area for future study is the difference between strains of species in their
virulence and, for facultative species, their prevalence as parasites. Thus,
Lucilia sericrrfu is a serious pest in Europe but appears not to be so in North
America, whereas the opposite situation holds for Phormiu regina (James,
1947; Zumpt, 1965). Likewise, L. cuprina is a major pest in Australia but
not in North America. The reasons for such differences have not been
studied. Problems of intraspecific variation and its implications for pest
management are considered in some detail by Kim and McPherson (1993),
with particular reference to crop pests.
:In conclusion, many recent contributions have been made to our under-
standing of myiasis, its agents and their interactions with host animals, but
much remains to be done. It is hoped that this review will be a positive
stimulus for future activities in this field.
ACKNOWLEDGEMENTS
We are very grateful to all those who answered numerous requests for
reprints and other information on myiasis and to the staff of the Ento-
mology Library, Natural History Museum, for their great help in tracing
references. We are also grateful to Keith Wardhaugh, David Tarry and Paul
Ready for their comments on an earlier draft of the manuscript.
REFERENCES
Alexander, J. O’D. (1984). “Arthropods and Human Skin”. Springer-Verlag,

Berlin, 422 pp.
Amarante, A.F.T., Barbosa, M.A., Oliveira-Sequeira, T.C. and Femandes, S.
(1992). Epidemiology of sheep myiases in Sao Paulo State, Brazil. Tropical
Animal Health and Production 24, 36-39.
A m , Z.S., Amr,B.A. and Abo-Shehada, M.N. (1993). Ophthalmomyiasis externa
caused by Oestrus ovis L. in the Ajloun area of northern Jordan. Annals of
Tropical Medicine and Parasitology 87, 259-262.
Anderson, J.M.E., Shipp, E. and Anderson, P.J. (1984). Blowfly populations and
strike incidence in the arid zone of N.S.W. Wool Technology and Sheep
Breeding 32, 12-14.
Anderson, J.M.E., McLeod, L.J., Shipp, E., Swan, A. and Kennedy, J.P. (1990).
Trapping sheep blowflies using bait-bins. Australian Veterinary Journal 67,
93-97.
Anderson, J.R. (1975). The behaviour of nose bot flies (Cephenemyia apicata
and C . jellisoni) when attacking black-tailed deer (Odocoileus hemionus
columbianus) and the resulting reactions of the deer. Canadian Journal of

Zoology 53,977-992.
Anderson, J.R. (1989). Use of deer models to study larviposition by wild naso-
pharyngeal bot flies (Diptera: Oestridae). Journal of Medical Entomology 26,
234-236.
Anderson, J.R. and Nilssen, A.C. (1990). The method by which Cephenemyia
trornpe (Modeer) larvae invade reindeer (Rangifer tarandus). Rangifer Special
Issue No. 3, 291-297.
Anderson, P.J.E., Shipp, E., Anderson, J.M.E. and Dobbie, W. (1988). Population
maintenance of Lucilia cuprina (Weidemann) in the Arid Zone. Australian
Journal of Zoology 36, 241-249.
Anderson, H. (1988). De svenska styngen. Entomologisk Tidskrift 109, 3 1 4 1 .
Arbit, E., Varon, R.E. and Brem, S.S. (1986). Myiatic scalp and skull infection
. with Diptera Sarcophaga: case report. Neurosurgery 18, 361-362.
Arm, E., Puccini, V., Schindler, P. and Tassi, P. (1982). Efficacia del rafoxanide
iniettabile e orale contro le larve de Oestrus ovis in ovini infettati naturalmente.
Atti della Societa Italiana delle Scienze Veterinarie 36, 662-664.
Ashworth, J.R. and Wall, R. (1994). Responses of the sheep blowflies Lucilia
sericata and L. cuprina to odour and the development of semiochemical baits.
Medical and Veterinary Entomology 8, 303-309.
Aubertin, D. (1933). Revision of the genus Lucilia R.-D. (Diptera, Calliphoridae).
Journal of the Linnean Society 33, 389436.
Baily, G.G. and Moody, A.H. (1985). Cutaneous myiasis caused by larvae of
Cordylobia anthropophaga acquired in Europe. British Medical Journal 290,
1473-1474.
Baird, J.K., Baird, C.R. and Sabrosky, C.W. (1989). North American cuterebrid
myiasis: report of seventeen new infections of human beings and review of the
disease. Journal of the American Academy of Dermatology 21, 763-772.
Barnard, D.R. (1977). Skeletal-muscular mechanisms of the larva of Lucilia
sericata (Meigen) in relation to feeding habit. Pan-Pacific Entomologist 53,
223-229.
Baron, R.W. and Colwell, D.D. (1991a). Enhanced resistance to cattle grub
infestation (Hypoderma lineatum de Vill.) in calves immunized with purified
hypodermin A, B and C plus monophosphoryl lipid A (MPL). Veterinary
Baron, R.W. and Colwell, D.D. (1991b). Mammalian immune responses to myia-
sis. Parasitology Today 7,353-355.
Baron, R.W. and Weintraub, J. (1987). Immunological responses to parasitic
' arthropods. Parasitology Today 3, 77-82.
Barrett, M. and Trevella, W. (1989). The immune response of the sheep popliteal
lymph node to a purified phenoloxidase from larval cuticle of the sheep
ectoparasite, Lucilia cuprina. Journal of Parasitology 75, 70-75.
Barton Browne, L., Van Gerwen, A.C.M. and Williams, K.L. (1979). Oocyte
resorption during ovarian development in the blowfly Lucilia cuprina. Journal
of Insect Physiology 25, 147-153.
Baruch, E., Godel, V., Lazar M., Gold, D. and Lengy, J. (1982). Severe external
ophthalmomyiasis due to larvae of Wohlfahrtia sp. Israel Journal of Medical
Sciences 18, 8 15-8 16.
Baudet, E.A.R.F. and Nieschultz, 0. (1938). Uber Fliegenlarveninfektion (Lucilia
seriata) bei Schafen im Wieringermeerpolder (Zuiderseegebiet). Zeitschrift f i r
Angewandte Entomologie 20, 324-325.
Baumgartner, D.L. (1988). Review of myiasis (Insecta: Diptera: Calliphoridae,

Sarcophagidae) of Nearctic wildlife. Wildlife Rehabilitation 7 , 3 4 6 .
Baumgartner, D.L. (1993). Review of Chrysomya rujifaces (Diptera: Calli-
phoridae). Journal of Medical Entomology 30,338-352.
Baumgartner, D.L. and Greenberg, G. (1984). The genus Chrysomya (Diptera:
Calliphoridae) in the New World. Journal of Medical Entomology 21, 105-1 13.
Bautista-Garfias, C.R., Angulo-Contreras, R.M. and Garay-Garzon, E. (1988).
Serological diagnosis of Oestrus ovis (Diptera: Oestridae) in naturally infested
sheep. Medical and Veterinary Entomology 2, 35 1-355.
Beamer, R.H. (1950). An observation on the egg-laying of Cuterebra buccata Fabr.
in nature. Journal of the Kansas Entomological Society 23, 16.
Beamer, R.H., Penner, L.R. and Hibbard, C.W. (1943). Some notes on the biology
.of the pack rat cuterebrid (Cuterebrid beameri Hall) in Kansas. Journal of the
Kansas Entomological Society 16,47-50.
Beck, T., Moir, B. and Meppen, T. (1985). The cost of parasites to the Australian
sheep industry. Quarterly Review of Rural Economics 7 , 336-343.
Beesley, W.N. (1967). Observations on the biology of the ox warble-fly (Hypo-
derma:'Diptera, Oestridae). I.-ln vitro culture of the first-instar larva. Annals of
Tropical Medicine and Parasitology 61, 175-1 8 1.
Bennett, G.F. and Whitworth, T.L. (1991). Studies on the life history of some
species of Protocalliphora (Diptera: Calliphoridae). Canadian Journal of Zool-
ogy 68,2048-2058.
Blacklock, B. and Thompson, M.G. (1923). A study of the tumbu-fly, Cordylobia
anthropophaga Grilnberg, in Sierra Leone. Annals of Tropical Medicine and
Parasitology 17,443-502.
Boulard, C. and Petithory, J. (1977). Serological diagnosis of human hypodermo-
sis: a preliminary report. Veterinary Parasitology 3,259-263.
Boulard, C. and Troccon, J.L. (1984). Effect of dose and route of application on the
action of Decamethrin against cattle grubs. Veterinary Parasitology 14,65-74.
Boulard, C. and Villejoubert, C. (1991). Use of pooled serum or milk samples
for the epidemiological surveillance of bovine hypodermosis. Veterinary
Boulard, C., Quiroz, H., Kunz, M., Pruett, M. and Tarry, M. (1991). Myiases a
localisation cutanCe. DonnCes nouvelles sur la biologie, l'immunologie et
I'am6lioration de leurs contr6les. Annales de Parasitologie Humaine et
Comparee 66, 52-57.
Bourdeau, P., Klap, D.F. and Mialot, M. (1988). Myiase a Dermatobia hominis. A
propos d'un cas observe chez le chien. Recueil de Mtdicine Vkttrinaire 164,
901-906.
Bowles, V.M., Camegie, P.R. and Sandeman, R.M. (1988). Characterisation of
proteolytic and collagenolytic enzymes from the larvae of Lucilia cuprina, the
sheep blowfly. Australian Journal of Biological Research 41, 269-278.
Bowles, V.M., Grey, S.T. and Brandon, M.R. (1992). Cellular immune responses in
the skin of sheep infected with larvae of Lucilia cuprina, the sheep blowfly.
Veterinary Parasitology 44, 15 1-162.
Brain, V., Van Der Merwe, H.E. and Horak, I.G. (1983). Strobiloestrus sp. larvae in
Merino sheep. Journal of the South African Veterinary Association 54, 185-186.
Brinkmann, A. (1976). Blowfly myiasis of sheep in Norway. Norwegian Journal of
Zoology 24,325-330.
Bristow, G.A., Berland, B. and FossB, S.A. (1990). A first case of myiasis in fish.
Journal of Parasitology 76, 256257.
Broadmeadow, M., Gibson, J.E., Dimmock, C.K., Thomas, R.J. and O’Sullivan,
B.M. (1984). The pathogenesis of flystrike in sheep. Wool Technology and Sheep
Breeding 32, 28-32. -
Broce, A.B., Goodenough, J.L. and Coppedge, J.R. (1977). A wind oriented trap
for screwworm flies. Journal of Economic Entomology 70, 413-416.
Cameron, J.A., Shoukrey, N.A. and Al-Garni, A.A. (1991). Conjunctival
ophthalmomyiasis caused by the sheep nasal botfly (Oestrus ovis). American
Journal of Ophthalmology 112, 331-334.
Capelle, K.J. (1970). Studies on the life history and development of Cuterebra
polita (Diptera: Cuterebridae) in four species of rodents. Journal of Medical
Entomology 7 , 320-327.
Carpenter, T.L. and Chastain, D.O. ( 1992). Facultative myiasis by Megaselia sp.
(Diptera: Phoridae) in Texas: a case report. Journal of Medical Entomology 29,
56 1-563.
Catts, E.P. (1967). Biology of a Californian rodent bot fly Cuterebra latifrons
Coquillett (Diptera: Cuterebridae). Journal of Medical Entomology 4, 87-101.
Catts, E.P. (1982). Biology of New World bot flies: Cuterebridae. Annual Review
of Entomology 27, 318-338.
Chabaudie, N. and Boulard, C. (1992a). Effect of hypodermin A, an enzyme
secreted by Hypoderma lineatum (Insect Oestridae), on the bovine immune
system. Veterinary Immunology and Immunopathology 31, 167-177.
Chabaudie, N. and Boulard, C. (1992b). Immunomodulation of the bovine defence
system by the larval secretions of Hypoderma bovis and H . lineatum. In
“Improvements in Control Methods for Warble-fly in Cattle and Goats”
(A. Gasca, S. Hernandez, J. Martinez, and K. Pithan, eds), pp. 115-131.
Commission of the European Communities EUR 1435 1, Brussels.
Chabaudie, N. and Boulard, C. (1993). In vitro and ex vivo responses of bovine
lymphocytes to hypodermin C, an enzyme secreted by Hypoderma lineatum
(Insect Oestridae). Veterinary Immunology and Immunopathology 36, 153-162.
Chabaudie, N., Villejoubert, V. and Boulard, C. (1991). The response of cattle
vaccinated with hypodermin A to a natural infestation of Hypoderma bovis and
Hypoderma lineatum. International Journal for Parasitology 21, 859-862.
Chamberlain, W.F. and Scholl, P.J. (1991). New procedures to enhance survival of
third-instar Hypoderma lineatum (Villers) (Diptera: Oestridae) in artificial
media. Journal of Medical Entomology 28, 266-269.
Chodosh, J. and Clarridge, J. (1992). Ophthalmomyiasis: a review with special
reference to Cochliomyia hominivorax. Clinical Infectious Diseases 14,
444449.
Chopra, A., Probert, A.J. and Beer, W.E. (1985). Myiasis due to tumbu fly. Lancet
i, 1165.
Cogley, T.P. (1990). Morphology of the eggs of the rhinoceros bot flies Gyrostigma
conjugans and G . pavesii (Diptera: Gasterophilidae). International Journal of
Insect Morphology and Embryology 19, 323-326.
Cogley, T.P. (1991). Key to the eggs of the equid stomach bot flies Gasterophilus
Leach I8 17 (Diptera: Gasterophilidae) utilizing scanning electron microscopy.
Systematic Entomology 16, 125-1 33.
Cogley, T.P. and Anderson, J.R. (1983). Ultrastructure and function of the attach-
ment organ of Gasterophilus eggs (Diptera: Gasterophilidae). International
Journal of Insect Morphology and Embryology 12, 13-23.
Cogley, T.P. and Cogley, M.C. (1989). Morphology of the eggs of the human bot
fly, Dermatobia hominis (L. Jr.) (Diptera: Cuterebridae) and their adherence to
the transport carrier. International Journal of Insect Morphology and Embryol-

ogy 18, 239-248.
Colwell, D.D. and Baron, R.W. (1990). Early detection of cattle grub (Hypoderma
lineatum and H . bovis) (Diptera, Oestridae) using ELISA. Medical and
Veterinary Entomology 4, 3 5 4 2 .
Cook, D.F. and Steiner, E.C. (1990). Susceptibility of mulesing wounds to oviposi-
tion ahd strike development by the Australian sheep blowfly, Lucilia cuprina.
Austrcllian Veterinary Journal 67, 352-355.
Cooper, D.J., Pinock, D.E. and Were, S.T. (1985). Bacterial pathogens of the
Australian sheep blowfly, Lucilia cuprina. Proceedings of the 4th Australian
Conference on Grassland Ecology (ed. R.B. Chapman), pp. 236-243, Caxton
Press.
Cope, S.E. and Catts, E.P. (1991). Parahost behaviour of adult Gasterophilus
intestinalis (Diptera: Gasterophilidae) in Delaware. Journal of Medical
Entomology 28, 67-73.
Coppedge, J.R., Ahrens, E., Goodenough, J.L., Guillot, F.S. and Snow, J.W.
(1977).,Field comparisons of liver and a new chemical mixture as attractants
for the screwworm fly. Environmental Entomology 6, 66-68.
Coppedge, J.R., Broce, A.B., Tannahill, F.H., Goodenough, J.L., Snow, J.W. and
Crystal, M.M. (1978a). Development of a bait system for suppression of adult
screwworms. Journal of Economic Entomology 71, 483-486.
Coppedge, J.R., Goodenough, J.L., Broce, A.B., Snow, J.W., Crystal, M.M. and
Petersen, H.D.V. (1978b). Evaluation of the screwworm adult suppression
system (SWASS) on the island of Curacao. Journal of Economic Entomology
71,579-584.
Cork, A. (1994). Identification of electrophysiologically-active compounds for
New World screwworm, Cochliomyia hominivorax, in larval wound fluid.
Cruz, J.B., Benitez-Usher, C., Cramer, L.G., Gross, S.J. and Kohn, A.B. (1993).
Efficacy of abamectin injection against Dermatobia hominis in cattle.
Parasitology Research 79, 183-1 85.
Cunningham, E.P., Abusowa, M., Lindquist, D.A., Sadahmed, A.E. and Vargas-
T e r h , M. (1992). &e programme d'kradication de la lucilie bouchkre d'Afrique
du Nord. Revue dElevage et de Me'decine Ve'te'rinairedes Pays Tropicaux 45,
115-1 18.
Dalwitz, R. Roberts, J.A. and Kitching, R.L. (1984). Factors determining the
predominance of Lucilia cuprina larvae in blowfly strikes of sheep in southern
New South Wales. Journal of the Australian Entomological Society 23, 175-177.
Daoud, M.S., Abid, T.A. and Al-Amary, A.M. (1989). Rectal prolapse due to
Gasterophilus intestinalis larvae in a horse: a case report. Iraqi Journal of
Veterinary Sciences 2, 1-2.
Davies, L. (1990). Species composition and larval habitats of blowfly (Calliphor-
idae) populations in upland areas in England and Wales. Medical and Veterinary
Davies, W.M. (1934). The sheep blowfly problem in North Wales. Annals of
Applied Biology 21, 267-282.
Deroo, H., Jongbloet, L., Aelbrecht, M., Van Hecke, E. and Naeyaert, J.M. (1990).
Human cutaneous parasitosis: two cases of furuncular and creeping myiasis.
Dermatologica 180, 199-200.
Desbordes, O.K. and Ajogi ( 1993). Seasonal prevalence of Cephalopina titillator
myiasis in camels (Camelus dromedarius) in Sokoto State, Nigeria. Veterinary

Doby, J.M., Quilici, M., Senet, J.M., Leynia de la Jarrige, P. and Mary, C. (1987).
Evolution des anticorps dans l’hypodermose humaine. Etude en parallele par
immuno-electrophorese, ELISA et hemagglutination passive. Relations avec
hypereosinophilie sanguine et manifestations cliniques. Bulletin de la Sociktk
Franfaise de Parasitologie 5, 1 13- 122.
Dorchies, Ph., Alzieu, J.P., Bichet, H. and Chiarisoli, 0. (1989). Traitement et
prkvention de l’oestrose ovine par le Closantel. Revue de Mkdecine Vkterinaire
140, 1121-1 124.
Dorchies, Ph., Alzieu, J.P., Yilma, J.M., Donat, F., Jeanclaude, D. and Chiarisoli,
0. (1992). Prkvention de l’oestrose ovine par deux traitements au Closantel en
cours d’ktC. Revue de Mkdecine Vkterinaire 143, 45 1-455.
.Dorchies, Ph., Yilma, J.M. and Savey, J. (1993). Prevalence of lung abscesses and
interstitial pneumonia in ovine oestrosis. Veterinary Record 133, 325.
Dove, W.E. (1935). Myiasis of man. Journal of Economic Entomology 30, 29-39.
Downes, C.M., Theberge, J.B. and Smith, S.M. (1986). The influence of insects on
the distribution, microhabitat choice, and behaviour of the Burwash caribou
herd. Canadian Journal of Zoology 64, 622-629.
Drummond, R.O., Lambert, G., Smalley, H.E.Jr and Terrill, C.E. (1981). Estimated
losses of livestock to pests. In “CRC Handbook of Pest Management in
Agriculture” (D. Pimentel, ed.), pp. 11 1-127. CRC Press, Boca Raton, Florida.
Dymock, J.J. and Forgie, S.A. (1993). Habitat preferences and carcase colonisation
by sheep blowflies in the northern North Island of New Zealand. Medical and
Veterinary Entomology 7, 155-1 60.
Dymock, J.J., Peters, M.O.E., Herman, T.J.B. and Forgie, S.A. (1991). A study of
sheep blowflies at Limestone Downs sheep station in the northern Waikato, New
Zealand, over two summers. New Zealand Journal of Agricultural Research 34,
311-316.
Eagleson, J.S., Thompson, D.R., Scott, P.G., Cramer, L.G. and Barrick, R.A.
(1993). Field trials to confirm the efficacy of ivermectin jetting fluid for control
of blowfly strike in sheep. Veterinary Parasitology 51, 107-1 12.
East, I.J., Eisemann, C.H., Vuocolo, T., Pearson, R.D., Donaldson, R.A. and
Cadogan, L.C. (1992). Vaccines against blowfly strike: the effect of adjuvant
type on vaccine effectiveness. International Journal for Parasitology 22,
309-3 14.
Edwards, K.M., Meredith, T.A., Hagler, W.S. and Healy, G.R. (1984). Ophthal-
momyiasis interna causing visual loss. American Journal of Ophthalmology 97,
605-6 10.
Eggleton, P. and Belshaw, R. (1992). Insect parasitoids: an evolutionary overview.
Philosophical Transactions of the Royal Society of London B . 337, 1-20.
Egri, B. (1989). Method for evaluating the larvicidal efficacy of Strongid Plus
(Pfizer) and Eqvalan (MSD) paste against gasterophilus. Parasitologia
Hungarica 22, 109-1 14.
Eisemann, C.H., Johnson, L.A.Y., Broadmeadow, M., O’Sulivan, B.M., Donald-
son, R.A., Pearson, R.D., Vuocolo, T. and Kerr, J.D. (1990). Acquired resistance
of sheep to larvae of Lucilia cuprina, assessed in vivo and in vitro. International
Eisemann, C.H., Pearson, R.D., Donaldson, R.A., Cadogan, L.C. and Vuocolo, T.
(1993). Uptake and fate of specific antibody in feeding larvae of the sheep
blowfly, Lucilia cuprina. Medical and Veterinary Entomology 7, 177-1 85.
El-hazy, O.M.E. (1990). Wound myiasis caused by Cochliomyia hominivorax in

humans in Libya. Transactions of the Royal Sociev of Tropical Medicine and
Hygiene 84,147-748.
El-Kadery, A. and El-Begermy, M.A. (1989). Aural myiasis caused by Wohlfahrtia
magnijica. Journal of the Egyptian Society of Parasitology 19, 75 1-753.
Erzinqlioglu, Y.Z. (1987). The larvae of some blowflies of medical and veterinary
importance. Medical and Veterinary Entomology 1, 121-125.
Erzinqlioglu, Y.Z. (1989). The origin of parasitism in blowflies. British Journal of
Entomology and Natural History 2, 125-128.
Erzinqlioglu, Y.Z. and Davies, S.W. (1984). The blue-bottle Calliphora vicina R.-
D. as a parasite (primary myiasis agent), particularly on small mammals.
Naturalist 109, 3 1-34.
Escartin-Peiia, M. and Bautista-Garfias, C.R. (1993). Comparison of five tests for
the serological diagnosis of myiasis by Gasterophilus spp. larvae (Diptera:
Gasterophilidae) in horses and donkeys: a preliminary study. Medical and
Veterinary Entomology 7, 233-237.
Esser, J.R. (1991). Biology of Chrysomya megacephala (Diptera: Calliphoridae)
and reduction of losses caused to the salted-dried fish industry in south-east Asia.
Bulletin of Entomological Research 81, 3 3 4 1 .
Eyre, P., Boulard, C. and Deline, T. (1981). Local and systemic reactions in cattle
to Hypoderma lineatum larval toxin: protection by phenylbutazone. American
Journal of Veterinary Research 42, 25-28.
FA0 (1991). “The New World Screwworm Eradication Programme: North Africa
1988-1992.” Food and Agriculture Organization of the United Nations, Rome,
192 pp.
Folstad, I., Nilssen, A.C., Halvorsen, 0. and Andersen, J. (1991). Parasite avoid-
ance: the cause of post-calving migrations in Rangifer? Canadian Journal of
Zoology 69, 2423-2429.
Foster, G.G. (1989). The sheep blowfly genetic control programme in Australia.
Insect and Pest Control Newsletter FAO/IAEA, 43, 23-26.
Foster, G.G. and Smith, P.H. (1991). Genetic control of Lucilia cuprina: analysis of
field trial data using simulation techniques. Theoretical and Applied Genetics 82,
3343.
Foster, G.G., Vogt, W.G., Woodburn, T.L. and Smith, P.H. (1988). Computer
simulation of genetic control. Comparison of sterile male and field-female
killing systems. Theoretical Applied Genetics 76, 870-879.
Foster, G.G., Weller, G.L., James, W.J.., Paschalidis, K.M. and McKenzie, L.J.
(1992). Advances in sheep blowfly genetic control in Australia. In: “IAEA/FAO
International Symposium on Management of Insect Pests: Nuclear and Related
Molecular and Genetic Techniques,” Vienna.
Fox, M.T., Jacobs, D.E., Hall, M.J.R. and Bennett, M.P. (1992). Tumbu fly
(Cordylobia anthropophaga) myiasis in a quarantined dog in England.
veterinary Record 130, 100-101.
Fraiha, H., Chaves, L.C.L., Borges, I.C. and Freitas, R.B. de (1984). Miiases
humanas na Amazania - 111: Miiase pulmonar por Alouattamyia baeri (Shan-
non and Greene, 1926) (Diptera: Cuterebridae). Seperata da Revista da Funda-
ccio Sesp 29, 63-68.
French, N.P., Wall, R., Cripps, P.J. and Morgan, K.L. (1992). The prevalence,
regional distribution and control of blowfly strike in England and Wales.
French, N.P., Wall, R. and Morgan, K.L. (1994a). Blowfly strike in England and
Wales: the relationship between prevalence and farm management factors.

Medical and Veterinary Entomology 8, 5 1-56.
French, N.P., Wall, R. and Morgan, K.L. (1994b). The role of tail docking in sheep
health and productivity. Veterinary Record 134, 463-467.
French, N.P., Wall, R. and Morgan, K.L. (1994~).Ectoparasite control on sheep
farms in England and Wales; the method, type and timing of insecticide
application. Veterinary Record 135, 35-38.
Friedel, T., Hales, D.F. and Birch, D. (1988). Cyromazine-induced effects on the
larval cuticle of the sheep blowfly, Lucilia cuprina: ultrastructural evidence for a
possible mode of action. Pesticide Biochemistry and Physiology 31, 99-107.
Gabaj, M.M., Wyatt, N.P., Pont, A.C., Beesley, W.N., Awam, M.A.Q., Gusbi,
A.M. and Benhaj, K.M. (1989). The screwworm fly in Libya: a threat to the
livestock industry of the Old-world. Veterinary Record 125, 347-349.
Gall, Y., Pecquet, C., Litoux, P. and Barriere, H. (1987). Myase furonculoide a
’
Cordylobia anthropophaga examen en microscopie Clectronique a balayage.
Annals of Dermatology and Venereology 114, 59-63.
Gasca, A., Hernandez, S., Martinez, J. and Pithan, K. (eds) (1992). “Improvements
in Control Methods for Warble-fly in Cattle and Goats.” Commission of the
European Communities EUR 14351, Brussels, 144 pp.
Gassner, F.X. and James, M.T. (1948). The biology and control of the fox maggot,
Wohlfahrtia opaca (Coq.). Journal of Parasitology 34, 44-50.
Gherardi, S.G. (1977). A literature review of surgical and chemical mulesing
methods of sheep blowfly control. West Australian Department of Agriculture
Technical Bulletin No. 39.
Gherardi, S.G., Sutherland, S.S., Monzu, N. and Johnson, K.G. (1983). Field
observations on body strike in sheep affected with dermatophilosis and fleece
rot. Australian Veterinary Journal 60, 27-28.
Geigy, R. and Kauffmann, M. (1977). Experimental mechanical transmission of
Trypanosoma brucei by Auchmeromyia larvae. Protozoology 3, 103-1 07.
Gingrich, R.E. ( 1982). Acquired resistance to Hypoderma lineatum: comparative
immune response of resistant and susceptible cattle. Veterinary Parasitology 9,
233-242.
Grabbe, R.R. and Turner, J.P. (1973). Screw-worm attractants: isolation and
identification of organic compounds from bacterially inoculated and incubated
blood. Folia Entomological Mexicana 25-26, 120-1 2 1.
Graf, J-F. (1993). The role of insect growth regulators in arthropod control.
Parasitology Today 9, 471-474.
Graham, O.H. (1985). Symposium on eradication of the screwworm from the
United States and Mexico. Miscellaneous Publications of the Entomological
Society of America 62, 1-68.
Green, C.H. and Wames, M.L. (1992). Responses of female New World screw-
worm flies, Cochliomyia hominivorax, to coloured targets in the laboratory.
Medical and Veterinary Entomology 6, 103-1 09.
Green, C.H., Hall, M.J.R., Fergiani, M., Chirico, J. and Husni, M. (1993). Attract-
ing New World screwworm, Cochliomyia hominivorax, to odour-baited targets
in the field. Medical and Veterinary Entomology 7 , 59-65.
Greenberg, B. (1984). Two cases of human myiasis caused by Phaenicia sericata
(Diptera: Calliphoridae) in Chicago area hospitals. Journal of Medical
Entomology 21, 615.
Grogan, T.M., Payne, C.M., Payne, T.B., Spier, C., Cromey, D.W., Rangel, C.
and Richter, L. (1987). Cutaneous myiasis: immunohistologic and ultrastruc-
MWASlS OF HUMANS AND DOMESTIC ANIMALS 319
tural morphometric features of a human botfly lesion. American Journal of

Dermatopathology 9, 232-239.
Gruss, B. (1988). Prevention of blowfly strike on sheep by cypermethrin. Journal
of the South African Veterinary Association 59, 200-20 1.
Gruss, B. and van Rensburg, B.J. (1988). Propetamphos: a comparison of three
formulations in preventing blowfly strike in sheep. Journal of the South African
Veterihary Society 59, 77-79.
Guemni, V.H. (1988). Ammonia toxicity and alkalosis in sheep infested by Lucilia
cuprina larvae. International Journal for Parasitology 18, 79-8 1.
Guimariies, J.H. and Papavero, N. (1966). A tentative annotated bibliography of
Dermatobia hominis (Linnaeus Jr, 1781) (Diptera, Cuterebridae). Archivos de
Zoologia do Estado de Sao Paulo 14, 223-294.
G u h n d e s , J.H., Prado, A.P. and Linhares, A.X. (1978). Three newly introduced
blowfly species in Southern Brazil (Diptera: Calliphoridae). Review of Brazilian
Guimariies, J.H., Papavero, N., Pires-do-Prado, A. (1983). As miises na regiao
neotropical (identrficacao, biologia, bibliografia). Revista Brasileira de Zool&ia
1,239416.
Guilot, F.S., Coppedge, J.R., Goodenough, J.L., Ahrens, E. and Adams, T.S.
(1977). Reproductive status of female screwworms captured from hosts or in
traps. Southwestern Entomologist 2, 49-52.
Guitton, Ch. and Dorchies, Ph. (1993). etude des lames d’Oestrus ovis (Linnt
1761) en microscopie Clectronique a balayage. Revue de Mkdecine Vktkrinaire
144,687-692.
Gupta, S.C., Kumar, S. and Srivastava, A. (1982). Urethral myiasis. Tropical and
Geographical Medicine 35, 73-74.
Hadani, A., Rabinsky, R., Shimshoni, A. and Vishinsky, Y. (1971). Myiasis caused
by Wohljahrtia magnijica (Schiner) in sheep on the Golan Heights. Refuah
Veterinarith 28, 25-33.
Hadani, A., Ben Yaakov, B. and Rosen, Sh. (1989). Myiasis caused by Wohljahrtia
magnifrca (Schiner, 1862) in the Arabian camel (Camelus dromedarius) in the
Peninsula of Sinai. Revue dElevage et de Mkdecine Vktkrinaire des Pays
Tropicaux 42, 33-38.
Hadow, A.J. and Muirhead Thomson, R.C. (1937). Sheep myiasis in south-west
Scotland, with special reference to the species involved. Parasitology 29, 96-
116.
Hajek, A.E. and St Leger, R.J. (1994). Interactions between fungal pathogens and
insect hosts. Annual Review of Entomology 39, 293-322.
Hall, M.J.R. (1991). Screwworm flies as agents of wound myiasis. In “ World
Animal Review,” Special Issue “New World Screwworm: Response to an
Emergency”, October 1991, (R.D.S. Branckaert, ed.). Food and Agriculture
Organization of the United Nations, Rome.
Hall, M.J.R. and Smith, K.G.V. (1993). Diptera causing myiasis in man. In
“Medical Insects and Arachnids” (R.P. Lane and R.W. Crosskey, eds), pp.
429-469, Chapman and Hall, London.
Hall, M.J.R., Farkas, R., Kelemen, F., Hosier, M.J. and El-Khoga, J.M. (1995).
Orientation of agents of wound myiasis to hosts and artificial stimuli in Hungary.
Medical and Veterinary Entomology 9.
Hall, R.D. (1948). “The Blowflies of North America.” The Thomas Say Foundation,
Baltimore, MD.
Hall, R.D. and Townsend, L.H. (1977). The insects of Virginia No. 11. Research
Division Bulletin 123, Virginia Polytechnic Institute and State University.
Hall, R.D., Anderson, P.C. and Clark, D.P. (1986). A case of human myiasis caused
by Phormia regina (Diptera: Calliphoridae) in Missouri, USA. Journal of
Medical Entomology 23, 578-579.
Hammack, L. and Holt, G.G. (1983). Responses of gravid screwworm flies,
Cochliomyia hominivorax, to whole wounds, wound fluid and a standard blood
attractant in olfactometer tests. Journal of Chemical Ecology 9, 9 13-922.
Hammack, L., Bromel, M., Duh, F.M. and Gassner, G. (1987). Reproductive
factors affecting responses of the screwworm fly, Cochliomyia hominivorax
(Diptera: Calliphoridae), to an attractant of bacterial origin. Annals of the
Entomological Society of America 80, 775-780.
Haralampidis, S.T. ( 1987). ELISA in the seroepidemiology of parasitoses of sheep
and goats. Deltion tes Ellenikes Kteniatrikes Etaireias 38, 215-223.
' Harvey, J.T. (1986). Sheep botfly: ophthalmomyiasis externa. Canadian Journal of
Ophthalmology 21, 92-95.
Hasegawa, S., Miwata, H., Masuda, S., Naruse, H. and Ozaki, T. (1992). An
infantile case of intestinal myiasis. Acta Paediatrica Japonica 34, 87-89.
Haufe, W.O. and Nelson, W.A. (1957). Human furuncular myiasis caused by the
flesh fly Wohlfahrtia opaca (Coq.) (Sarcophagidae: Diptera). Canadian Entomol-
ogist 89, 325-327.
Hausdorfer-Scheiff, S., Bourland, A. and Pirard, Ch. ( 1993). Histopathological
aspects of myiasis. Dermatology 186, 298-300.
Hay, J. (1990). Dermatobia hominis myiasis in Leicester. Entomologist 109,
125-1 26.
Heath, A.C.G. and Bishop, D.M. (1986). Flystrike in sheep. Annual Report, 1985/86,
Wallaceville Animal Research Centre, Agricultural Research Division, 1 11-1 12.
Heath, A.C.G., Bishop, D.M. and Tenquist, J.D. (1987). The effects of artificially-
induced fly-strike on food intake and liveweight gain in sheep. New Zealand
Hendrickx, M.O., Anderson, L., Boulard, C., Smith, D.G. and Weatherley, A.J.
(1993). Efficacy of doramectin against warble fly larvae (Hypoderma bovis).
Herd, R., Strong, L. and Wardhaugh, K. (eds) (1993). Environmental impact of
avermectin usage in livestock. Veterinary Parasitology (Special Issue) 43, 344
PP.
Higgins, A.J. (1985). The camel in health and disease. 4. Common ectoparasites of
the camel and their control. British Veterinary Journal 141, 197-216.
Hjra, P.R., Hajj, B., AI-Ali, F. and Hall, M.J.R. (1993). Ophthalmomyiasis in
Kuwait: first report of infections due to larvae of Oestrus ovis before and after
the Gulf Conflict. Journal of Tropical Medicine and Hygiene 96, 241-244.
Hope, F.W. (1840). On insects and their larvae occasionally found in the human
body. Transactions of the Royal Entomological Society of London 2, 256-271.
Horak, I.G. (1987). Arthropod parasites of some wild animals in South Africa and
Namibia. Journal of the South African Veterinary Association 58, 207-21 1.
Horak, I.G. and Boomker, J. (1981). Strobiloestrus sp. larvae in cattle. Journal of
the South African Veterinary Association 52, 21 1-212.
Horak, I.G. and Snijders, A.J. (1974). The effect of Oestrus ovis infestation on
Merino lambs. Veterinary Record 94, 12-16.
Horak, I.G., Louw, J.P. and Raymond, S.M. (1971). Trials with rafoxanide. 3.
Efficacy of rafoxanide against the larvae of the sheep nasal bot fly Oestrus
ovis, LinnB, 1761. Journal of the South African Veterinary Medicine Association
42,337-339.
Humphrey, J.D., Spradbery, J.P. and Tozer, R.S. (1980). Chrysomya bezziana:
pathology of old world screw-worm fly infestations in cattle. Experimental
Hussein, M.F., Elamin, F.M., El-Taib, N.T. and Basmaeil, S.M. (1982). The
pathology of nasopharyngeal myiasis in Saudi Arabian camels (Camelus drome-
-&mius).Veterinary Parasitology 9, 253-260.
Ilchmann, G. and Splistester, H. (1982). Aerosolbedhandlung bei der oestrose der
Schafe. Tagungsbericht 197, 8 1-85.
Ilchmann, G., Betke, P., Grafe, D. and Gossing, S. (1986). Untersuchungen zur
oestrose und ihre bekiimpfung in der Mongolischen Volksrepublik. Monatshefte
f i r Veterinarmedizin 41, 128-132.
Ilchmann, G., Montag, Th., Brose, E. and Nisafi, A. (1989). Zum Proteinmuster
von Gasterophilus intestinalis - larven (Diptera: Gasterophilidae), Psoroptes
cuniculi und Chorioptes bovis (Acarida: Psoroptidae). Angewandte Parasitologie
30, 1 1 1-1 15.
Isimbekov, Zh.M. and Zhanuzakov, N.Zh. (1983). Wohlfahrtiasis of sheep in
Kazakhstan and prophylactic measures against it. In “Parazitozy sel’skokho-
zyaistvennykh zhivotnykh Kazakhstana i mery ikh preduprezhdeniya” (N.Zh.
Zhanuzakov, ed.), pp. 52-62. Kazakhskii Nauchno-Issledovatel’skii Veterinarnyi
Institut, Alma-Ata.
Jackson, H.C. (1989). Ivermectin as a systemic insecticide. Parasitology Today 5,
146-156.
Jagannath, M.S., Cozab, N. and Vijayasarathi, S.K. (1989a). Histopathological
changes in the nasal passages of sheep and goats infested with Oestrus ovis
(Diptera: Oestridae). Indian Journal of Animal Sciences 59, 87-9 1.
Jagannath, M.S., Cozab, N., Abdul Rahman, S. and Honnappa, T.G. (1989b).
Incidence of Oestrus ovis in sheep and goats. Indian Journal of Animal Sciences
59, 1216-1219.
Jagannath, M.S., Cozab, N., Abdul Rahman, S. and Honnappa, T.G. (1989~).
Serodiagnosis of Oestrus ovis infestation in sheep and goats. Indian Journal of
Animal Sciences 59, 1220-1 224.
James, M.T. (1947). The flies that cause myiasis in man. United States Department
of Agriculture Miscellaneous Publication No. 631, 175 pp.
James, M.T. (1969). A study in the origin of parasitism. Bulletin of the Entomo-
logical Society of America 15, 25 1-253.
Johnson, L.S. and Albrecht, D.J. (1993). Effects of haematophagous ectoparasites
on nestling house wrens, Troglodytes aedon: who pays the cost of parasitism.
Oikos 66,255-262.
Johnston, L.A.Y., Eisemann, C.H., Donaldson, R.A., Pearson, R.D. and Vuocolo,
T. (1992). Retarded growth of Lucilia cuprina larvae on sheep and their sera
following production of an immune response. International Journal for
Jones, C.M., Oehler, D.D., Snow, J.W. and Grabbe, R.R. (1976). A chemical
attractant for screwworm flies. Journal of Economic Entomology 69, 389-391.
Jordan, P.A., Botkin, D.B. and Wolfe, M.L. (1971). Biomass dynamics in a moose
population. Ecology 52, 147-152.
Kagei, N., Murata, I. and Kurahashi, H. (1989). An imported case of myiasis
caused by a larva of Cordylobia anthropophaga from Africa. Japanese Journal
of Tropical Medicine and Hygiene 17, 347-350.
Kalelioglu, M., Aktiirk, G., Aktiirk, F., Komsuoglu, S.S., Kuzeyli, K., Tigin, Y.,
Karaer, Z. and Bingol, R. (1989). Intracerebral myiasis from Hypoderma bovis
larva in a child. Journal of Neurosurgery 71, 929-931.
Kal’vish, T.K. ( 1990). Morphophysiological features of the entomopathogenic
fungus Tolypocladium niveum (0. Rostr.) Bissett (Deuteromycotina:
Moniliales) new to the flora of the USSR. Mikologiya i Fitopatologiya 24,
210-215.
Karter, A.J., Folstad, I. and Anderson, J.R. (1992). Abiotic factors influencing
embryonic development, egg hatching, and larval orientation in the reindeer
warble fly, Hypoderma tarandi. Medical and Veterinary Entomology 6,355-362.
Kearney, M.S., Nilssen, A.C., Lyslo, A., Syrdalen, P. and Dannevig, L. (1991).
Ophthalmomyiasis caused by the reindeer warble fly larva. Journal of Clinical
Pathology 44, 276-284.
Keech, J.P. (1981). Dermatobia hominis - in kelize. Journal of the Royal Army
Medical Corps. 127, 131-133.
Kenney, M. (1945). Experimental intestinal myiasis in man. Proceedings of the
Society for Experimental Biology and Medicine 60, 235-237.
Kettle, D.S. ( 1990). “Medical and Veterinary Entomology”. Reprinted by CAB
International, Wallingford, 658 pp.
Kim, K.C. and McPheron, B.A. (eds) (1993). “Evolution of Insect Pests: Patterns
of Variation.” John Wiley, New York, 479 pp.
Kirby, W. and Spence, W. (1815). “An Introduction to Entomology”. London,
Volume 1, edition 3, 5 19 pp.
Knipling, E.F. (1955). Possibilities of insect control or eradication through the use
of sexually sterile males. Journal of Economic Entomology 48, 459462.
Komari, K., Hara, K., Smith, K.G.V., Oda, T. and Karamine, D. (1978). A case of
lung myiasis caused by larvae of Megaselia spiracularis Schmitz (Diptera:
Phoridae). Transactions of the Royal Sociery of Tropical Medicine and Hygiene
72,467-470.
Koone, H.D. and Banegas, A.D. (1959). Biology and control of Dermatobia hominis
(L. Jr.) in Honduras. Journal of Kansas Entomological Society 32, 100-108.
Krafsur, E.S., Whitten, C.J. and Novy, J.E. (1987). Screwworm eradication in north
and central America. Parasitology Today 3, 131-137.
Kremer, M., Lenys, J., Basset, M., Rombourg, H. and Molet, B. (1970). Deux cas
de myiase a Cordylobia rodhaini contractke au Cameroun et diagnostiqute en
Alsace. Bulletin de la Sociktk de Pathologie Exotique 63, 592-596.
Kunz, S.E., Scholl, P.J., Colwell, D.D. and Weintraub, J. (1990). Use of sterile
insect releases in an IPM program for control of Hypoderma lineatum and H .
bovis (Diptera: Oestridae): a pilot test. Journal of Medical Entomology 27, 523-
529.
Lancaster, J.L. and Meisch, M.V. (1986). “Arthropods in Livestock and Poultry
Production”. Ellis Horwood, Chichester, 402 pp.
Lane, R.P., Lovell, C.R., Griffiths, W.A.D. and Sonnex, T.S. (1987). Human
cutaneous myiasis - a review and report of three cases due to Dermatobia
hominis. Clinical and Experimental Dermatology 12, 4 0 4 5 .
Laurence, B.R. and Herman, F.G. (1973). Tumbu fly (Cordylobia)infection outside
Africa. Transactions of the Royal Society of Tropical Medicine 67, 888.
Lehrer, A.Z. and Verstraeten, C. ( 1991). Expansion parasitologique et gtographi-
que de Wohlfahrtia magnijica (Schiner) (Diptera Sarcophagidae) en Roumanie.
Bulletin des Recherches Agronomiques de Gembloux 26, 563-567.
Lehrer, A.Z., Lehrer, M. and Verstraeten, C. (1988). Les myiases caustes aux
moutons de Roumanie par Wohlfahrtia magnifica (Schiner) (Diptera
Sarcophagidae). Annales de Me‘decine Ve‘te‘rinaire132, 4 7 5 4 8 1.
Leiper, J.W.G. (1951). A new approach to phenothiazine therapy in sheep. Veter-
inary Record 63, 885-889.
Lello, E. de and Boulard, C. (1990). Rabbit antibody responses to experimental
infestation with Dermatobia hominis. Medical and Veterinary Entomology 4,
303-309.
Lello, E. and Peraqoli, M.T.S. (1993). Cell-mediated and humoral immune
responses in immunized and/or Dermatobia hominis infested rabbits. Veterinary
Lello, E. de, Greg6ri0, E.A.P. and Toledo, L.A. (1985). Desenvolvimento das
Gonadas de Dermatobia hominis (Diptera: Cuterebridae). Memorias do Instituto
Oswaldo Cruz, Rio de Janeiro 80, 159-170.
Le6n. D. de and.Fox, I. (1980). Canine minima myiasis in Puerto Rico - a case
report. Journal of Agriculture of the University of Puerto Rico 64, 126-128.
Liakos, B.D. (1986). Effects of hypodermatosis on the body weight of young goats.
Bulletin of the Hellenic Veterinary Medical Society 37, 8-1 2.
Liebisch, A., Froehner, H. and Elger, D. (1983). Myiasis in sheep caused by Lucilia
sericata - an approaching problem? Tierarztliche Umschau 38, 747.
Li b o n g , P.T., Lui, H. and Buck, H.W. (1992). Cutaneous myiasis: a simple and
effective technique for extraction of Dermatobia hominis larvae. International
Journal of Dermatology 31, 657-659.
Lindquist, D.A., Abusowa, M. and Hall, M.J.R. (1992). The New World screw-
worm fly in Libya: a review of its introduction and eradication. Medical and
Veterinary Entomology 6 , 2-8.
Lonsdale, B., Tarry, D.W., Bowen, F.L. and Stansfield, D.G. (1990). Cyromazine
pour-on for the prevention of cutaneous myiasis of sheep. Veterinary Record
126,207-210.
Losson, B., Lonneux, J.F. and Pithan, K. (eds) (1993). “Improvements in the
Control Methods for Warble-fly in Cattle and Goats”. Commission of the
European Communities COST 81 1, Brussels, 127 pp.
Louw, J.P. (1989). Overberg research projects. V1. The biology and control of
Oestrus ovis in sheep in the winter rainfall areas of the Southern Cape.
Lowry, M.A. (1992). Dermatobia hominis infestation: a case report. Military
Medicine 157, 683-684.
Lukin, L.G. (1989). Human cutaneous myiasis in Brisbane: a prospective study.
Medical Journal of Australia 150, 237-240.
Lysyk, T.J.,Colwell, D.D. and Baron, R.W. (1991). A model for estimating
abundance of cattle grub (Diptera: Oestridae) from the proportion of uninfested
cattle as determined by serology. Medical and Veterinary Entomology 5,
253-25 8.
Mackerras, I.M. and Fuller, M.E. (1937). A survey of the Australian sheep
blowflies. Journal of the Council for Scientific and Industrial Research
(Australia) 10, 261-270.
Mackerras, I.M., Fuller, M.E., Austen, K.M.and Lefroy, E.H.B. (1936). Sheep
blowfly investigations: the effect of trapping on the incidence of strike in sheep.
Journal of the Council for Scientific and Industrial Research (Australia) 9,
153-162.
Mackley, J.W. and Brown, H.E. (1984). Swormlure-4: A new formulation of the
Swormlure-2 mixture as an attractant for adult screwworms Cochliomyia
hominivorax (Diptera: Calliphoridae). Journal of Economic Entomology 77,

1264-1268.
MacLeod, J. (1937). The species of Diptera concerned in cutaneous myiasis of
sheep in Britain. Proceedings of the Royal Entomological Sociery of London ( A ) ,
12, 127-133.
MacLeod, J. (1943). A survey of British sheep blowflies. Bulletin of Entomological
Research 34, 65-88.
Maeda, R., Makita, E., Segawa, M., Shibuya, T. and Ogino, M. (1990). A case of
myiasis caused by Dermatobia hominis. Japanese Journal of Tropical Medicine
and Hygiene 18, 197-201.
Maia, A.A.M. and Guimarles, M.P. (1985). Berne: susceptibilidade de bovinos,
distribuigio com outras miiases e abscessos. Arquivos Brasileiro de Medicina
Veterinaria e Zootaxonomia 37, 461-467. .
Maia, A.A.M. and Guimarges, M.P. (1986). Us0 da ivermectina no controle de
’
larvas de Dermatobia hominis (Linnaeus 178 1) (Diptera: Cuterebridae) em
bovinos de corte. Arquivos Brasileiro de Medicina Veterinaria e Zootaxonomia
38, 57-64.
Mariotti, J.M. and Vacheret, G. (1992). Les myiases conjonctivales, une pathologie
frkquente en Corse. Journal of French Ophthalmology 15, 679-682.
Mashkei, LA. (1990). Lucilia myiasis among sheep in the wooded and steppe zones
of the Ukrane. Veterinariya Kiev 65, 48-51.
McAlpine, J.F. (1989). Phylogeny and classification of the Muscomorpha. In
“Manual of Nearctic Diptera” Vol. 3. pp. 1397-1518. McAlpine J.F. (ed.),
Research Branch, Agriculture Canada, Monograph no. 32.
McGany, J.W., Gusbi, A.M., Baker, A., Hall, M.J.R. and Mughadmi, K. (1992).
Phoretic and parasitic mites infesting the New World screwworm fly, Cochlio-
myia hominivorax, following sterile insect releases in Libya. Medical and
Veterinary Entomology 6 , 255-260.
McIntyre, F.L. (1989). Myiasis. American Family Physician 39, 129-131.
McKenzie, J.A. and Anderson, N. (1990). Insecticidal control of Lucilia cuprina:
strategic timing of treatment. Australian Veterinary Journal 67, 385-386.
McMullin, P.F., Cramer, L.G., Benz, G., Jeromel, P.C. and Gross, S.J. (1989).
Control of Dermatobia hominis infestation in cattle using an ivermectin slow-
release bolus. Veterinary Record 124, 465.
Mehr, Z., Powers, N.R. and Konkol, K.A. (1991). Myiasis in a soldier returning
from Panama. Journal of Medical Entomology 28, 553-554.
Miller, D. (1939). Sheep maggot-fly problem. New Zealand survey 1937-1938.
New Zealand Journal of Science and Technology 21, 240-244.
Mpnfray, K. and Boulard, C. (1990). Preliminary evaluation of four immunological
tests for the early diagnosis of Hypoderma tarandi causing hypodennosis in
reindeer. Medical and Veterinary Entomology 4, 297-302.
Moraes, F.R., Costa, A.J., Vasconcelos, O.T. and Rocha, U.F. (1986). Ensaio
cornparativo de susceptibilidade natural de zubuinos e taurinos A larva de
Dermatobia hominis. Ars Veterinaria 2, 247-253.
Moriena, R.A., Racioppi, 0. and Lombardero, O.J. (1984). Utilizacion de la
ivermectina y decametrina como “uricidas” en bovinos de la Provincia de
Corrientes. Veterinaria Argenrina 1, 266-272.
Mourier, H. and Banegas, A.D. (1970). Observations on the oviposition and
the ecology of the eggs of Dermatobia hominis (Diptera: Cuterebridae).
Videnskabelige Meddelesler fra Dansk Naturhistorik Forening 33, 59-68.
Moya-Borja, G.E. (198 1). Effects of Macrocheles muscadomesticae (Scopoli) on
the sexual behaviour and longevity of Dermatobia hominis (L. Jr.). Revista
Brasileira de Biologia 41, 237-241.
Moya-Borja, G.E. and Borkovec, A. (1981). Sexual sterility of Dermatobia homi-
nis (Linnaeus Jr.) induced by chemosterilants. Revista Brasileira de Biologia 41,
5 1-56.
Moya-Borja, G.E., Muniz, R.A., Sanavria, A., Goncalves, L.C.B. and Rew, R.S.
(1993a). Therapeutic and persistent efficacy of doramectin against Dermatobia
Moya-Borja, G.E., Oliveira, R.A., Muniz, R.A. and Goncalves, L.C.B. (1993b).
Prophylactic and persistent efficacy of doramectin against Cochliomyia
hominivorax in cattle. Veterinary Parasitology 49, 95-1 05.
Nagakura, K., Kawauichi-Kato, Y.,Tachibana, H.,Kaneda, Y.,Shinonaga, S. and
,Kano,R. (1991). Three cases of intestinal myiasis in Japan. Journal of Infectious
’Diseases 163, 1170-1 171.
Nderagakura, F., Smail, A., Chardonnier, J. and Devauchelle, B. (1989). Les
myiases: localisations faciales. A propos d’une observation d’infestation A
Dermatobia hominis. Revue Stomatologie et Chirurgie Maxillo-Faciale 90,7-16.
Nedelchev, N.K. (1988). Distribution and causes of myiasis among farm animals.
Veterinarna Sbirka 86, 33-35. (In Bulgarian).
Noireau, F. ( 1992). Infestation by Auchmeromyia senegalensis as a consequence
of the adoption of non-nomadic life by Pygmies in the Congo Republic.
Transactions of the Royal Society of Tropical Medicine and Hygiene 86, 329.
Noms, K.R. (1990). Evidence for the multiple exotic origin of Australian
populations of the sheep blowfly Lucilia cuprina (Weidemann) (Diptera:
Calliphoridae). Australian Journal of Zoology 38, 635648.
Noms, K.R. and Murray, M.D. (1964). Notes on the screw-worm fly, Chrysomya
bezziana (Diptera: Calliphoridae), as a pest of cattle in New Guinea. Technical
Paper of the Division of Entomology, CSIRO Australia, No. 6.
Nunzi, E., Rongioletti, F. and Rebora, A. (1984). Dermatobia hominis infestation.
Postgraduate Medical Journal 60, 162-1 63.
Nunzi, E., Rongioletti, F. and Rebora, A. (1986). Removal of Dermatobia hominis
larvae. Archives of Dermatology 122, 140.
Nuorteva, P. (1987). Empty puparia of Phormia terraenovae R.D. (Diptera:
Calliphoridae) as forensic indicators. Annals Entomologica Fennica 53, 53-56.
O’Brien, D.J. and Fahey, G. (1991). Control of fly strike in sheep by means of a
pour-on formulation of cyromazine. Veterinary Record 129, 35 1-353.
Ockenhouse, C.F., Samlaska, C.P., Benson, P.M., Roberts, L.W., Eliasson, A.,
Malane, S. and Menich, M.D. (1990). Cutaneous myiasis caused by the African
tumbu fly (Cordylobia anthropophaga). Archives of Dermatology 126, 199-202.
O’Donnell, I.J., Green, P.E., Connell, J.A. and Hopkins, P.S. (1981). Immunization
of sheep with larval antigens of Lucilia cuprina. Australian Journal of Biological
Science 34,411417.
Oliveira, G.P. de (1991a). Dingmica parasitlria de bernes em bovinos 1. IncidEncia
em relaGao ao declibito. Pesquisa Agropecuhria Brasileira 26, 4 6 7 4 7 1.
Oliveira, G.P. de (1991b). Ecologia de Dermatobia hominis L. Jr. 1781 (Diptera:
Cuterebridae) na R e g l o de Sgo Carlos, Estado de SHo Paulo, Brasil, Turrialba
41, 367-375.
Oliveira, G.P. de (1991~).Dingmica parasitibia de Dermatobia hominis L. Jr.
1781, em bovinos. 11. Densidade, relaqgo entre regices corp6reas e efeito da
“vassoura” caudal. Turrialba 41, 359-366.
Oliveira, G.P. de (1991d). Epidemiologia de Dermatobia hominis (L. Jr. 1781)
(Diptera: Cuterebridae) em bovinos na regiIo de S I o Carlos, estado de S I o

Paulo. Brazilian Journal of Veterinary Research and Animal Science, S I o Paulo
28, 179-184.
Oliveira, G.P. and Alencar, M.M. (1992). Resistencia de Bovinos de seis graus de
sangue Holandes-Guzerii ao carrapato (Boophilus microplus) e ao berne (Der-
matobia hominis). Arquivos Brasileiro de Medecina Veterinaria e Zootaxonomia
42, 127-135.
Olsen, A.R., Sidebottom, T.H. and Bennett, S.G. (1993). The oriental latrine fly,
Chrysomya megacephala (Fabricius 1794) (Diptera: Calliphoridae) as an invad-
ing fly of public health importance. Bulletin of the Society for Vector Ecology 18,
133-146.
Omar, M.S. and Abdalla, R.E. (1992). Cutaneous myiasis caused by tumbu fly
larvae, Cordylobia anthropophaga in southwestern Saudi Arabia. Tropical
Medicine and Parasitology 43, 128-129.
‘O’Meara, P., Chandler, R.A., Auldist, M. and Sandeman, R.M. (1991).
Characterization of the wound exudate response during fly strike. I n “Immu-
nological Control of Blowfly Strike” (R.M. Sandeman, J.H. Arundel and E.A.
Scheurmann, eds), pp. 27-38, Australian Wool Corporation, Melbourne.
O’Meara, T.J., Nesa, M., Raadsma, H.W., Saville, D.G. and Sandeman, R.M.
(1992). Variation in skin inflammatory responses between sheep bred for resis-
tance or susceptibility to fleece rot and blowfly strike. Research in Veterinary
Science 52,205-210.
O’Rourke, F.J. (1954). Feruncular myiasis due to Wohlfahrtia vigil (Walker).
Canadian Medical Association Journal 71, 146-149.
Pampiglione, S., Schiavon, S., Candiani, G. and Fioravanti, M.L. (1991).
Osservazioni cliniche e parassitologiche su di un caso di miasi fomncolosa
disseminata da Cordylobia rodhaini nell homo in Etiopia. Parassitologia 33,
159- 167.
Pampiglione, S., Bettoli, V., Cestari, G. and Staffa, M. (1993). Furuncular myiasis
due to Cordylobia anthropophaga, endemic in the same locality for over 130
years. Annals of Tropical Medicine and Parasitology 87, 2 19-220.
Panciera, R.J., Ewing, S.A., Johnson, E.M., Johnson, B.J. and Whitenack, D.L.
(1993). Eosinophilic mediastinitis, myositis, pleuritis, and pneumonia of cattle
associated with migration of first-instar larvae of Hypoderma lineatum. Journal
of Veterinary Diagnostic Investigation 5, 226-23 1.
Pandey, V.S. (1989). Epidemiology of Oestrus ovis infection of sheep in the
highveld of Zimbabwe. Veterinary Parasitology 31, 275-280.
Pandey, V.S. and Ouhelli, H. (1984). Epidemiology of Oestrus ovis infection of
sheep in Morocco. Tropical Animal Health and Production 16, 246-252.
Pandey, V.S., Ouhelli, H. and Elkhalfane, A. (1980). Observations on the epi-
zootiology of Gasterophilus intestinalis and G . nasalis in horses in Morocco.
Pandey, V.S., Ouhelli, H. and Verhulst, A. (1992). Epidemiological observations
on Gasterophilus intestinalis and G . nasalis in donkeys from Morocco. Veter-
inary Parasitology 41, 285-292.
Papavero, N. (1977). The world Oestridae (Diptera), mammals and continental
drift. Series Entomologica 14, 240 pp., Junk, The Hague.
Pape, T. (1992). Phylogeny of the Tachinidae family-group (Diptera: Calyptratae).
Tijdschrift voor Entomologie 135, 43-86.
Patton, W.S. (1922). Notes on the myiasis-producing Diptera of man and animals.
Bulletin of Entomological Research 12, 239-261.
Payne, J.A. and Cosgrove, G.E. (1966). Tissue changes following Cuterebra
infestation in rodents. American Midland Naturalist 75, 205-2 13.
Podmogil’naya, A.P. (1983). Wohlfahrtia myiasis in sheep in the southern Urals
and its control. In “Veterinamaya Entomologiya i Akarologiya” (Nepoklonov,
A.A. ed.), pp. 177-182. Series Nauchnye Trudy Vaskhnil, Moscow.
Pokidov, 1.1. and Goncharov, A.P. (1971). Problems of the control of wohlfahrtiosis
in shekp. Veterinariya, Moscow 7 , 25-31.
Polidon“, G.A., Principato, M., Amenduni, M. and Bussani, F. (1992). Miasi
cutanea da Dermatobia hominis: aspetti zoonosici ed interesse zootecnico della
parassitosi. PRAXIS Veterinaria 13, 26-28.
Portchinsky, I.A. (1916). Wohlfahrtia magnifica, Schin., and allied Russian
species. The biology of this fly and its importance to man and domestic
animals. Memoirs of the Bureau Entomology Scientific Committee Central
Board of Lartd Administration and Agriculture Petrograd 11, 1-108 (In
Russian).
Puccini, V., Tassi, P. and Anu, E. (1983). Efficacia dell’ ivermectina iniettabile
contx’o le larvea di Oestrus ovis (L. 1761) in ovini infettati naturalmente. Atti
della Societa Italiana della Scienze Veterinarie 37, 732-734.
Puccini, V., Tassi, P., Spinto, S. and Gaggiano, G. (1987). Infection by the warble
fly Przhevalskiana silenus Brauer in Italian goats - an update. In “Warble Fly
Control in Europe. IV” (Puccini, V. and Tassi, P. eds) pp. 25-42. Symposium in
the Programme of Coordination of Research on Hypodermosis, Bari, April 9-1 1.
Qadri, S.M.H., Al-Ahdal, M.N. (1988). Cutaneous myiasis due to Dermatobia
hominis: report of a case. Annals of Saudi Medicine 8, 286-287.
Quesada, P., Navarrete, M.L. and Maeso, J. (1990). Nasal myiasis due to Oestrus
ovis larvae. European Archives of Oto-Rhino-Laryngology 247, 13 1-132.
Raadsma, H.W. (1987). Flystrike control: an overview of management and
breeding options. Wool Technology and Sheep Breeding 35, 174-185.
Raadsma, H.W. (1991). Fleece rot and body strike in Merino sheep. V. Heritability
of liability to body strike in weaner sheep under flywave conditions. Australian
Journal of Agricultural Research 42, 279-293.
Raadsma, H.W., Gilmour, A.R. and Paxton, W.J. (1989). Fleece rot and body strike
in Merino sheep. I1 Phenotypic and genetic variation in liability to fleece rot
following experimental induction. Australian Journal of Agricultural Research
40,207-220.
Rajapaksa, N. and Spradbery, J.P. (1989). Occurrence of the Old World screw-
worm fly Chrysomya bezziana on livestock vessels and commercial aircraft.
Australian Veterinary Journal 66, 94-96.
Rakusin, W. (1970). Ocular myiasis intema caused by the sheep nasal bot fly
(Oestrus ovis L.). South African Medical Journal 44, 1155-1 157.
Rapoza, P.A., Michels, R.G., Semeraro, R.J. and Green, W.R. (1986). Vitrectomy
for excision of intraocular larva (Hypoderma species). Retina 6, 99-104.
Rastegayev, Yu.M. (1984). Ecological characteristics of the warble and bot flies
attacking horses (Diptera: Oestridae, Gasterophilidae) in the desert zone of the
Caspian region. Entomologicheskoye Obozreniye 63, 455-459.
Rastegayev, Yu.M. (1988). Efficacy of ivermectin against Rhinoestrus and
Gasterophilus in horses. Izvestiya Akademii Nauk Turkmenskoi SSR, Seriya
Biologicheskikh Nauk 6, 67-69.
Ratcliffe, F.N. (1935). Observations on the sheep botfly (Lucilia sericata Meig.) in
Scotland. Annals of Applied Biology 22, 742-753.
Rawlins, S.C. (1988). Human myiasis in Jamaica. Transactions of the Royal

Society of Tropical Medicine and Hygiene 82, 77 1-772.
Rawlins, S.C. and Mansingh, A. (1987). A review of ticks and screwworms affect-
ing livestock in the Caribbean. Insect Science and Its Application 8, 259-267.
Reddy, N.R.J., Setty, D.R.L., Ranganath, L., Souza, P.E. D’ and Jagannath, M.S.
(1993). Ivermectin in cutaneous myiasis of dogs. Indian Veterinary Journal 70,
557-558.
Reichard, R.E., Vargas-TerSin, M. and Abu Sowa, M. (1992). Myiasis: the battle
continues against screwworm infestation. World Health Forum 13, 130-138.
Ribbeck, R., Ilchmann, G. and Hiepe, Th. (1984). Untersuchungen zur immun-
diagnostik der Gasterophilose. Angewandte Parasitologie 25, 78-83.
Ribeiro, P.B. and Oliveira, C.M.B. de (1983). Fase parasitiiria da Dermatobia
hominis (L. Jr., 1781) (Diptera: Cuterebridae) sobre Bovinos. Arquivos
Brasileiro de Medecina Veterinaria e Zootkonomia 35, 69 1-698.
* Ribeiro, P.B., Brum, J.G., Costa, P.R.P. and Batista, Z.R. (1989). Flutua@io
populacional de Dermatobia hominis (L. Jr., 1781) sobre bovinos no municipio
de Pelotas, RS. Arquivos Brasiliero de Medecina Veterinaria e Zootaxonomia
41, 223-231.
Ribeiro, V.L.S., Oliveira, C.M.B. de. and Alves Branco, F.de P.J. (1990). Pre-
valCncia e variaqdes mensais das larvas de Oestrus ovis (Linneus, 1761) em
ovinos no Municipio de BagC, RS, Brasil. Arquivos Brasiliero de Medecina
Veterinaria e Zootaxonornia 42, 2 1 1-22 1.
Rigndahl, 0. (1942). Contribution to the knowledge of the tachinids and muscids of
Norway. Troms) Museums Aarshefter 65, 27.
Roberts, D.S. (1967). Dermatophilosis infection. Veterinary Bulletin 17, 5 13-521.
Roche, S., Cross, S., Burgess, I., Pines, C. and Cayley, A.C.D. (1990). Cutaneous
myiasis in an elderly debilitated patient. Postgraduate Medical Journal 66,
776-777.
Rogers, C.E. and Knapp, F.W. (1973). Bionomics of the sheep bot fly, Oestrus ovis.
Environmental Entomology 2, 1 1-23.
Rognes, K. (1994). First record of the sheep green bottle fly Lucilia cuprina
(Wiedemann) from Europe (Diptera: Calliphoridae) with additional Spanish
records of Calliphoridae, Muscidae and Sarcophagidae. EOS Revista Espaiiola
de Entomologia (in press).
Roncalli, R.A. (1984a). The biology and control of Dermatobia hominis, the
tropical warble-fly of Latin America. Preventive Veterinary Medicine 2,
569-578.
Roncalli, R.A. (1984b). Efficacy of ivermectin against Oestrus ovis in sheep.
Veterinary Medicine 79, 1095-1097.
Roncalli, R.A. and Usher, C.B. (1988). Efficacy of ivermectin against Dermatobia
Roncalli, R.A., Barbosa, A. and Femandez, J.F. (1971). The efficacy of Rafoxanide
against the larval stages of Oestrus ovis in sheep. Veterinary Record 88,
289-290.
Rossi, M.A. and Zucoloto, S. (1972). Fatal cerebral myiasis caused by the tropical
warble fly, Dermatobia hominis. American Journal of Tropical Medicine and
Hygiene 22, 267-269.
Roosje, P.J., Hendrikx, W.M.L., Wisselink, M.A. and Willemse, T. (1992). A case
of a Dermatobia hominis infection in a dog in The Netherlands. Veterinary
Dermatology 3, 183-185.
Ruiz-Martinez, I., Cruz, S.M.D., Rodriguez, R., Lopez, D.M., Parra, M.S. and
Navio, F.A. (1987). Myiasis caused by Wohrfahrtia magnifica in Southern

Spain. Israel Journal of Veterinary Medicine 43, 34-41.
Ruiz-Martinez, I., Soler-Cruz, M.D., Benitez-Rodriguez, R., Perez-Jimenez, J.M.
and Diaz-Lopez, M. (1991). Myiasis caused by Wohlfahrtia magnifica in sheep
and goats in Southern Spain 11. Effect of age, body region and sex on larval
infestation. Israel Journal of Veterinary Medicine 46, 64-68.
Ryan, A.F. (1954). The sheep blowfly problem in Tasmania. Australian Veterinary
Journal 30, 109-1 13.
Sabrosky, C.W., Bennett, G.F. and Whitworth, T.L. (1989). “Bird Blow Flies
(Protocalliphora) in North America (Diptera: Calliphoridae) with Notes on the
Palearctic Species.” Smithsonian Institution Press, Washington, London, 3 12 pp.
Saleh, M.S.M. and El Sibae, M.M. (1993). Urino-genital myiasis due to Piophila
cQsei. Journal of the Egyptian Society of Parasitology 23, 737-739.
Sancho, E. (1988). Dermatobia, the neotropical warble fly. Parasitology Today 4,
242-246.
Sancho, E., Quesada, R., Madriz, L. and Boschini, C. (1989). Warble fly (Derma-
tobia hominis) in Costa Rican water buffaloes (Bubalus bubalis). BuffaloBulle-
tin 8, 58-62.
Sandeman, R.M. (1990). Prospects for the control of sheep blowfly by vaccination.
International Journal for Parasitology 20, 537-54 1.
Sandemann, R.M., Chandler, R.A., Collins, B.J. and O’Meara, T.J.(1992). Hyper-
sensitivity responses and repeated infections with Lucilia cuprina, the sheep
blowfly. International Journal for Parasitology 22, 1175-1 177.
Sauter, W. and Huber, P. (1988). Pharyngomyia picta (Meigen) (Dipt. Oestridae)
als erreger einer Ophthalmomyiasis beim Menschen. Vierteljahrsschrijt der
Natu@orschenden Gesellschaft in Zurich 133, 109-1 13.
Schaefer, C.W. (1979). Feeding habits and hosts of Calyptrate flies (Diptera:
Brachycera: Cyclorrhapha). Entomologia Generalis 5 , 193-200.
Schaller, G.B. (1972). “The Serengeti Lion: a Study of Predator-Prey Relations.”
University of Chicago Press, Chicago, London, 480 pp.
Schindler, P., Puccini, V., A m ,E. and Tassi, P. (1986). Efficacy of ivermectin and
rafoxanide against Oestrus ovis larvae in sheep. Journal of the Egyptian Society
of Parasitology 16, 1-7.
Scholl, P.J. (1990). A review of parasites, pathogens and predators of cattle grubs.
Southwestern Entomologist 15, 360-365.
Scholl, P.J. (1993). Biology and control of cattle grubs. Annual Review of
Scholl, P.J. and Barrett, C.C. (1986). Technique to extract Hypoderma sp. (Diptera:
Oestridae) larvae from the backs of cattle. Journal of Economic Entomology 79,
1125-1 126.
Scholl, P.J. and Weintraub, J. (1988). Gonotrophic development in Hypoderma
fineatumand H . bovis (Diptera: Oestridae), with notes on reproductive capacity.
Annals of the Entomological Society of America 81, 318-324.
Scholl, P.J., Colwell, D.D., Weintraub, J. and Kunz, S.E. (1986). Area-wide
systemic insecticide treatment for control of cattle grubs, Hypoderma spp.
(Diptera: Oestridae): two approaches. Journal of Economic Entomology 79,
1558-1563.
Scholl, P.J., Hironaka, R. and Weintraub, J. (1988). Impact of cattle grub (Hypo-
derma spp.) (Diptera: Oestridae) infestations on performance of beef cattle.
Journal of Economic Entomology 81, 246-250.
Scholl, P.J., Guillot, F.S. and Wang, G.T. (1992). Moxidectin: systemic activity
against common cattle grubs (Hypoderma lineatum) (Diptera: Oestridae) and

trichostrongyle nematodes in cattle. Veterinary Parasitology 41, 203-209.
Secco, V.N.D.P., Gregorio, E.A., Toledo, L.A. and Lello, E. (1992). Ultrastructure
of the ovary of Dermatobia hominis (Diptera: Cuterebridae) - 111. Gonial cell
degeneration. Memorias do Instituto Oswaldo Cruz, Rio de Janeiro 87, 315-380.
Shanahan, G.L. (1965). A review of the flystrike problem of sheep in Australia.
Journal of the Australian Institute of Agricultural Science 31, 11-24.
Sharma, L.K. (1992). Efficacy of ivermectin against nasal bots in camels (Camelus
dromedarius). Indian Veterinary Journal 69, 835-836.
Sherman, R.A. and Pechter, E.A. (1988). Maggot therapy: a review of the
therapeutic applications of fly larvae in human medicine, especially for treat-
ing osteomyelitis. Medical and Veterinary Entomology 2, 225-230.
Shewell, G.E. (1987a). Calliphoridae. In “Manual of Nearctic Diptera 2” (J.F.
McAlpine ed.), pp. 1133-1 145. Research Branch Agriculture Canada,
‘ Monograph 28.
Shewell, G.E. (1987b). Sarcophagidae. In “Manual of Nearctic Diptera 2” (J.F.
McAlpine ed.) pp. 1159-1 186. Research Branch Agriculture Canada,
Monograph 28.
Shishido, W.H. and Hardy, D.E. (1969). Myiasis of new-born calves in Hawaii.
Proceedings of the Hawaiian Entomological Society 20, 435438.
Shumilov, M.F. and Nepoklonov, A.A. (1983). Ecology and control of the reindeer
warble fly in the Magadan region, USSR. In “Veterinamaya Entomologiya i
Akarologiya” (A.A. Nepoklonov ed.), pp. 50-53. Kolos, Moscow, Russia.
Simetskii, M.A. (1980). Control of myiasis in animals. Veterinariya 5, 34-35.
Sinclair, I.J. and Wassall, D.A. (1983). Enzyme-linked immunosorbent assay for
the detection of antibodies to Hypoderma bovis in cattle. Research in Veterinary
Science 34, 25 1-252.
Sinclair, I.J., Tarry, D.W. and Wassall, D.A. (1990). Reduction in UK warble
infestation levels: serological survey, 1990. Veterinary Record 127, 285.
Singh, T.S. and Rana, D. (1989). Urogenital myiasis caused by Megaselia scalaris
(Diptera: Phoridae): a case report. Journal of Medical Entomology 26, 228-229.
Singh, I., Gathwala, G., Yadav, S.P.S., Wig, U. and Jakhar, K.K. (1993). Myiasis
in children: the Indian perspective. International Journal of Pediatric Otorhi-
nolaryngology 25, 127-13 1.
Smith, K.G.V. (1986). “A Manual of Forensic Entomology”. British Museum
(Natural History), London and Cornell University Press, Ithaca, 205 pp.
Smith, D.R. and Clevenger, R.R. (1986). Nosocomial nasal myiasis. Archives of
Pathology and Laboratory Medicine 110, 439440.
Spqadbery, J.P. (1981). A new trap design for screw-worm fly studies. Journal of
the Australian Entomological Society 20, 151-153.
Spradbery, J.P. (1991). “A Manual for the Diagnosis of Screw-worm Fly.” CSIRO
Division of Entomology, Canberra, 64 pp.
Spradbery, J.P. (1994). Screw-worm fly: a tale of two species. Agricultural Zoology
Reviews 6, (in press).
Spradbery, J.P. and Sands, D.A. (1981). Larval fat body and its relationship to
protein storage and ovarian development in adults of the screw-worm fly
Chrysomya bezziana. Entomologia Experimentalis et Applicata 30, 116-1 22.
Spradbery, J.P. and Owen, I.L. (1990). Efficacy of closantel against infestations of
screw-worm Chrysomya bezziana. Australian Veterinary Journal 67, 340.
Spradbery, J.P. and Kirk, J. (1992). Incidence of Old World screw-worm fly in the
United Arab Emirates. Veterinary Record 127, 33.
MWASIS OF HUMANS AND DOMESTIC ANIMALS 33 1
Spradbery, J.P. and Vogt, W.G. (1993). Mean life expectancy of Old World
screwworm fly, Chrysomya bezziana, inferred from the reproductive age-struc-
ture of native females caught on swormlure-baited sticky traps. Medical and
Veterinary Entomology 7 , 147-1 54.
Spradbery, J.P., Tozer, R.S., Drewett, N. and Lindsey, M.J. (1985). The efficacy
of ivermectin against larvae of the screw-worm fly (Chrysomya bezziana)
Australian Veterinary Journal 62,3 11-3 14.
Spradbery, J.P., Tozer, R.S., Robb, J.M. and Cassells, P. (1989). The screw-worm
fly Chrysomya bezziana Villeneuve (Diptera: Calliphoridae) in a sterile release
trial in Papua New Guinea. Researches on Population Ecology 31, 353-366.
Spradbery, J.P., Tozer, R.S. and Pound, A.A. (1991). The efficacy of insecticides
against larvae of the screw-worm fly (Chrysomya bezziana). Australian
Veterinary Journal 68,338-342.
Spratt, D.M. (1984). The occurrence of Cephalopina titillator (Clark) (Diptera:
Oestridae) in camels in Australia. Journal of the Australian Entomological
Society 23, 229-230.
Steelman, C.D. (1976). Effects of external and internal arthropod parasites on
domestic livestock production. Annual Review of Entomology 21, 155-178.
Stevens, J.D., McCartney, A.C.E. and Howes, R. (1991). Oestrus ovis ophthalmo-
myiasis acquired in the UK: case report and scanning electron microscopic
study. British Journal of Ophthalmology 75, 702-703.
Strong, L. (1992). Avermectins: a review of their impact on insects of cattle dung.
Bulletin of Entomological Research 82,265-274.
Strong, L. and Wall, R. (1990). The chemical control of livestock parasites:
problems and alternatives. Parasitology Today 6,29 1-296.
Sutherst, R.W., Spradbery, J.P. and Maywald, G.F. (1989). The potential geo-
graphical distribution of the Old World screw-worm fly, Chrysomya bezziana.
Syrdalen, P., Nitter, T. and Mehl, R. (1982). Ophthalmomyiasis interna posterior:
report of case caused by the reindeer warble fly larva and review of previous
reported cases. British Journal of Ophthalmology 66,589-593.
Tantawi, T.I. and Greenberg, B. (1993). Chrysomya albiceps and C. rufrfacies
(Diptera: Calliphoridae):contribution to an ongoing taxonomic problem. Journal
of Medical Entomology 30,646-648.
Tarry, D.W. (1986). progress in warble fly eradication. Parasitology Today 2,
111-1 16.
Tarry, D.W. (1994). Hypodermosis in Britain: a fragile success. Tenth Meeting of
the European Hypodermosis Scientijc Working Group (in press).
Tarry, D.W., Sinclair, I.J. and Wassall, D.A. (1992). Progress in the British
hypodermosis eradication programme: the role of serological surveillance.
Veterinary Record 131,3 10-3 12.
Tassi, P., Puccini, V. and Giangaspero, A. (1987). Efficacy of ivermectin against
goat warbles (Przhevalskiana silenus Brauer). Veterinary Record 120,421.
Tassi, P., Puccini, V. and Giangaspero, A. (1989). Infection by the warble fly
Przhevalskiana silenus Brauer, 1858, in the Italian goats: an update up to 1988.
Parassitologia 31, 239-250.
Taylor, D.B. (1992). Rearing systems for screwworm mass production. In
Advances in Insect Rearing for Research and Pest Management (T.E.
Anderson and N.C. Leppla, eds), pp. 393403. Westview Press, Boulder, CO.
Tenquist, J.D. (1977). Some ectoparasites of veterinary importance. New Zealand
Journal of Entomology 6, 285-289.
Tenquist, J.D. and Wright, D.F. (1976). The distribution, prevalence and economic
importance of blowfly strike in sheep. New Zealand Journal of Experimental
Agriculture 4, 291-295.
Thomas, D.B. Jr. (1987). Incidence of screwworm (Diptera: Calliphoridae) and
Torsalo (Diptera: Cuterebridae) myiasis on the Yucatan Peninsula of Mexico.
Journal of Medical Entomology 24: 498-502.
Thomas, D.B. (1988). The pattern of Dermatobia (Diptera: Cuterebridae) myiasis
in cattle in Tropical Mexico. Joltrnal of Medical Entomology 25, 131-135.
Thomas, D.B. (1993a). Behavioural aspects of screwworm ecology. Journal of the
Kansas Entomological Society 66, 13-30.
Thomas, D.B. (1993b). Fecundity and oviposition in laboratory colonies of the
screwworm fly (Diptera: Calliphoridae). Journal of Economic Entomology 86,
1464-1472.
’
Thomas, D.B. and Mangan, R.L. (1989). Oviposition and wound visiting behaviour
of the screwworm fly, Cochliomyia hominivorax (Diptera: Calliphoridae).
Thomas, D.B. and Pruett, J.H. (1989). Antibodies to antigens of Cochliomyia
hominivorax (Coquerel) (Diptera: Calliphoridae) in the sera of infested sheep.
A serological technique for the detection of screwworm myiasis. Journal of
Entomological Science 25, 143-149.
Thomas, D.B. and Pruett, J.H. (1992). Kinetic development and decline of anti-
screwworm (Diptera: Calliphoridae) antibodies in serum of infested sheep.
Journal of Medical Entomology 29, 870-873.
Tillyard, R.J. and Seddon, H.R. (1933). The sheep blowfly problem in Australia.
Report No. 1 of the Joint Blowfly Committee. Council for Scientific and
Industrial Research, Australia Pamphlet, No. 37.
Torr, S.J. and Hall, M.J.R. (1992). Odour-baited targets to control New World
screwworm (Cochliomyia hominivorax (Coquerel): Diptera: Calliphoridae): a
preliminary study. Bulletin of Entomological Research 82, 4 1 7 4 2 3 .
Townend, C. (1987). Sheep strike and mulesing. Parasitology Today 3, 68-69.
Townsend, C.H.T. (1935). “Manual of Myiology. Part 11: Muscoid Classification
and Habits.” Charles Townsend and Filhos, Silo Paulo, Brazil, 296 pp.
Uttamchandani, R.B., Trigo, L.M., Poppiti, R.J., Rozen, S. and Ratzan, K.R.
( 1989). Eosinophilic pleural effusion in cutaneous myiasis. Southern Medical
Journal 82, 1288-1 29 1.
Uribe, L.F., McMullin, P.F., Cramer, L.G. and Amaral, N.K. (1989). Topically
applied ivermectin: efficacy against Torsalo (Diptera: Cuterebridae). Journal of
‘Vinogradova, E.B. (1986). Geographical variation and ecological control of
diapause in flies. In “The Evolution of the Insect Life Cycle” (F. Taylor and
K. Karban, eds), pp. 3 5 4 7 , Springer, New York.
Vogt, W.G. and Havenstein, D.E. (1974). A standardized bait trap for blowfly
studies. Journal of the Australian Entomological Society 13, 249-253.
Vogt, W.G. and Morton, R. (1991). Estimation of population size and survival of
sheep blowfly, Lucilia cuprina, in the field from serial recoveries of marked flies
affected by weather dispersal and age-dependent trappability. Researches on
Population Ecology 33, 141-163.
Vogt, W.G., Woodburn, T.L., Morton, R. and Ellem, B.A. (1983). The analysis
and standardisation of trap catches of Lucilia cuprina (Weidemann) (Diptera:
Calliphoridae). Bulletin of Entomological Research 73, 609-617.
Vogt, W.G., Woodburn, T.L. and van Gerwen, A.C.M. (1985a). The influence of
&yte resorption on ovarian development rates in the Australian sheep blowfly,

Lucilia cuprina. Entomologial Experimentalis et Applicata 39, 85-99.
Vogt, W.G., Woodburn, T.L., Morton, R. and Ellem, B.A. (1985b). The influence
of weather and time of day on trap catches of males and females of Lucilia
cuprina (Weidemann) (Diptera: Calliphoridae). Bulletin of Entomological
Research 75, 315-319.
Wall, R. (1993). The reproductive output of the blowfly Lucilia sericata. Journal of
Insect Physiology 39, 743-750.
Wall, R., French, N. and Morgan, K. (1992a). Blowfly species composition in
sheep myiasis in Britain. Medical and Veterinary Entomology 6, 177-178.
Wall, R., Green, C.H., French, N. and Morgan, K.L. (1992b). Development of an
attractive target for the sheep blowfly Lucilia sericata. Medical and Veterinary
Entomology 6 , 67-74.
Wall, R., French, N. and Morgan, K.L. (1992~).Effects of temperature on the
development and abundance of the sheep blowfly Lucilia sericata (Diptera:
Calliphoridae). Bulletin of Entomological Research 82: 125-1 3 1.
Wall, R., French, N. and Morgan, K.L. (1993a). Predicting the abundance of the
sheep blowfly Lucilia sericata. Bulletin of Entomological Research 83,431 4 3 6 .
Wall, R., French, N. and Morgan, K.L. (1993b). Sheep blowfly population control:
development of a simulation model and analysis of management strategies.
Journal of Applied Ecology 30,143-75 1.
Wardhaugh, K.G. and Dallwitz, R. (1984). Covert fly strike. Wool, Technology and
Sheep Breeding 32, 15-19.
Wardhaugh, K.G. and Morton, R. (1990). The incidence of flystrike in sheep
in relation to weather conditions, sheep husbandry and the abundance of the
Australian sheep blowfly Lucilia cuprina (Weidemann) (Diptera: Calliphoridae).
Australian Journal of Agricultural Research 41, 1155-1 167.
Wardhaugh, K.G., Read, P. and Neave, M. (1984). A sticky-trap for studying the
spatial distribution of the Australian sheep blowfly Lucilia cuprina. Australian
Veterinary Journal 60, 132.
Wardhaugh, K.G., Vogt, W.G., Dallwitz, R. and Woodburn, T.L. (1989). The
incidence of flystrike in relation to sheep susceptibility and the abundance of
the blowfly Lucilia cuprina (Weidemann) (Diptera: Calliphoridae). General and
Applied Entomology 21, 11-16.
Washburn, R.H., Klebesadel, L.J., Palmer, J.S., Luick, J.R. and Bleicher, D.P.
(1980). The warble-fly problem in Alaska reindeer. Agroborealis 12, 23-28.
Waterhouse. D.F. and Paramanov. S.J. (1950). The status of the two sDecies of
Lucilia attacking sheep in Australia. Australian Journal of Scientific Re'search 3,
310-337.
Watts, J.E., Muller, M.J., Dyce, A.L. and Noms, K.R. (1976). The species of flies
reared from struck sheep in south-eastern Australia. Australian Veterinary
Journal 52, 488489.
Watts, J.E., Dash, K.M. and Lisle, K.A. (1978). The effect of antihelmintic
treatment and other management factors on the incidence of breech strike in
Merino sheep. Australian Veterinary Journal 54, 352-355.
Watts, J.E., Murray, M.D. and Graham, N.P.H. (1979). The blowfly strike problem
of sheep in NSW. Australian Veterinary Journal 55, 325-334.
Wegner, Z., Kudrewicz, L. and Zwierz, C. (1986). A second case of tropical
cutaneous myiasis in a sailor from the Polish merchant marine. Bulletin of the
institute of Maritime and Tropical Medicine in Gdynia 37, 5 7 4 1 .
Wegner, Z., Ozga, B. and Piatkowska, W. (1992). Nowy przypadek muszycy
skdmej z terenu Ameryki Poludniowej spowodowany przez larwe Dermatobia

hominis (L. Jr.). Polski Tygodnik Lekarski 47, 810-81 1.
Weisbroth, S.H., Wang, R, and Scher, S. (1973). Cuterebra buccata: immune
response in myiasis of domestic rabbits. Experimental Parasitology 34, 22-3 1.
Wells, J.D. (1991). Chrysomya megacephala (Diptera: Calliphoridae) has reached
the continental United States: review of its biology, pest status and spread
around the world. Journal of Medical Entomology 28, 471-473.
Wells, J.D. and Greenberg, B. (1992). Interaction between Chrysomya rufifacies
and Cochliomyia macellaria (Diptera: Calliphoridae): the possible consequences
of an invasion. Bulletin of Entomological Research 82, 133-137.
Whitten, M.J. (1979). The use of genetically-selected strains for pest replacement
or supression. In “Genetics in Relation to Insect Management” (M.A. Hoy and
J.J. McKelvey eds), pp. 3 1 4 0 . Rockerfeller Foundation, New York.
Whitten, M.J., Foster, G.G., Vogt, W.G., Kitching, R.L., Woodbum, T.L. and
L
Konovalov, C. (1977). Current status of genetic control of the Australian
sheep blowfly, Lucilia cuprina Weidernann (Diptera: Calliphoridae). Proceed-
ings of the XV International Congress of Entomology, Washington DC 1976,
pp. 129-139.
Whitworth, T.L. and Bennett, G.F. (1992). Pathogenicity of larval Protocalliphora
(Diptera: Calliphoridae) parasitizing nestling birds. Canadian Journal of
Zoology 70, 2 1 8 4 219 1.
Willadsen, P., Eisemann, C.H. and Tellam, R.L. (1993). “Concealed” antigens:
expanding the range of immunological targets. Parasitology Today 9, 132-1 35.
Williams, H. (1977). Modifications of the west Australian blowfly (Diptera:
Calliphoridae) trap for population studies. Journal of Economic Entomology
77, 806-809.
Wilson, J.A. and Heath, A.C.G. (1994). Resistance to two organophosphorus
insecticides in New Zealand populations of the Australian sheep blowfly,
Lucilia cuprina. Medical and Veterinary Entomology 8, 23 1-237.
Wood, D.M. (1987). Oestridae. In “Manual of Nearctic Diptera 2” (J.F. McAlpine,
ed.), pp. 1147-1 158. Research Branch Agriculture Canada, Monograph 28.
Yilma, J.M. and Dorchies, Ph. (1991). Epidemiology of Oestrus ovis in southwest
France. Veterinary Parasitology 40, 3 15-323.
Yilma, J.M. and Dorchies, Ph. (1993). Essais d’infestations experimentales de
l’agneau par des larves 1 d’Oestrus ovis. Bulletin de la Socie‘tk Francaise de
Parasitologie 11, 43-47,
Zakson, M., Hink, W.F. and MacKichan, J.J. (1992). Fate of the benzoylphenyl
urea CGA-184699 in the cat flea Ctenocephalides felis. Pesticide Science 35,
117-125.
Zayed, A.A. and Hilali, M. (1993). Studies on Rhinoestrus purpureus larvae
‘infesting donkeys in Egypt: localization and migratory route. Journal of Equine
Zayed, A.A., Hilali, M. and El Metenawy, T.M. (1993). Studies on Rhinoestrus
purpureus (Diptera: Oestridae) larvae infesting donkeys (Equus asinus) in
Egypt: incidence and seasonal variations. Journal of Equine Veterinary Science
13, 4 6 4 9 .
Zeleddn, R. and Silva, S. (1987). Attempts to culture the parasitic stage of
Dermatobia hominis (L. Jr.) in vitro (Diptera: Cuterebridae). Journal of Para-
sitology 73, 907-909.
Zeltser, R. and Lustmann, J. (1988). Oral myiasis. International Journal of Oral
and Maxillofacial Surgery 17, 288-289.
Zumpt, F. (1965). “Myiasis in Man and Animals in the Old World.” London,
Butterworths, 267 pp.
Parasitism and Parasitoidism in Tarsonemina
(Acari: Heterostigmata) and Evolutionary
Considerations*
Marek Kaliszewski
Brigharn Young University, Provo, Utah, USA
FranGoise Athias-Binche
Laboratoire Arago, Universitk Paris VIICNRS UA I 17,

66650 Ban.yuls sur Mer, France
and
Evert E. Lindquist
Centre for Land and Biological Resources Research,

Agriculture Canada, K.W. Neatby Building, CEF,
Ottawa, Ontario KIA OC6, Canada
1. Introduction .
....... .... ....... ...
.. ....... 336
2. Phoresy, a Pre-adaptation for Parasitism and Parasitoidism? ....... 337
3. Physogastry, a Pre-adaptation for Parasitoidism? . . . ......... 341
* This paper was initiated by the first and second authors. This collaboration was suddenly
interrupted by the tragic death of Dr Marek Kaliszewski in an automobile accident in
October 1992. Thereafter, the third author kindly agreed to augment and complete the
manuscript. The world of acarology lost in M. Kaliszewski one of its most dynamic and
innovative young scientists. This paper is dedicated to his memory.
ADVANCES IN PARASITOLOGY VOL 35 Copyright Q 1995 Academic Press Limited

ISBN 0-12-0317354 AN rights of reproduction in any form reserved
336 MAREK KALISZEWSKI ET AL
4. Distribution of Physogastry, Parasitism and Parasitoidism among

Tarsonemina .............................. 341
4.1. Superfamily Dolichocyboidea . . . . . . . . . . . . . . . . . . . . 343
4.2. Superfamily Trochometridioidea ................ .. 345
4.3. Superfamily Scutacaroidea . . . . . . . . . . . . . . . . . . . . . 346
4.4. Superfamily Pygmephoroidea . . . . . . . . . . . . . . . . . . . . 348
4.5. Superfamily Pyemotoidea . . . . . . . . . . . . . . . . . . . . . . 349
4.6. Superfamily Tarsonemoidea .................... 354
5. Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 358
Acknowledgement . . . . . . . . . . . . . . . . . . . . . . . . . . . . 360
References ............................... 360
1. INTRODUCTION
The Tarsonemina, within the subcohort Heterostigmata and suborder

Prostigmata of the Acari, constitute a relatively highly derived group of
small mites (c. 140450 pm) among Actinotrichida (Lindquist, 1986).
Their development is abbreviated, the adult emerging directly from the
larva or egg without having passed through active nymphal instars,
whereas other Actinotrichida develop through a larva and usually two or
three nymphal instars. The chelicerae of actively feeding forms are stylet-
like, adapted for piercing and sucking fluid contents from fungi, plants, or
invertebrates. Nearly all Tarsonemina are arrhenotokous (fertilized eggs
produce diploid females and unfertilized eggs produce haploid males), a
few are thelytokous, but no diplo-diploid or pseudoarrhenotokous species
exists (Kaliszewski and Wrensch, 1993). Males are so attuned and special-
ized for such precopulatory behavior that, in many families, they do not
feed.
All superfamilies of the group contain species which are associated with
arthropods and, in at least one genus of Pygmephoridae, with vertebrates.
These forms probably derived from free-living fungivorous ancestors,
though the Tarsonemina lineage as a whole ultimately arose from a
free-living predatory group, the Tarsocheyloidea, at the base of the
Heterostigmata (Lindquist, 1976). Physogastry, the extensive enlargement
of a female’s body during feeding before reproduction, is known to occur
among some or all known taxa of each of the six superfamilies of
Tarsonemina recognized here. Also, adaptations to associate with other
animals apparently occurred independently one or more times within each
of these six superfamilies. In many cases, these mites originally utilized
heterospecific carriers for phoretic dispersal, but remained free-living in
the nest or habitat of their host. However, in the course of evolution
towards host/phoront specificity, some groups gradually adapted to a
PARASITISM AND PARASlTOlDlSM IN TARSONEMINA 337
more closely symbiotic, sometimes parasitic or parasitoid, way of life

with their hosts.
This revue presents some possible routes of evolution from phoresy and
physogastry towards parasitism and parasitoidism, and the distribution of
parasites and parasitoids, among the superfamilies and families of Tarso-
nemina. For clarity, the concept of parasitism, as used here, is restricted to
use of animals as hosts, and the distinction between animal parasitism and
parasitoidism is discussed below. We agree with Price (1977) that plant-
feeding mites are also parasites of their host. However, his argument could
then be extended to fungivorous and algivorous mites as parasites of their
host, which would cloud the thrust of our account.
2. PHORESY, A PRE-ADAPTATION FOR PARASITISM AND

PAW~SITOIDISM?
Phoresy is a phenomenon in which one animal (the phoretic or phoront)

actively seeks out and attaches to an animal of another species for dis-
persal, during which time the phoront becomes quiescent, ceasing feeding
and ontogenesis (Farish and Axtell, 1971). Phoront activity commences
again beginning with detachment, induced by stimuli originating from
either its carrier or the microhabitat (Athias-Binche, 1990).
The adaptive traits of phoresy may be segregated as follows (Athias-
Binche, 1991):
initiation of phoretic behavior;
active host seeking;
recognition of attractant signals;
specificity of attachment to host surface;
quiescence;
recognition of signals to disembark and, if needed,
synchronization with the host’s life cycle.
Some of these behaviors are analogous to those of parasites, especially
those concerning host-seeking, recognition of its signals, and specificity of
the sites of attachment. Some are also similar to those of parasitoids,
namely, host-seeking, recognition of its signals, and synchronization.
Mere phoresy itself can be regarded as a peculiar case of parasitism
because the presence of the traveller may disturb the host’s activity.
This is apparent in large mites, which often regulate their numbers as
phoronts to prevent harmful effects to the carrier, especially its flying
ability, which could endanger the success of the mites’ migration (Binns,
1974; Athias-Binche, 1990).
Phoretic behavior may be facultative or obligatory (Athias-Binche,

1990). Facultative phoresy, i.e., phoresy induced by density-dependent
factors or changes in environmental conditions, is frequently encountered
in unpredictable or transient habitats. For instance, phoresy is facultative in
Pediculaster flechtmanni (Wicht), a fungivorous pygmephorid which
invades mushroom profusions (Cross and Kaliszewski, 1988). The authors
documented a pronounced dimorphism between phoretic and non-phore-
tic females, which differ from one another in behavior and structure. The
proportion of phoretic females, termed “phoretomorphs” (Moser and
Cross, 1975), increased as the culture aged, probably related to habitat
deterioration. The appearance of the phoretic phenotype is environmen-
. tally, conditionally, cued (Cross er al., 1981; Athias-Binche, 1994).
Because of the activities of the non-phoretic morph, which involve all
sorts of movement, we avoid a common reference to it as the “seden-
tary” morph; some authors call it the “normal” morph, which perhaps
gives an undesirable connotation to the phoretic morph as “abnormal”.
Among astigmatic mites, the analogous “normal” morph is called the
“homeomorph” (Houck and O’Connor, 199 1).
Among the taxa of parasites and parasitoids in the superfamily
Tarsonemoidea, only one adult female morph is known. However, among
the all-parasitoid superfamily Pyemotoidea, only one adult female morph is
known in the Acarophenacidae, but female dimorphism is recorded among
at least some members of the Pyemotidae. In this superfamily, it is
problematic whether ancestrally there were two female morphs, with
the non-phoretic morph being suppressed secondarily in the Acarophena-
cidae, or whether ancestrally there was one female morph, with female
dimorphism arising secondarily within the Pyemotidae.
In the order Anactinotrichida, Uropodina also exhibit non-phoretic and
phoretic morphs, but in the deutonymphal stage. In ephemeral habitats, the
whole generation of occupants has to migrate, and only phoretic morphs
are observed. On the other hand, in more permanent habitats, phoretic and
non-phoretic morphs coexist, the latter ensuring the reproduction and
dynamics of the local deme (Athias-Binche et al., 1993). The case may
be similar in Tarsonemina, as dimorphism seems to be present mostly in
free-living, facultatively phoretic species, and more rare in forms taking
their resources from highly transient habitats.
Obligatory phoresy is seasonal or cyclical. Seasonal phoresy, which
takes place usually once a year during the appropriate season in free-
living forms (Fashing, 1975; Athias-Binche, 1984), seems not to have
been recorded in Tarsonemina. Cyclical phoresy occurs when mite dev-
elopment is correlated with the life cycle of the host. True cyclical phoretic
populations inhabit specifically the nest of their host, and the association is
often highly specific, steno- or monoxenous. In the course of evolution
from a simple dispersal behavior to more specific associations, the phoront

progressively depends either on the host (parasitism, parasitoidism, specific
micropredation), or on its nest resources (commensalism, mutualism)
(Athias-Binche, 1990). Many Tarsonemina are cyclical, stenoxenous,
non-parasitic associates. Mutualism may result in a cleaning symbiosis
(Dindal, 1975), the mite feeding on debris or potential pathogens such as
fungi. This is the case with Scutacarus acarorum (Goeze), associated with
holarctic bumblebees, which specifically feeds on the mold of Histio-
plasma sp. in bees’ nests (Schousboe, 1986). The situation may be similar
with the pygmephorid, Petalomium fimbrisetum Ebermann & Rack, which
inh@its the nests of the ant, LasiusJlavus (Fabricius), in Europe (Ebermann
and Rack, 1982). The pygmephorid, Parapygmephorus costaricanus Rack
& Eickwort, associated with the bee, Agapostemon nasutus Smith, in
Costa Rica, feeds on the fecal substrate produced by its host larvae,
together with its eventual fungal contaminants (Rack and Eickwort,
1980). Many phoretic species of Tarsonemus and Heterotarsonemus are
apparently fungivorous under bark in the galleries of their beetle
associates, with no parasitic relationship with their carriers (Lindquist,
1969a, 1970a, 1975).
For parasitoids, synchrony with the host phenology is vital to survival of
the population because the host activity or progeny provide resources for
the mites, and also the young emerging winged adults are indispensable
agents for the mites’ disperal. Thus, the mite life cycle must be controlled
by some events affecting host development. To synchronize with the longer
generation times of their hosts, adult female mites undergo an apparently
non-feeding, prephoretic period which continues during the inactive phore-
tic period; such behavior patterns are presumed to be genetically fixed
(Lindquist, 1969b, c). In the tarsonemid genus fponemus, for example
(described further below), the parasitoid female produces a new genera-
tion of offspring that develop to adulthood without feeding. The new adult
females of this generation endure an extended, non-feeding period, waiting
for emergence of young adult bark beetles of the next generation of hosts,
and this period continues while they are quiescent phoronts on these hosts
(Lindquist, 1969~).The host activities, especially excavation of galleries,
copulation, oviposition, and imaginal molting, act as cues for such mite
behaviors as detachment, parasitoid questing and feeding, reproduction,
and initiation of phoretic attachment.
Phoresy appears to have favored evolution towards parasitism of animals
(but not of plants) in some groups of mites. An obligate, stenoxenous,
phoretic species, whose development is synchronized with that of its
phoronthost, may gradually evolve towards parasitism through paraphagy
(Athias-Binche and Morand, 1993). Phoresy, which may probably be
euryxenous at its origin, seems to have evolved towards more specific
relationships. Even in the most euryxenous forms, the individuals that

cannot recognize an adequate host will be eliminated, along with their
genotypes. These strong pressures rapidly select the most competitive
lineages and, in doing so, lead to isolating mechanisms by host/phoront
specificity. Finally, some species cannot survive in the absence of specific
hosts, and this process may lead, orthoselectively, to a “dead end”, such as
with steno- or monoxenous parasites. Then, phoresy may disappear in the
course of evolution towards permanent symbiosis (Athias-Binche, 1990).
For mites of the suborder Acaridida, or Astigmata, Houck and O’Connor
(1991) have documented that the phoretic deutonymph, or hypopus, is to
some extent parasitic in some species.: Although the deutonymph of
- Hemisarcoptes cooremani (Thomas) is typical of hypopi in having a
trophically functionless gnathosoma, it can nevertheless acquire liquid
nutriment, through the action of the suctorial disks on its posteroventral
attachment plate, from its beetle host, Chilocorus cacti (L.). The liquid is
haemolymph, reflexed by the host as an anti-predatory deterrent, which
frequently bathes the subelytral area where the mites are attached, but does
not harm them. As attachment to the host may continue even when fresh
external resources are plentiful, and as the hypopi visibly swell before
disembarking from the host, Houck and O’Connor (1991) regarded this
as strong evidence for a phoretic association evolving to a parasitic one.
In other lineages, phoresy presumably favored evolution toward a para-
sitoid way of life, through either predaceous or fungivorous steps. Para-
sitoidism is frequent in Tarsonemina, yet it has not been recorded among
other groups of mites. In some parasitoid Tarsonemina, development is
further abbreviated, the larva being regressed in the egg. In general, the
adult female mite is phoretic on an adult host, and her disembarkment is
induced by the host’s mating or reproductive activities. As noted by
Lindquist (1983), parasitoid mites are ectoparasitoids, and only the adult
female instar attacks and feeds on the host; in contrast, with insect para-
sitoids, the adult female locates its hosts, but its larval progeny feed on
them. True parasites usually do not kill their host, whereas a parasitoid’s
attack, usually on an egg or on one immature instar of the host, leads
relatively quickly to host death. Predation differs from parasitoidism in that
the biomass of the predator(s) approaches or exceeds that of the prey,
whereas parasitoids are usually much smaller than their hosts. Also, pred-
ators are usually less specific in prey choice, whereas parasitoids are often
stenoxenous, or even monoxenous. However, more important distinctions
than relative size or specificity are that a predator needs several to many
prey for it (or one of its progeny) to complete development to adulthood,
and it usually feeds during more than one stage of its development, whereas
a parasitoid needs but one host for one or more of its progeny to complete
PARASITISM AND PARASlTOlDlSM IN TARSONEMINA 34 1
development, and it feeds during only one stage of its development (i.e., as
a larva but not as an adult, or vice versa) (Lindquist, 1983).
3. PHYSOGASTRY, A PRE-ADAPTATION FOR PARASITOIDISM?
Evolution towards parasitoidism appears to be correlated also with a

capacity for the adult female to undergo physogastry, an immense swell-
ing of her body while feeding on a host, before reproduction, to accom-
modate nearly simultaneous development of many embryos, so that they
are even-aged in adulthood (see Figures 2-5). Hammen’s (1980) definition
of physogastry as “excessive enlargement of female opisthosoma after
fertilization” is inaccurate in several respects. First, enlargement is not
excessive, but extensive. Second, enlargement is not necessarily limited to
the opisthosoma, but may include the hysterosoma (the part of the body
bearing legs I11 and IV) or the entire idiosoma, as in the tarsonemid genus
Zponemus.Third, physogastry may ensue with feeding on a host whether or
not copulation or fertilization has taken place, as indicated by unmated,
unfertilized physogastric females giving rise to an all-male generation of
offspring. As a parasitoid female needs only one host for herself to
reproduce and for her progeny to develop to adulthood, she can afford to
lose mobility, enlarge, and make greatest use of the nutrient resource of her
host. In contrast, predatory females must remain mobile to capture other
prey. Physogastry has also been documented in some fungivorous taxa of
Tarsonemina, including Dolichocybidae (Rack, 1967; Kantaratanakul e f
al., 1989), Trochometridiidae (Cross and Bohart, 1978) and Siteroptidae
(Krczal, 1959; Rack, 1972; Suski, 1973; Zou et al., 1993). These families
have been derived earlier than those in which parasitoidism is known to
occur (Microdispidae, Pyemotidae, Tarsonemidae), and this argues both
for physogastry as a preliminary step towards parasitoidism and for
parasitoidism to have arisen from fungivorous, rather than predaceous,
ancestors. In fact, physogastry has not been documented for any predatory
taxa in the Acari.
4. DISTRIBUTION OF PHYSOGASTRY, PARASITISM AND

PARASlTOlDlSM AMONG TARSONEMINA
The placement of families and arrangement of superfamilies of Tarsone-

mina (see examples of habitus in Figures 2 to 17) presented in the follow-
ing discussion differ considerably from previous classifications presented
HETEROSTIGMATA
DOUCHOCYBOIDEA
TARSONEMINA TROCEOUETRIDIOIDEA
Figure I Proposed cladogram of the heterostigmatic superfamilies of mites

recognized in this chapter (after Lindquist, 1976, 1986; Lindquist er al., 1990;
M. Kalizsewski, unpublished data).
by other authors, e.g. Mahunka (1970), Lindquist (1973, 1986) and Krantz
(1978), and this requires some explanation here. Krantz’s (1978) manual
largely followed Mahunka’s (1970) classification of the Tarsonemina in
having three superfamilies: Pyemotoidea with four families, Dolichocybi-
dae, Caraboacaridae, Pyemotidae, and Acarophenacidae; Pygmephoroidea
with three families, Scutacaridae, Pygmephoridae (including Trocho-
metridiinae and Siteroptinae as subfamilies), and Microdispidae; and
Tarsonemoidea with two families (Tarsonemidae and Podapolipidae).
Lindquist ( 1986) recognized that Trochometridiidae and Dolichocybidae
had to be accommodated in separate superfamilies to account for their
more ancestral, sequential relationships as out-groups to the other super-
families. More recently, the senior author’s continuing cladistic studies
indicated additional classificatory changes, including recognition of
Scutacaroidea (with families Scutacaridae and Microdispidae) as a sister
superfamily separate from Pygmephoroidea (with Pygmephoridae and
Siteroptidae as separate families) and transfer of Caraboacaridae from
Pyemotoidea to Trochometridioidea, hich also includes Athyreacaridae
‘$I
as described by Lindquist et al. (1990 , This classification is summarized
by the dendrogram in Figure 1. The phylogenetic rationale for most of its
branching sequences, based on apomorphies, is presented by Lindquist
(1976, 1986); however, the basis for the sister branches Scutacaroidea
and Pygmephoroidea has yet to be presented, and that for the branch
Trochometridioidea needs further refinement.
4.1. Superfamily Dolichocyboidea
Little is known about the ways of life of species of the one family in this
group, which is recognized as the earliest derivative, or most “primitive”,
among the described families of Tarsonemina (Lindquist, 1986). Never-
theless, these mites are highly specialized morphologically, and also
manifest several specialized biological attributes (Figures 3-6). They
appear to be associated with subsocial and social insects, particularly
beetles and ants, on which they are phoretic as adult females. Rack
(1967) documented that adult females of Dofichocybe hippocastani Rack
are fungivorou’s,undergo extensive hysterosomal phy sogastry during feed-
ing, and give rise to adult male and female progeny. Physogastric females
produced from 30 to 200 progeny each, with the sex ratio strongly biased
towards females; 70% to 96% of progeny were females. Most progeny
included three to nine males. Mating occurred either inside the dying
maternal body sphere or immediately after birth. Birth appeared to be a
mass emergence of several to many females and males together through
ruptures of the maternal body sphere, rather than sequentially through a
birth canal. Multiple matings of new adult females with the same or
different males were observed. Multiple matings with different males
may reflect the lesser degree of sexual dimorphism in Dolichocybidae,
with concomitant lack of special leg modification in the males for selec-
tive precopulatory transport of females, in contrast to those found to some
extent in all other superfamilies of Tarsonemina. When Rack (1973)
described Dolichomotes crossi Rack, she noted that one female was gravid
and strongly physogastric, containing about 47 progeny. Feeding habits
were not confirmed but the sample of mites was collected under bark of
walnut trees (Jugfans),where fungal growth probably occurred. Similarly,
Dofichocybe keiferi Krantz was collected from dying limbs of maple trees
(Acer),where there was some fungal growth and some of the female mites
were noted as “swollen” (Krantz, 1957). The phenomenon of adult female
dimorphism has been determined for one member of this family, in the
genus Formicomotes, by Magowski (1988). The biological aspects of
fungivory, extensive opisthosomal physogastry, high fecundity (105-355
progeny per female), and strongly female-biased sex ratio (c. 98% female)
of Formicomotes heteromorphus Magowski, as reported by Kantaratanakul
e f al. (1989), are similar to those mentioned above for B.hippocastani. In
general, dolichocybid mites do not appear to be parasites of their hosts.
However, their adult females, basically fungivorous, physogastric, and
phoretic, may in some cases have undergone adaptations in the direction
of facultative parasitoidism, as discussed next for Trochometridium.
mum
Figure 2 Heterocheylus sp., dorsum of the female (after Lindquist, 1986).

Figures 3-6 Dolichocybe hippocastani (Dolichocyboidea). 3: Female, dorsum
and ventrum; 4: male, dorsum and ventrum; 5: physogastric female; 6: gravid
physogastric female and detail of an embryo. (After Rack, 1967.)
Figure 7 Trochometridium sp. (Trochometridioidea), ventral view of a female.
(After Lindquist, 1986.)
4.2. Superfamily Trochometridioidea
Presently, only three genera of trochometridioid mites (Caraboacarus,

Trochometn'diwm, Athyreacarus) have been described (Cross, 1965;
Lindquist et al., 1990), each now placed in a separate family (Figure 7).
Members of the genus Caraboacarus are found under the elytra of various
carabid beetles, and in some cases have been observed with their chelicerae
inserted into the veins of the host's hind wing (Nickel and Elzinga, 1969).
Based upon these observations, as well as the structure of the gnathosoma,
Cross (1965) and Nickel and Elzinga (1969) suggested that these mites
were external parasites. This is doubtful, as the male and other instars of
Caraboacarus have not been found on their hosts. Parasitoidism, however,
remains a possibility.
The life cycle of Throchometridium tribulatum Cross, which is the only
species of the genus extensively studied, is relatively complicated and
closely correlated with the life history of ground-nesting bees (Cross and
Bohart, 1969, 1978). The adult female mite is phoretic on the female alkali
bee, Nomia melanderi Cockerell. The maternal bee provides each brood
cell with pollen to feed a single offspring, lays an egg, and then closes the
cell. The mite disembarks during this process and becomes sealed within
the bee cell. She then apparently discharges the fungal spores that have
been carried in her sporothecae (Lindquist, 1985), and these germinate
mycelial growth. It is possible that the mite kills the bee egg or larva
soon after the cell is sealed, as living bee larvae have not been found in
cells containing mites. If the bee larva survived, it could feed on the pollen,
which is the nutritive substrate of the fungus, or even impede the dev-
elopment of mycelium by the production of antibiotic substances. The
female mite feeds on the mycelium, undergoes physogastry, and repro-
duces. Her larval offspring are non-feeding, as are the few emergent adult
males. It is doubtful whether new adult female mites feed immediately
after emergence, as they do not undergo phsyogastry then. After mating,
adult females of the new generation escape from the cell and seek female
bees to which to attach. The female of T. tribulatum reproduces only in bee
cells which do not contain a living larva, so the effect is comparable to that
of a parasitoid, although the mite does not consume the host. This may also
be the case with adult female mites of the recently described family
Athyreacaridae associated with scarab beetles. Although feeding habits
and other aspects of their reproductive behavior are unknown, their
cheliceral and pharyngeal structures closely resemble those of Trochome-
tridiidae, and extensive areas of highly plicated soft cuticle indicate their
physogastric capability (Lindquist et af., 1990). These may well be exam-
ples of an intermediate evolutionary step towards full parasitoidism from a
lineage of fungivores that was already capable of physogastry. As noted
above, fungivory with physogastry has been documented for adult females
in the Dolichocybidae, the earliest derived family of Tarsonemina which is
known.
4.3. Superfamily Scutacaroidea
Most members of the family Scutacaridae (Figures 8-10) are free-living or

animal-associated forms without indication of parasitoidism or parasitism
(Athias, 1972; Ebermann, 1991). Many Scutacaridae are associated with
social insects. Many, perhaps most, species of the group are phoretic, and
dimorphism among the adult females is frequent (Ebermann, 1991). The
presence of female dimorphism is fully correlated with phoresy in this
family. However, some species that are phoretic have been shown to have
only one female morph (Ebermann, 1991). Whether this indicates the loss
of the non-phoretic morph, i.e. a secondary suppression of female dimor-
phism, is uncertain; however, species with monomorphic females are found
in several genera, all of which have other species with dimorphic females.
A limited amount of swelling of the body of gravid females has been noted
in a few fungivorous scutacarids, and referred to as physogastry by Binns
(198 1). This is not equivalent to the well-developed physogastry, with
development of many even-aged progeny, discussed for other families
herein; however, it may represent a step in that direction.
Parasitoid association with insects has evolved at least twice in the
family Microdispidae. The genera Glyphidomastax and Perperipes are
associated with army ants, and most probably feed on their larvae and/or
eggs. They also use adult ants for phoretic transport. We know only
phoretic females of these genera and lack information about their life
histories. However, the unusual structure of their gnathosoma supports
the hypothesis of Cross (1965) concerning their parasitoid association
with army ants. Their palpi are very long, terminating in two elongate,
bladelike teeth, which are probably used for piercing the ants’ cuticle or the
chorion of their eggs. These palpi are analogous to the chelicerae of other
parasitic or parasitoid Tarsonemina. Glyphidomastax and Perperipes most
probably arose from free-living, fungivorous forms, as the gnathosomal
structure of their sister group, Myrmecodispus, indicates that it is probably
fungivorous. The only known species of the microdispid genus Peponocara
most probably is a parasitoid of the polyphagine roach, Arenivaga apacha
(Saussure) (Cross, 1965).
8
lOOp
\ 9
I
'7'
I
T
Figures 8-1 0 Scutacarus palustris (Scutacaroidea). 8: Adult female dorsal

(upper left) and ventral (lower right); 9: tarsus of leg I, note the enlarged claw
adapted for phoresy; 10: leg IV. (After Rack, 1966.)
Figure 1 1 Pygmephorus punctatus (Pygmephoroidea), adult female, dorsal (left)
and ventral (right). (After Rack, 1975.)
Figures 12-1 3 Luciaphorus perniciosus (Pygmephoroidea). 12: Female, dorsal;
13: Male, dorsal. (After Rack, 1983.)
4.4. Superfamily Pygmephoroidea
The two families of Pygmephoroidea (Siteroptidae and Pygmephoridae)

consist primarily of free-living species with phoretic or non-phoretic
female forms, without indication of parasitoidism or parasitism (Figure
11). In the Siteroptidae, species of Siteroptes and Pediculaster, often
associated with bark beetles and fungus-inhabiting flies, are free-living
fungivores often with phoretic and non-phoretic female dimorphism
(Smiley and Moser, 1976; Martin, 1978; Cross and Kaliszewski, 1988).
Similar observations by Kosir (1975) and other authors on fungivorous
species of “Pygmephorus”, especially “ P ” ..mesembrinae, refer in whole
or in part to siteroptids (Cross and Kaliszewski, 1988). Adult females of
some species undergo immense hysterosomal physogastry while feeding,
with resultant high fecundity. In Siteroptes cerealium (Kirchner) (=S.
graminum Reuter), for example, an average of 100-200, but up to 500,
progeny per female may be produced (Cooper, 1937; Krczal, 1959), of
which about 5% are male. The progeny pass through an active larval instar,
and usually attain adulthood within the maternal hysterosomal sphere,
where copulation often takes place before birth. A mass birth usually
occurs with rupture of the maternal body and her death. The life history
of Luciaphorus perniciosus Rack, 1983 (Figures 12-13) - which was
reported under the name L. hauseri Mahunka (1981) by Kantaratanakul
and Jitrar (1984) and by Kaliszewski and Wrensch (1993) and under the
name L. auriculuriae Gao et al. (1 990) by Zou et al. (1993) - is similar to
that of species of Siteroptes. The female is fungivorous and undergoes
extensive physogastry, with development of her progeny to adulthood, and
sometimes copulation, occurring within her body. Adult female dimor-
phism and phoresy are not recorded for the two species of Luciaphorus
described; however, the enlargement of leg I, which has a thick, grasping
claw, indicates phoretic behavior in the one form of female that is known
for each of these species.
In the family Pygmephoridae, adult females of various genera use insects
or mammals for phoretic dispersal. Those in genera associated with insects
are generally free-living, fungivorous commensals of their associates (Rack
and Eickwort, 1980; Ebermann and Rack, 1982), and female dimorphism is
known or thought to occur in many of them (Rack and Kaliszewski, 1985).
Members of the genus Pygmephorus commonly occur in the nests of
small mammals, and are phoretic in the fur of the mammals or on insects
frequenting their nests. Although nearly 50 species have been described as
phoretic adult females, little is known of their biologies. Curiously, males
of only two species are known, data on reproduction and post-embryonic
development are lacking, and non-phoretic females have not been reported
(Dastych et al., 1992).
4.5. Superfamily Pyemotoidea
Two families of Pyemotoidea (Pyemotidae and Acarophenacidae) are

parasitoid on pre-imaginal Lepidoptera, Homoptera, Coleoptera, Diptera
and Hymenoptera (Krczal, 1959; Rack, 1959; Cross and Krantz, 1964;
Cross, 1965; Cross and Moser, 1971). Adaptation to the parasitoid way
of life may be seen in gnathosomal modifications, namely enlarged
cheliceral stylets, reduced palpi, and a well developed pharyngeal pump
(Figure 14). High fecundity is achieved by a relatively short life cycle
(suppression of active larval instar) and development of many eggs at
approximately the same time, facilitated by extreme physogastry of the
mother on a single host. Physogastry appears to be an advantageous trait in
producing numerous, even-aged, mature progeny, ready to mate and to
attach to a carrier. In adult female Pyemotidae, physogastry may be very
extended, yet limited to the opisthosoma; in Acarophenacidae, it is less
extended but affects the entire idiosoma.
The biology of most pyemotids remains unknown but, based on studies
of a few species, Cross and Moser (1971) and Moser et a f . (1987a)
concluded that the 15 then-known species of the genus Pyemotes are all
parasitoids (referred to as “parasites” in these papers). The genus naturally
divides into two distinct groups, the P . scofyti and the P . ventricosus
groups (Cross and Moser, 1975; Moser et al., 1987a). Species of the P .
scofyti group are stenoxenous, and usually associated with bark beetles.
Their females are di- or polymorphic, apparently non-venomous, and feed
on the eggs, larvae and pupae, but usually not adults, of their hosts. Species
of the P . ventricosus group are euryxenous associates of insects in a wide
variety of habitats; their females are monomorphic, highly venomous, and
attack all instars of their hosts.
In the Pyemotes scolyti group, for example, P . parviscolyti Cross and
Moser rides both sexes of a pine bark beetle, Pityophthorus bisufcatus
(Eichhoff), but no other pine bark insects (Moser et al., 1971). In the field,
the first generation issued from a phoretic maternal mite attacks eggs, the
second generation feeds on larvae, and so on until imrnatures become
unavailable; adult beetles are not attacked. Another species in this group,
P. scofyti (Oudemans), is possibly an assemblage of cryptic species. Moser
and Roton (1972) showed that the reproductive compatibility between
morphologically similar populations of P . scolyti associated with Scolytus
muftistriatus Marsham originating from Europe, and those phoretic on
Scolytus ventralis LeConte from North America, was incomplete. This
separation seems to be due to ecological barriers between their phoretic
hosts, with S. multistriatus on elms (Ulmus spp.) and S . ventralis on
conifers (Abies, Picea and Tsuga spp.)
In contrast, species of the P. ventricosus group have a wide host range,
14 15
17
Figure 14 Acarophenav nidicolus (Pyemotoidea), ventral view of female. (After

Krantz, 1978.)
Figure 15 Tarsonemus sp. (Tarsonemoidea), dorsal view of female. (After
Krantz, 1978.)
Figure 16 Tursonemoides sp. (Tarsonemoidea), ventral view of female. (After
Krantz, 1978.)
Figure 17 Acarapis externus (Tarsonemoidea), adult female, ventral aspect on
left side, dorsal view on right. (After Lindquist, 1986.)
and are frequently encountered in insect cultures and stored foods. The
ubiquitous and euryxenous Pyemotes tritici (Lagr&ze-Fossat and
MontagnC) is a significant mortality factor for stored product insects
(Tawfik et al., 1981). It has been used (under the name P . helfsi) in
biological control of insects such as the pink bollworm, Pectinophora
gossypiellu (Saunders), in Egypt (Tawfik and Awadallah, 1971), and it
has even been introduced for biological control of the imported fire ant,
Solenopsis invicta Buren, in the United States with inconsistent success
(Bruce and LeCato, 1980; Thorvilson and Phillips, 1987). All attacked prey
die immediately, paralysed by neuromuscular toxins, and the mite attacks
all instars of the host. Thus, this form should be considered as a micro-
predator, the mite being much smaller than its prey. This species manifests
one of the highest reproductive rates known for any animal (Bruce and
Wrensch, 1990), with such attributes as a short life cycle (4-7 days), no
intermediate host, numerous progeny (average 254, maximum number
observed 355), all development of progeny occurring within the mother’s
engorged body, and all offspring born as sexually mature adults. Females
represent 95-99% of the population, and they mate immediately after birth
and seek a host. Populations are easily cultured in the laboratory, and the
species is routinely commercialized (Moser et al., 1987a, b). The bites of
females in the P . ventricosus group, e.g. P . helfsi (Oudemans) (syn. P .
zwoelferi Krczal) and P . tritici, are so toxic that they cause a pruriginous
dermatitis, called “grain itch”, of granary workers (Krczal, 1959; Moser,
1975; Le Fichoux et al., 1980). Another member of the P . ventricosus
group, Pyemotes barbara Moser, Smiley & Otvos, is a parasitoid of the
Douglas-fir cone moth, Barbara colfuxiana (Kearfott). Its females feed
naturally on the pupa of this host, but they readily attacked immature
instars of other forest insects offered in the laboratory, and were suggested
as candidates for the biological control of other cone and seed insects
(Moser et al., 1987a, b).
The kind of ovoviviparous development occurring in species of Pye-
motes results in only adult females and males being directly exposed to
external environmental factors. Further, the sex ratio is highly biased;
males usually represent only 1-20% of the progeny, which still suffices
for nearly all the emerging females to be fertilized. These traits are
advantageous in that adults are the only “units of selection”, the maternal
genome is exposed in haploid condition in her sons, and the female diploid
genome is optimized both in number of individuals, and dispersants
(Maynard Smith, 1989; Bruce and Wrensch, 1990; Kaliszewski and
Wrensch, 1993). During the birth of adult progeny from the maternal
body, male(s) issue first, which facilitates their mating with the females
immediately, or shortly after, they are born. In some species, the male
assists and accelerates the birth process of his sisters, but apparently no
assistance is given in others (Moser, 1981). Assisted births of females take

from 0.3 to 3 minutes, whereas unassisted births commonly take about 3-
10 minutes, although the first-born may take several hours (Krczal, 1959;
Moser et al., 1971, 1978; Bruce and Wrensch, 1990). Single males mate
with an average of about 20-50 females each; occasionally, as many
matings as 57 for a male of P . tritici, 58 for P . dryas (Vitzthum), 78 for
P . herjsi (Oudemans), 131 for P . parviscolyti, and 258 for P. scolyti have
been noted, depending in part on male longevity (Bruce and Wrensch,
1990; Moser et al., 1978; Herfs, 1926; Moser et al., 1971; and Krczal
1959, respectively). When several maternal mites are clustered on one host,
two or more males usually aggregate and often move from mother to
mother, a mechanism which may reduce inbreeding risks (Moser et al.,
1971). Notably, copulation, as distinct from fertilization, induces searching
behavior by young adult female mites for a carrier. By crossing virgin
females of P . parviscolyti with males of P . ?ventricosus and vice versa,
Moser et al. (1971) triggered seeking behavior in unfertilized females. The
progeny were always all males, of the same species as the mother.
Host/phoretic and host/parasitoid specificities are important limiting
factors in considering species of parasitoid mites for potential use in
biological control (Athias-Binche, 1990). For example, cultures of Pye-
mores dryas, a European species that attacks bark beetles, were brought to
Louisiana in order to evaluate its potential as a control agent of the
southern pine beetle, Dendroctonus frontalis Zimmermann. In laboratory
cultures, the females fed and developed only on late larvae and pupae of D .
frontalis, but the sons progressively became feeble after 2-50 generations.
More important, despite this mite’s having a large range of hosts in Europe,
it did not attach to American beetle species, whether D . frontalis or other
sympatric insects (Moser et al., 1978). Thus, dispersal was impossible, and
the parasitoid population was doomed to disappear, in contrast to P . tritici,
which can use a great variety of carriers.
In Acarophenacidae, the sister family of Pyemotidae, adult females are
parasitoids of insect eggs (Figure 14). In the genera Adactylidium and
Acarophenax, an inseminated female usually produces only one male. In
one species of Adactylidium, the male apparently fertilizes his sisters inside
the engorged body of their physogastric mother, before they are born (El
Badry and Tawfik, 1966). In this special case, inbreeding seems to be
unavoidable, and genetic drift by founder effects should lead to strict
monoxenous specificity, as noted by Lindquist (1969b) for egg parasitoids
of the tarsonemid genus Zponemus. In species of Acarophenax, the male
fertilizes his sisters inside the mother’s body, just before their birth, or
outside, immediately thereafter (Newstead and Duvall, 1918; Rack, 1959).
According to Maynard-Smith (1989), a female should produce only one
son, haploid with her genome, if this is advantageous in maximizing the
selective fitness of her genome on one hand yet transmitting it through as

many of her dispersant female offspring as possible on the other hand. As
pointed out by Bruce and Wrensch (1990), however, there should be a
sufficient number of males produced to ensure that nearly all females are
mated before dispersal. In the cases of the pyemotid species discussed
above, several males are needed from each maternal female, because of
her high fecundity. But in the cases of the acarophenacid species discussed
here, one male evidently suffices for each maternal female, because of her
less extended idiosomal physogastry and moderate fecundity (about an
order of magnitude less than in some pyemotids), e.g. 4-15 progeny per
female in Acarophenux tribolii Newstead and Duvall (1918), 3-27 in A.
dermesridurum Rack (1959), and 6-9 in Aductylidium sp. (El Badry and
Tawfik, 1966). A . muhunkui Steinkraus and Cross (1993) differs in produ-
cing more progeny (8-56, mean 29) and occasionally more males (two to
four instead of one, in 40% of the females examined); however, one male
evidently still suffices for any number of female offspring produced
(Steinkraus and Cross, 1993).
The sex ratios in these two families indicate how readily natural selec-
tion can engender the most favorable female-biased sex ratio possible in
haplo-diploid organisms, especially in parasitoids such as these. In terms of
the cost of dispersal, the production of as few males as are needed to achieve
fertilization of nearly all females is advantageous to the economy of the
reproduction budget in Tarsonemina, because sons are non-dispersant and
the offspring consists mostly of migrant, colonizing daughters. The sex
ratio can therefore be highly biased, and culminates in the production of
only one son, as in the Acarophenacidae.
Certain members of the genus Acurophenax are parasitoids of gramini-
vorous beetles of the genera Tribolium and Cryptolestes (Newstead and
Duvall, 1918; Cross and Krantz, 1964). Steinkraus and Cross (1993)
recently described a new species, A . muhunkui, attacking the eggs of the
lesser mealworm, Alphitobius diuperinus (Panzer), a serious cosmopolitan
poultry pest. Acurophenux meropsi Rakha and Kandeel (1983) was
described from the European bee eater in Egypt. However, as the bee
eater is an insectivorous bird living in cavities, an accidental transfer of
this mite from insects to the birds is suspected. All species of the genus
Purucurophenux appear to be parasitoids of beetles (Rack, 1959; Mahunka,
1975). Three species of Aductylidium are parasitoids of thrips (El Badry
and Tawfik, 1966).
4.6. Superfamily Tarsonemoidea
The family Tarsonemidae manifests a greater diversity of feeding habits

than in any other family of mites (Lindquist, 1986; Figures 15-17). Free-
living members of the group are fungivores and algivores, and many feed
on vascular plants. Plant-feeding Tarsonemidae usually adopt anemo-
chorous dispersal. However, the broad mite, Polyphagotarsonemus latus
(Banks), which infests crops in tropical regions, has been recorded several
times as phoretic on the whitefly, Bemisia tabaci Gennadius (Homoptera:
Aleyrodidae) (Flechtmann et al., 1990). This may represent a preliminary
step in evolution towards parasitism or pqasitoidism in plant-inhabiting
Tarsonemidae, particularly if the broad mite occasionally sucks body fluid
’ from its carriers, as claimed by Gupta and Chaudhry (1972) but doubted by
Lindquist (1986).
Within the family Tarsonemidae there are genera well documented as
parasitoids and parasites, as well as predators. Species of the genus
tponemus are exclusively parasitoids specifically associated with scolytid
beetles; 16 of the 18 species associated with pine-feeding ipine beetles in
North America are monospecific (Lindquist, 1964, 1969~).The size of the
unengorged females of most species is directly proportional to the size of
the adult host. This correlation possibly reflects a relationship between the
size of the mite and the size of the beetle egg, as the egg may function as a
limited food supply. Adult females of Zponemus feed on the beetles’ eggs,
and use young, emerging adult hosts for dispersal, phoretic on the elytral
declivity of the carrier. The young adult beetles mate after dispersal, then
build their galleries and lay their eggs. These activities induce detachment
of the mites, which disembark and seek host eggs in the galleries. A female
mite pierces an egg and feeds for about 2 days. It undergoes idiosomal
physogastry, lays 40-80 eggs, and dies. Lindquist and Bedard (1961) noted
that, when several mites engorge on a single egg, fewer eggs per female are
laid, or oviposition may be aborted if there is a high level of trophic
competition. This indicates again that a certain level of feeding and
physogastric engorgement, rather than copulation, is necessary before
oogenesis begins. Eggs are laid externally to the maternal mite’s physo-
gastric sphere. After hatching, the mite larvae are fully mobile with well-
developed legs. However, they remain clustered together and do not feed,
using instead the fatty yolk content of their body derived through the egg
from the mother’s reserves (Terry, 1966; Lindquist, 1969~).Male adults
emerge first, and usually represent about 5% of the progeny. It is important
to note that they remain around the mother’s body and adherent cluster of
progeny, and mate with newly emerging female adults. Thus, a few males
suffice to fertilize nearly all of the many females, which are usually their
sisters, a situation which leads to much inbreeding. Evidently, the
moderately high number of progeny per female requires more than the
minimal one son to ensure the mating of nearly all daughters, much as in
Pyemotidae rather than Acarophenacidae. The fertilized, unfed, young
females then seek a carrier. To synchronize with the host’s life cycle,
this unfed, pre-phoretic and phoretic stage of the mites may last from 2 to
4 weeks during the active season under temperate conditions, or up to 8
months in fall and over winter (Lindquist and Bedard, 1961; Balazy and
Kielczewski, 1965; Lindquist, 1969~).
Adult females of the genus Tursonemellu have been recorded as disper-
sants on adult chalcidoid wasps and undetermined beetles (Lindquist,
1986). Based on their spectacularly long cheliceral stylets, Lindquist
(1986) suggested that these mites may be parasitoids on the immature
instars of their host. However, there are unpublished data which indicate
a remarkable symbiosis between some of these mites, their agaonid wasp
carriers, and host fig fruit. The pollinating agaonid wasp females carry the
mites through the ostiole into the fig fruit, where the mites initiate gall
formation by some of the fig ovules on which they feed in dense
aggregations (personal correspondence between S. G. Compton and E.
E. Lindquist, 1986-1 989; Compton, 1993).
There are few documented examples of predation among tarsonemid
mites. Species of Acaronemus prey on eggs of tetranychid and tenuipalpid
mites on the foliage of evergreen trees (Smiley and Landwehr, 1976;
Lindquist and Smiley, 1978). Evidence that some members of Dendrop-
tus, the sister-genus of Acaronemus, may prey on gall-producing eriophyid
mites, or on eggs of arboreal tydeid mites, was reviewed by Lindquist
(1986). These are relatively highly derivative genera, and there appears to
be no evidence for their representing an intermediate step between
fungivory and parasitoidism or parasitism in the family.
All instars of Tursonemus (Chuetotarsonemus) dispur Beer live as
symbionts beneath the elytra of the passalid beetle, Popilius disjunctus
(Illiger) (Beer, 1954; Lindquist, 1986). As their cheliceral stylets are not
enlarged, these mites are perhaps paraphagic, though parasitism should not
be dismissed.
The adult female of Pseuducurupis indoupis (Lindquist) was described
as a phoront on Apis cerunu Fabricius (= A. indicu Fabricius) in India
(Lindquist, 1968, 1986), and we have a similar record of it from Thailand.
Its body shape is similar to that of females of the parasitic tarsonemid
genus Acurupis, and it secludes itself in the same areas on the bee as does
Acurupis externus Morgenthaler (Figure 17), i.e., the cervix and posterior
head near the posterior tentorial pits. In contrast to species of Acurupis, the
presence of only the adult female on the bee, and the short cheliceral stylets
and small pharynx of Pseuducurupis, do not indicate parasitism. Further,
this is a relatively highly derived genus, which could not represent an
intermediate step towards the parasitism evolved in the much earlier-

derived lineage leading to Acurupis (see Lindquist, 1986). The small,
weakly musculated pharynx of adult female Pseuducurupis is also not
indicative of parasitoidism, and Lindquist (1968, 1986) suggested that it
may be adapted to living as a pollen-feeding commensal with bees.
Mites of the genus Acurupis and all genera of the tarsonemid tribe
Coreitarsonemini are highly host-specific parasites of adult insects.
Although highly specialized morphologically for a parasitic way of life,
these taxa represent a relatively early derivative, or “primitive”, lineage of
Tarsonemidae (Kaliszewski, 1984; Lindquist, 1986). Species of Acurupis
live as parasites only on adult honey bees (Apis spp.). Parasitism by
Acurupis woodi (Renni) in the tracheal system of Apis melliferu Linnaeus
apparently hastens the physiological ageing of the bees. The mites injure
hosts in two ways: they pierce the wall of the tracheae to feed on haemo-
lymph, which also causes degeneration of tissues around the feeding site,
and their increasing numbers apparently reduce the exchange of air through
tracheae, which increasingly hinders respiration. It has frequently been
assumed that such tracheal infestation impairs the bee’s ability to fly,
and that the bee eventually dies of starvation or suffocation. This
pathogenicity is commonly known as “acarine disease” or “acario-
sis” (Lindquist, 1986). Two or three other species of Acurupis have been
described, and documented as parasites on the external surfaces of honey
bees (Morgenthaler, 1934; Bailey, 1963). Parasitism by these species does
not appear to be debilitating (Eckert, 1961).
The tribe Coreitarsonemini includes the genus Coreitarsonemus, with 2 1
described species or subspecies, and the monobasic genera Amcortursone-
mus and Asiocortursonemus. Larvae and male and female adults have been
found only on coreid bugs representing at least 24 genera, and appear to
infest only the atria of the odoriferous thoracic glands of their hosts (Fain,
1970, 1971, 1978, 1980a, b). Patterns of host specificity are not clear, but at
least one species, C. unoplocnemis Fain, has been recorded from hosts
representing 11 species and 10 genera of Coreidae (Fain, 1978). Based on a
greenish coloring in the body of some of these mites, Fain (1970) suggested
that they may be phytophagous, using the host for refuge and dispersal to
plants visited by the insects. However, Lindquist (1986) indicated that
these mites are probably parasitic, noting that the haemolymph of many
insects is greenish, and that tarsonemids using insects primarily for dis-
persal do so only as adult females, and not in such a specialized site. Also,
the presence of great amounts of finely plicated soft cuticle between the
sclerotized plates on the body indicates a physogastric capability in adult
females of this tribe, and this also correlates with parasitic or parasitoid
feeding habits, rather than phytophagy, in the Tarsonemina (Lindquist,
1986).
As with many other mite lineages which have evolved towards parasit-
ism (Parasitengona (mostly the Hydracarina) in Actinedida, Psoroptides in
Acaridida, Dermanyssoidea in Gamasida), one notes a tendency to lose the
phoretic means of dispersal as all mites remain in all stages on the host
(Athias-Binche, 1991; Athias-Binche and Morand, 1993). This evolution-
ary trend is also encountered in Tarsonemoidea: Acarapis and the Corei-
tarsonemini in the family Tarsonemidae, and all taxa in the sister family
Podapolipidae as parasites infest a new host directly from another host
(Lindquist, 1986). Movement between hosts is primarily by adult female
mites in Acarapis (and probably in Coreitarsonemini), but there is a trend
for this behavior to shift from the adult female to the larval female in
Podapolipidae.
Mites of the family Podapolipidae are exclusively internal or external
parasites of insects. Podapolipids occur under elytra and on vaginal mem-
branes of beetles, in the trachea and air sacs of orthopterans, and externally
on grasshoppers and cockroaches. Most podapolipid genera exhibit reduc-
tion of the legs and other morphological structures. However, although
the adult females show the greatest trend towards leg reduction and
becoming a reproductive sac, the larvae consistently retain three pairs
of well-developed legs, evidently indicating the need for some movement
for transfer between hosts. This phenomenon is evidently connected with
the completely parasitic way of life throughout the entire family (Regenfuss,
1968; Lindquist, 1986).
However, members of the earliest derived genus known, Chrysomelobia,
which are subelytral parasites of leaf beetles in Europe and North America,
retain four pairs of legs in the adult male and female. Most species of
Podapolipus, Podapolipoides, Peripolipus, Panesthipolipus and Bakerpo-
lipus are external parasites of Orthoptera (Husband, 1984). Species of
Rhynchopolipus occur on weevils (Husband, 1984). Species of Locusta-
carus occur in respiratory systems of locusts and bees, apparently piercing
tracheal walls to feed (Wehrle and Welch, 1925; Husband and Sinha,
1970). Members of the genera Tarsopolipus, Eutarsopolipus, Tetrapoli-
pus, Coccipolipus, Dorsipes, Chrysomelobia, Dilopolipus, Stenopolipus,
Archipolipus, Regenopolipus, Stigmarcarus, and some species of Podapo-
lipus feed externally on a variety of beetles (Regenfuss, 1968; Feldman-
Musham and Havivi, 1972; Baker and Eickwort, 1975; Husband, 1984).
Some species of Ovacarus are also parasites of beetles, but appear to be
restricted to the vaginal membranes and oviduct sacs of their carabid hosts
(Stannard and Vaishampayan, 1971; Husband, 1984).
5. DISCUSSION
The Tarsonemina manifest various attributes that have acted in the course
of evolution in favor of a diversity of symbiotic ways of life. Their small
size no doubt has facilitated general or loose symbioses, especially
phoresy. Abbreviated post-embryonic development, arrhenotoky, and the
variety of feeding habits have contributed to the success of dispersal and
colonization of an appropriate host or habitat. Physogastry, which appears
frequent in the group, also has permitted an optimal production of even-
aged dispersants or infective stages. In fact, it seems that true, free-living,
edaphic forms, which would occupy the original ecological niche of mites,
are infrequent in the group. In many cases, the pygmephorids and tarso-
nemids sampled in soil by classic extraction techniques such as the
Berlese-Tullgren funnel may preferentially inhabit ground nests of
various organisms. Most of the Tarsonemina are associated with other
animals or are fungivores or plant feeders, and many of them are found
in above-ground layers.
The Tarsonemina, then, comprises a typically non-conservative lineage
of mites which has colonized largely non-edaphic trophic niches in the
course of its evolution and diversification. Not surprisingly, various of the
aforementioned characteristics, especially phoretic dispersal, may even-
tually lead repeatedly towards parasitic and parasitoid ways of life. Such
patterns are encountered in other groups of mites, such as the Derma-
nyssoidea (Radovsky, 1994), Hydracarina (Parasitengona) (Mitchell, 1967;
Wiggins et al., 1980), and Psoroptides (Houck and O’Connor, 1991).
However, the Tarsonemina constitutes the outstanding example among
the Acari of a lineage showing independently repeated, complete loss of
a true, free-living way of life in adaptation to parasitism and, uniquely,
parasitoidism.
Parasitoidism has evolved in earlier as well as more recently derived taxa
among the superfamilies, and even within the families, of Tarsonemina.
For example, it arose in the lineage ancestral to the entire superfamily
Pyemotoidea, yet it has arisen in just one known genus, Zponemus, in the
most recent derivative subfamily of Tarsonemidae. That evolution to
parasitoidism is, phylogenetically, a more recent phenomenon in Tarso-
nemidae than in Pyemotoidea is supported also by life history attributes.
Although physogastric, adult female Zponemus still retain the primitive
characteristics of ovipositing their eggs outside the maternal body, and
of the eggs hatching into active larvae. There may not have been sufficient
evolutionary time, yet, for retention of the eggs and reduction of the larvae
to a regressed stage within the physogastric mother, as in the Pyemotoidea.
Parasitism, in contrast with parasitoidism, appears to have evolved in
only one of the most recent derivative superfamilies of Tarsonemina, the

Tarsonemoidea. Parasitism evolved in the lineage ancestral to the entire
family Podapolipidae, as well as independently in the subfamily
Acarapinae in the Tarsonemidae. Nevertheless, we would not be surprised
to find parasitism in earlier derived taxa of Tarsonemina, and this is well
expressed in the Heterocheyloidea, a group of highly specialized parasites
of passalid beetles, yet derived earlier than the Tarsonemina, in the basal
lineage of Heterostigmata (Figures 1 and 2) (Lindquist and Kethley, 1975).
In view of the examples presented above, it appears that lineages in
Tarsonemina that have evolved towards parasitism are distinct from those
that have evolved towards parasitoidism. Parasitoidism does not appear to
lead to parasitism. Instead, each is an orthoselective end in itself, although
parasitism may continue to lead towards commensalism in some cases. The
evolutionary schism between trends towards parasitism and those toward
parasitoidism seems to be the acquisition of well-developed physogastry
and a capability to produce many even-aged progeny. Once this step is
reached, the evolutionary pathway to parasitism has been d pa s c-ed.
If these hypotheses of distinction between parasitism and parasitoidism
hold, some predictions can be made for taxa of Tarsonemina (and, more
broadly, Heterostigmata) associated with other species of arthropod hosts.
1. When all stages of a taxon are encountered on one host individual, the
taxon may be a parasite, or a commensalistic or mutualistic associate,
but it is not a parasitoid. When only the adult female of a taxon is found
on an adult host, the taxon may be a parasitoid, or a commensalistic or
mutualistic associate, or simply a phoretic, but it is not a parasite.
2. Adult female parasites are expected to undergo some engorgement
while feeding and fecund, but to generate and produce one or few
eggs at a time, sequentially over an extended time span (one to several
weeks). Adult female parasitoids are expected to undergo extended
physogastry while feeding and fecund, so as to generate and produce
a large number of even-aged progeny over a short time span (1-3 days).
3. The larva is expected to remain an active, feeding instar in parasitic
taxa, but to become inactive and non-feeding in parasitoid taxa. (In
Tarsonemidae, the nearly immobile larvae, with vestigial legs I1 and 111,
in the parasitic genus Acurupis, and the fully developed larvae, capable
of movement, in the parasitoid genus Zponemus, may appear to contra-
dict this; however, the former remain fully active as feeding parasites,
whereas the latter remain clustered together and do not feed.)
4. The sex ratio is not expected to be highly skewed (i.e., > 80%) in favor
'of females in parasitic taxa, but it is predicted to be so in parasitoid
forms.
5 . Among parasitoid and other taxa capable of extensive physogastry, the
360 MAAEK KALISZEWSKI ET AL
sons in an all-haploid progeny produced by an unfertilized female

cannot mate back with their mother, in her distended physogastric and
deteriorating state, to produce a second, bisexual generation. In some
parasitic and other taxa not capable of physogastry, infestation or
colonization of a new host or habitat by a virgin female can result in
her producing a generation of haploid males, and this may possibly be
followed by mother-son mating to produce a second, bisexual
generation.
6. Although there may be a dispersal instar, there tends not to be a phoretic
instar inactive on the carrying host in parasitic taxa. There is such a
phoretic instar, the adult female, in pgasitoid taxa. However, adult
‘
females of the parasitic mite, Acarupis woodi, blur this distinction
somewhat during transfer from one host bee to another. They actively
emerge from the tracheal system to an external site on the old host,
where they become temporarily inactive while waiting to attach to
another bee coming into contact with their old host, at which time
they reactivate and transfer to the new host (Hirschfelder and Sachs,
1952).
7. Adult females are not expected to be dimorphic in parasitic taxa, but
they may or may not be so among phoretic taxa, including parasitoids. If
dimorphic, one of the female morphs is specialized behaviorally and
structurally for phoresy.
ACKNOWLEDGEMENT
Figures 2-1 7 have been modified and entirely redrawn from several authors
by M. J. Bodiou, draftswoman, CNRS.
REFERENCES
Athias, F. (1972). Scutacaridae de la savane de Lamto (CGte d’Ivoire). 1. DonnCes

quantitatives sur le peuplement, liste des esphces de Scutacarinae avec
description de trois esphces nouvelles. Acarologia 14, 638-656.
Athias-Binche, F. (1984). La phorBsie chez les Acariens Uropodides (Anactino-
triches), une stratBgie Bcologique originale. Acta Oecologica, Oecologia
Generalis 5, 119-133.
Athias-Binche, F. (1990). Sur le concept de symbiose. L’exemple de la phordsie
chez les acariens et son Bvolution vers le parasitisme ou le mutualisme. Bulletin
de la Socittt Zoologique de France 115, 77-98.
Athias-Binche, F. (1991). Evolutionary ecology of dispersal in mites. Ecology and
PARASITISM AND PARASITOIDISM IN TARSONEMINA 361
evolution of phoresy in mites. In “Modem Acarology” (F. Dusbabek and V.

Bukva, eds), pp. 27-41. SPB, The HagueIAcademia, Praha.
Athias-Binche, F. (1994). Is the initiation of dispersal genetically fixed? The case
of phoretic mites. In “2nd Symposium, European Association of Acarologists
(EURAAC), Krynica, Poland, September 1992”. pp. 1-12.
Athias-Binche, F. and Morand S. (1993). From phoresy to parasitism: the example
of mites and nematodes. Research and Reviews in Parasitology 53, 73-79.
Athias-Binche, F., Schwarz, H. H. and Meierhofer, I. (1993). Phoretic association
of Neoseius novus (Ouds, 1902) with Nicrophorus spp., a case of sympatric
speciation? (Acari: Uropodina; Coleoptera: Silphidae). International Journal of
Acarology 19, 75-86.
Bailey, L. (1963). “Infectious Diseases of the Honey-bee.’’ Land Books, London.
Baker, T. C. and Eickwort, G. C. (1975). Development and bionomics of Chryso-
melobia labidomerae (Acari: Tarsonemina: Podapolipidae), a parasite of the
milkweed leaf beetle (Coleoptera: Chrysomelidae). Canadian Entomologist
107, 627-638.
Balazy, S. and Kielczewski, B. (1965). Tarsonemoides gaebleri Schaarschmidt
(Acar. Tarsonemidae) jajozerny roztocz w zerowiskach kornika drukarza Ips
typographus (L.). Polskie Pismo Entomologi B 37/38, 7-18.
Beer, R. E. (1954). A revision of the Tarsonemidae of the Western Hemisphere
(Order Acarina). University of Kansas Scientifrc Bulletin 36, 109 1-1387.
Binns, E. S. (1974). Notes on the biology of Arctoseius cetratus (Sellnick)
(Mesostigmata: Ascidae). Acarologia 16, 577-582.
Binns, E. S. (1981). Physogastry in Scutacarus baculitarsus Mahunka (Acarina,
Scutacaridae). Entomological Monthly Magazine 116, 157-159.
Bruce, W. A. and LeCato, G. L. (1980). Pyemotes tritici: a potential new agent for
biological control of the red imported fire ant, Solenopsis invicta (Acari:
Pyemotidae). International Journal of Acarology 6, 27 1-274.
Bruce, W. A. and Wrensch, D. L. (1990). Reproductive potential, sex ratio, and
mating efficiency of the straw itch mite (Acari: Pyemotidae). Journal of
Compton, S. F. (1993). One way to be a fig. African Entomology 1, 151-158.
Cooper, K. W. (1937). Reproductive behavior and haploid parthogenesis in the
grass mites, Pediculopsis graminum (Reut.) (Acarina, Tarsonemidae). Proceed-
ings of the National Academy of Sciences of the USA 2 3 , 4 1 4 .
Cross, E. W. (1965). The generic relationships of the family Pyemotidae (Acarina,
Trombidiformes). University of Kansas Scientifrc Bulletin 45, 29-275.
Cross, E. A. and Bohart, G. E. (1969). Phoretic behavior of four species of alkali
bee mites as influenced by season and host sex. Journal of Kansas Entomological
Society 42, 195-219.
Cross, E. A. and Bohart, G. E. (1978). Some observations of the habits and
distribution of Trochometridium Cross, 1965 (Acarina: Pyemotidae). Acarologia
20,286-293.
Cross, E. A. and Kaliszewski, M. J. (1988). The life history of a mushroom pest
mite, Pediculasterflechtmunni(Wicht) (Acari: Pygmephoroidea), with studies of
alternate morph formation. Environmental Entomology 17, 309-3 15.
Cross, E. A. and Krantz, G. W. (1964). Two new species of the genus Acarophenux
Newstead and Duvall 1918 (Acarina: Pyemotidae). Acarologia 6, 287-295.
Cross, E. A. and Moser, J. C. (1971). Taxonomy and biology of some Pyemotidae
(Acarina: Tarsonemoidea) inhabiting bark beetle galleries in North American
conifers. Acarologia 13, 47-64.
Cross, E. A. and Moser, J. C. (1975). A new dimorphic species of Pyemotes and

key to previously-described forms (Acarina: Tarsonemoidea). Annals of the
Cross, E. A., Moser, J. C. -and Rack, G. (1981). Some new forms of Pyemotes
(Acarina: Pyemotidae) from forest insects, with remarks on polymorphism.
International Journal of Acarology 7, 179-1 96.
Dastych, H., Rack, G. and Wilson, N. (1992). Notes on mites of the genus
Pygmephorus (Acari: Heterostigmata) associated with North American mam-
mals (Part 11). Mitteilungen aus dem Hamburger zoologischen Museum und
Institut 89, 141-156.
Dindal, D. L. (1975). Symbiosis: nomenclature and proposed classification.
Biologist 57, 129-142.
Ebermann, E. (1991). Das Phiinomen Polymorphismus in der Milbenfamilie
Scutacaridae (Acari, Heterostigmata, Tarsdnemina, Scutacaridae). Zoologia,
. Stuttgart 47, 3 (141), 1-79.
Ebermann, E. and Rack, G. (1982). Zur Biologie einer neuen myrmecophilen Art
der Gattung Petalomium (Acari, Pygmephoridae). Entomologische Mitteilungen
des zoologischen Museums von Hamburg 7( 115), 182-192.
Eckert, J. E. (1961). Acarapis mites of the honey bee, Apis mellifera Linnaeus.
Journal of Insect Pathology 3, 409-425.
Eickwort, G. C. (1975). A new species of Chrysomelobia (Acari: Tarsonemina:
Podapolipidae) from North America and the taxonomic position of the genus.
Canadian Entomologist 107, 613426.
El Badry, E. A. and Tawfik, M. S. F. (1966). Life cycle of the mite Adactylidium
sp. (Acarina: Pyemotidae), a predator of thrips eggs in the United Arab Republic.
Fain, A. (1970). Coreitarsonemusun nouveau genre d’acariens parasitant la glande
odorifkrante des Hkmipttres Coreidae (Tarsonemidae: Trombidiformes). Revue
de Zoologie et de Botanique africaine 82, 3 15-334.
Fain, A. (1971). Note sur la rkpartition gkographique des Coreitarsoneminae
parasite des Htmipttres corkides avec description de taxa nouveaux (Acarina:
Trombidiformes). Bulletin et Annales de la SociCtC Royale belge 107, 81-88.
Fain, A. (1978). Notes sur les Coreitarsoneminae (Acari, Prostigmata, Tarsone-
midae) parasites de la glande odoriftrante d’Hemiptbres Coreidae en Afrique
Centrale. Revue de Zoologie Africaine 92, 959-970.
Fain, A. ( 1980a). Les Coreitarsoneminae (Prostigmata, Tarsonemidae) parasites de
la glande odoriftrante d’Htmipttres de la famille Coreidae. I. Esptces d’Asie et
d’Australie. Acarologia 21, 279-290.
Fain, A. (1980b). Les Coreitarsoneminae (Prostigmata, Tarsonemidae) parasites de
.la glande odorifkrante d’Htmiptbres de la famille Coreidae. 11. Esptces
amtricaines. Acarologia 21, 346-360.
Farish, J. D. and Axtell, R. C. (1971). Phoresy redefined and examined in
Macrocheles muscaedomesticae (Acarina: Macrochelidae). Acarologia 13, 18-
29.
Fashing, N. J. (1975). Life history and general biology of Naiadacarus arboricola
Fashing, a mite inhabiting water-filled treeholes. Journal of Natural History 9,
4 13-424.
Feldman-Musham, B. and Havivi, V. (1972). Two new species of the genus
Podapolipus (Podapolipidae. Acarina), redescription of P . aharoni Hirst, 1921
and some notes on the genus. Acarologia 4, 657-674.
Flechtmann, C. H. W., Guerrero, J. M., Arroyave, J. A. and Constantino, L. M.
(1990). A little known mode of dispersal of Polyphagotarsonemus latus (Banks).
International Journal of Acarology 16, 181-190.
Gao, J. R., Zou, P. and Jiang, H. F. (1990). A new species of Luciaphorus
(Acariformes: Pygmephoridae) injuring Jew’s ear mushroom (Auricularia
polytricha) in China. Acta Agricultura Shangai 6, 68-7 1.
Gupta, P. C. and Chaudhry, H. S. (1972). New record of Hemitarsonemus latus
Banks (Tarsonemidae) as a parasite of Bemisia gossypiperda M. and L. Indian
Journal of Entomology 33, 476.
Hammen, L. van der (1980). “Glossary of Acarological Terminology”. Vol. 1.
“General Terminology. ” W. Junk, The Hague.
Herfs, A. ( 1926). Okologische Untersuchungen an Pediculoides ventricosus
(Newp.). Berl. Zoologica Stuttgart 74, 1-68.
Hirschfelder, H. and Sachs, H. (1952). Recent research on the acarine mite. Bee
World 33, 201-208.
Houck, M. A. and O’Connor, B. M. (1991). Ecological and evolutionary signifi-
cance of phoresy in the Astigmata. Annual Review of Entomology 36, 61 1-636.
Husband, R. W. (1984). Dipolipus, Panesthipolipus, Peripolipus and Stenopolipus,
new genera of Podapolipidae (Acarina) from the Indo-australian region.
International Journal of Acarology 10, 25 1-269.
Husband, R. W. and Sinha, R. N. (1970). A revision of the genus Locustacarus
with a key to genera of family Podapolipidae (Acarina). Annals of the Entomo-
logical Society of America 63, 1152-1 162.
Kaliszewski, M. (1984). Some aspects of the systematics and evolution of the
Tarsonemidae (Acari: Heterostigmata). In “Acarology VI” (D. A. Griffiths and
Bowman, C. E., eds), pp. 202-209. Ellis Horwood, Chichester.
Kaliszewski, M. and Wrensch, D. L. (1993). Evolution of sex determination and
sex ratio within the mite cohort Tarsonemina (Acari: Heterostigmata). In
“Evolution and Diversity of Sex Ratio in Insects and Mites” ( D. L. Wrensch
and M. E. Ebbert, eds), pp. 192-213. Chapman and Hall, New York.
Kantaratanakul, S. and Jitrat, S. (1984). Bionomics of the pyemotid mite Lucia-
phorus hauseri infesting cultivated jew’s ear fungus Auricularia polytricha in
Thailand. In “Acarology VI.” (D. A. Griffiths and C. E. Bowman, eds), pp.
622-628. Ellis Horwood. Chichester.
Kantaratanakul, S., Chaiwat, W. and Prayoonrat, P. (1989). Bionomics of the
dolichocybid mite, Formicomotes sp., as a pest of cultivated mushroom in
Thailand. In “Progress in Acarology” ( G. P. Channabasavanna and C. A.
Viraktamath, eds), pp. 223-228. Oxford University Press and IBH Publishing,
New Delhi.
Kosir, M. (1975). EmiihNng und Entwicklung von Pygmephorus mesembrinae und
P . quadratus (Pygmephoridae, Tarsonemini, Acari) und Bemerkungen iiber
weitere Arten. Pedobiologia 15, 3 13-329.
Krantz, G. W. (1957). Dolichocybe keiferi, a new genus and new species of
pyemotid mite, with a description of a new species of Siteroptes (Acarina:
Pyemotidae). Annals of the Entomological Society of America 50, 259-264.
Krantz, G. W. (1978). “A Manual of Acarology.” 2nd edn. Oregon State
University Book Stores, Corvallis.
Krczal, H. (1959)...Systematik und Okologie der Pyemotiden. In “Beitrag zur
Systematik und Okologie mitteleuroplscher Acarina,” Vol. 1, Abschn. 3 (H.
J. Stammer, ed.), pp. 385-625. Geest und Portig, Leipzig.
Le Fichoux, Y., Rack, G., Motte, P., Dellamonica, P. and Marty, P. (1980).
Dermatite prurigineuse due B Pyemotes zwoelferi Krczal, 1963. A propos de

plusieurs cas dans les Alpes-Maritimes. Acta Tropica 37, 83-89.
Lindquist, E. E. (1964). Mites parasitizing eggs of bark beetles of the genus Ips.
Canadian Entomologist 96, 125-126.
Lindquist, E. E. (1968). An unusual new species of Tarsonemus (Acarina:
Tarsonemidae) associated with the Indian honey bee. Canadian Entomologist
100, 1002-1006.
Lindquist, E. E. (1969a). New species of Tarsonemus (Acarina: Tarsonemidae)
associated with bark beetles. Canadian Entomologist 101, 1291-1314.
Lindquist, E. E. (1969b). Mites and the regulation of bark beetle populations. I n
“Proceedings of the 2nd International Congress of Acarology” (G. 0. Evans,
ed.), pp. 389-399. Akademiai Kiado, Budapest.
Lindquist, E. E. (1969~).Review of holarctic tarsonemid mites (Acarina: Prostig-
’
mata) parasitizing eggs of ipine bark beetles. Memoirs of the Entomological
Society of Canada no. 60, 1-1 11.
Lindquist, E. E. (1970a). Review of the genus Heterotarsonemus (Acarina:
Tarsonemidae). Canadian Entomologist 102, 8 12-829.
Lindquist, E. E. (1970b). Relationships between mites and insects in forest
habitats. Canadian Entomologist 102, 978-984.
Lindquist, E. E. (1973). Observations on the generic classification of tarsonemid
mites (Prostigmata). In: “Proceedings of the 3rd International Congress of
Acarology” (M. Daniel and M. and B. Rosicky, eds), pp. 293-295. W. Junk,
The Hague.
Lindquist, E. E. (1975). Associations between mites and other arthropods in forest
floor habitats. Canadian Entomologist 107, 425-437.
Lindquist, E. E. (1976). Transfer of the Tarsocheylidae to the Heterostigmata, and
reassignment of Tarsonemina and Heterostigmata to lower hierarchic status in
the Prostigmata (Acari). Canadian Entomologist 108, 2 3 4 3 .
Lindquist, E. E. (1983). Some thoughts on the potential for use of mites in
biological control, including a modified concept of “parasitoids.” In
“Biological Control of Pests by Mites” (M. A. Hoy, G.L. Cunningham and
L. Knutson, eds.), pp. 12-20. Division of Agricultural and Natural Resources,
University of California, Special Publication no. 3304.
Lindquist, E. E. (1985). Discovery of sporothecae in adult female Trochometridium
Cross, with notes on analogous structures in Siteroptes Amerling (Acari:
Heterostigmata). Experimental and Applied Acarology 1, 73-85.
Lindquist, E. E. (1986). The world genera of Tarsonemidae (Acari: Heterostig-
mata): a morphological, phylogenetic, and systematic revision, with a reclassi-
fication of family-group taxa in the Heterostigmata. Memoirs of the
Entomological Society of Canada no. 136, 1-517.
Lindquist, E. E. and Bedard, W. D. (1961). Biology and taxonomy of mites of the
genus Tarsonemoides (Acarina: Tarsonemidae) parasitizing eggs of bark beetles
of the genus Ips. Canadian Entomologist 93, 982-999.
Lindquist, E. E. and Kethley, J. B. (1975). The systematic position of the Hetero-
cheylidae Tragardh (Acari: Acariformes: Prostigmata). Canadian Entomologist
107, 887-898.
Lindquist, E. E. and Smiley, R. L. (1978). Acaronemus, a new genus proposed for
tarsonemid mites (Acari: Prostigmata) predaceous on tetranychoid mite eggs.
Canadian Entomologist 110, 655-662.
Lindquist, E. E., Kaliszewski, M. and Rack, G. (1990). Athyreacaridae, a new
family of mites (Acari: Heterostigmata) associated with scarab beetles of the

genus Neoathyreus (Coleoptera: Scarabaeidae). Acarologia 31, 161-176.
Magowski, W. (1988). Description of a new species of Formicomotes Sevastianov,
1980 (Acari, Dolichocybidae) with notes on the female dimorphism within the
genus. Mitteilungen aus dem Hamburger zoologischen Museum und Institut 85,
163-1 82.
Mahunka, S. (1970). Considerations on the systematics of the Tarsonemina and the
description of new European taxa (Acari: Trombidiformes). Acta Zoologica
Academiae Scientiarum Hungaricae 16, 137-174.
Mahunka, S. (1975). Neue und auf Insekten lebenden Milben aus Australien und
Neue-Guinea (Acari: Acarida, Tarsonemida). Annales Historico-Naturales
Musei Nationalis Hungarici 67, 3 17-325.
Mahsnka, S. (1981). Milben (Acari) aus St Lucia (Antillen). I. Tarsonemina,
Anoetidae. Acta Zoologica Academiae Scientiarum Hungaricae 27, 323-353.
Martin, N. A. (1978). Siteroptes (Siteroptoides) species with Pediculaster-like
phoretomorphs (Acari: Tarsonemida: Pygmephoridae) from New Zealand and
Polynesia. New Zealand Journal of Zoology 5 , 121-155.
Maynard Smith, J. (1989). “Evolutionary Genetics.” Oxford University Press,
Oxford.
Mitchell, R. (1967). Host exploitation of two closely related water mites. Evolution
21,59-75.
Morgenthaler, 0. (1934). Krankheitserregende und harmlose Arten der Bienen-
milbe Acarapis, zugleich ein Beitrage zum Species-Problem. Revue Suisse de
Zoologie 41, 4 2 9 4 6 .
Moser, J. C. (1975). Biosystematics of the straw itch mite with special reference to
nomenclature and dermatology. Transactions of the Royal Entomological
Society of London 127, 185-191.
Moser, J. C. (1981). Transfer of a Pyemotes egg parasite phoretic on western pine
bark beetles to the Southern pine beetle. International Journal of Acarology 7,
197-202.
Moser, J. C. and Cross, E. A. (1975). Phoretomorph: a new phoretic phase unique
to the Pyemotidae (Acarina: Tarsonemoidea). Annals of the Entomological
Society of America 68, 820-822.
Moser, J. C. and Roton, L. M. (1972). Reproductive compatibility between two
widely separated populations of Pyemotes scolyti (Acarina: Pyemotidae). Pan-
Pacific Entomology 48, 97-99.
Moser, J. C., Cross, E. A. and Roton, L. M. (1971). Biology of Pyemotes parvis-
colyti (Acarina: Pyemotidae). Entomophaga 16, 367-379.
Moser, J. C., Kielczewski, B., Wisniewski, J. and Balazy, S. (1978). Evaluating
Pyemotes dryas (Vitzthum 1923) (Acari: Pyemotidae) as a parasite of the
southern pine beetle. International Journal of Acarology 4, 67-70.
Moser, J. C., Smiley, R. L. and Otvos, I. S. (1987a). A new Pyemotes (Acari:
Pyemotidae) reared from the Douglas-fir cone moth. International Journal of
Moser, J. C., Smiley, R. L., Otvos, I, S. and Lorio, P. L. (1987b). Pyemotes n. sp.
(Acari: Pyemotidae), a potential biological control agent for cone-and-seed
insects. In “Proceedings of the 2nd IUFRO Conference on Cone and Seed
Insects Working Party” (A. Roques, ed.), pp. 307-308. BrianGon, France.
Newstead, R. and Duvall, H. (1918). Bionomic, morphological and economic
report on the acarids of stored grain and flour. Royal Society Reports on Grain
Pests (War) Committee no. 2, 1 4 8 .
Nickel, P. A. and Elzinga, R. J. (1969). Caraboacarus karenae, a new mite from

carabid beetle, with notes on its biology (Acari; Pyemotidae). Journal of Kansas
Entomological Society 42, 222-229.
Price, P. W. (1977). General concepts on the evolutionary biology of parasites.
Evolution 31, 405-420.
Rack, G. (1959). Acarophenax dermestidarum sp. n. (Acarina: Pyemotidae), ein
Eiparasit von Dermestes-Arten. Zeitschrift fur Parasitenkunde 19, 41 1 4 3 1.
Rack, G. (1966). Scutacaridae von Hamburg. 11. (Acarina, Trombidiformes).
Abhandlungen und Verhandlungen des Natumissenschaftlichen Vereins in
Hamburg 10,97-112.
Rack, G. (1967). Untersuchungen uber die Biologie von Dolichocybe Krantz 1957
und Beschreibung von zwei neuen Arten (Acarina, Pyemotidae). Mitteilungen
aus dem Hamburger zoologischen Museum und Institut 64, 29-42.
Rack, G. (1972). Pyemotiden an Gramineen in schwedischen landwirtschaftlichen
Betreiben. Ein Beitrag zur Entwicklung von Siteroptes graminum (Reuter, 1900).
(Acarina, Pyemotidae). Zoologisches Anzeiger 188, 157-174.
Rack, G. (1973). Dolichomotes crossi sp. n. aus Califomien und Bestimmung-
schliissel fur die Familie Dolichocybidae (Acarina, Pyemotoidea). Acarologia
15, 337-341.
Rack, G. (1975). Phoretisch auf Kleinsaugem gefundene Arten der Gattung
Pygmephorus (Acarina, Pygmephoridae). Mitteilungen aus dem Hamburger
zoologischen Museum und Institut 72, 157-176.
Rack, G. (1983). Beschreibung einer in Thailand an Speisepilzen (Auricularia
polytricha) schadlichen Milbe, Luciaphorus perniciosus sp. n. (Acarina: Tarso-
nemida, Pygmephoridae). Folia Entomologica Hungarica 44, 139-146.
Rack, G . and Eickwort, G. C. (1980). Biology and description of a new pygme-
phorid mite (Acarina: Tarsonemida) associated with the soil-nesting bee
Agapostemon nasutus (Hymenoptera: Halictidae). Acarologia 21, 267-278.
Rack, G. and Kaliszewski, M. (1985). Beschreibung des “normalen” und des
phoretomorphen Weibchens von Bakerdania aperta sp. n. (Acari; Pygmephor-
idae) aus Hamburg. Mitteilungen aus dem Hamburger zoologischen Museum und
Institut 82, 279-288.
Radowski, F. J. (1994). The evolution of parasitism and the distribution of some
dermanyssoid mites (Mesostigmata) on vertebrate hosts. In “Mites: Ecological
.and Evolutionary Analyses of Life History Patterns” (Houck, M. A., ed.) pp.
186-217. Chapman and Hall, New York.
Rakha, M. A. and Kandeel, M. H. (1983). Acarophenax meropsi n. sp., from the
European bee eater, Merops apiaster in Egypt, (Acari: Tarsonemida): Acarologia
24. 295-297.
Regenfuss, H. (1968). Untersuchungen zur Morphologie, Systematik und Okologie
der Podapolipidae (Acarina, Tarsonemini). Zeitschrift fur Wissenschafilische
Zoologie 177, 183-282.
Schousboe, C. ( 1 986). On the biology of Scutacarus acarorum Goeze (Acarina:
Trombidiformes). Acarologie 27, 15 1-158.
Smiley, R. L. and Landwehr, V. R. (1976). A new species of Tarsonemus
(Acarina: Tarsonemidae) predaceous on tetranychoid mite eggs. Annals of
the Entomological Society of America 69, 1065-1072.
Smiley, R. L. and Moser, J. C. (1976). Two new phoretomorphic Siteroptes from
galleries of the Southern pine beetle (Acarina: Pyemotidae). Beitruge zur
Entomologie, Berlin 26, 307-322.
Stannard, L. J. and Vaishampayan, S. M. (1971). Ovacarus clivinae (Acarina:
PARASITISM AND PARASITOIDISM IN TARSONEMINA 367
Podapolipidae), an endoparasite of the slender seedcom beetle. Annals of the

Steinkraus, D. C. and Cross, E. A. (1993). Description and life history of
Acarophenax mahunkai, n. sp. (Acari, Tarsonemina: Acarophenacidae), an egg
parasite of the lesser mealworm (Coleoptera: Tenebrionidae). Annals of the
Entomological Society of America 86: 239-249.
Suski. 2. W. (1973). A revision of Siteroptes cerealium (Kirchner) complex
(Acarina, Heterostigmata, Pyemotidae). Annales de Zoologie (Warsaw)-30,
509-535.
Tawfik, M. F. S. and Awadallah, K. T. (1971). The biology of Pyemotes herfsi
Oudemans and its efficiency in the control of the resting larvae of the pink
bollworms Pectinothra gossypiella Saunders, in UAR. Bulletin de la Sociktk
Entomologique dEgypte 54, 49-7 1.
Tawfik, M. F. S., El Husseini, M. M. and Awadallah, K. T. (1981). Interactions
between certain host larvae and the pyemotid ectoparasite, Pyemotes tritici.
Bulletin de la Sociitk Entomologique d’Egypte 63, 181-198.
Terry, J. R. (1966). “Studies of tarsonemid mites associated with bark beetles and
with rice.” MSc Thesis, La Trobe University.
Thorvilson, H. G. and Phillips, S. A., Jr (1987). The straw itch mite, Pyemotes
tritici (Acari: Pyemotidae), as a biological control agent of red imported fire
ants, Solenopsis invicta (Hymenoptera: Formicidae). Florida Entomologist 70,
439444.
Wehrle, L. P. and Welch, P. S. (1925). The occurrence of mites in the tracheal
system of certain Orthoptera. Annals of the Entomological Society of America
28,3544.
Wiggins, G. B., Mackay, R. J. and Smith, I. M. (1980). Evolutionary and ecological
strategies of animals in annual temporary pools. Archiv fur Hydrobiologie 58,
supplement, 97-206.
Zou, P., Gao, J. R. and Ma, E. P. (1993). Preliminary studies on the biology of the
pest mite Luciaphorus auriculariae (Acari: Pygmephoridae) infesting Jew’s ear
mushroom Auricularia polytricha in China. Experimental and Applied
Cumulative Index of Titles
Amino Acid Metabolism in Helminths, 30, 39

Anisakis and Anisakiasis, 16, 93
Anorexia: Occurrence, Pathophysiology, and Possible Causes in Parasitic
Infections, 24, 103
Argasid and Nuttalliellid Ticks as Parasites and Vectors, 24, 135
Arrested Development of Nematodes and some related Phenomena, 12, 179
Aspects of Acanthocephalan Reproduction, 19, 73
Aspects of the Host-Parasite Relationship of Plant-Parasitic Nematodes, 13, 225
Aspidogastrea, especially Multicotyle purvisi Dawes, 1941, 10, 78
Avian Blood Coccidians, 10, 1
Babesiosis: Non-specific Resistance, Immunological Factors and Pathogenesis, 17,

49
Bait Methods for Tsetse Fly Control, 34, 229
Behavioural Analysis of Nematode Movement, 13, 71
Biochemical Strain Variation in Parasitic Helminths, 25, 275
Biochemistry of the Variant Surface Glycoproteins of Salivarian Trypanosomes,
21, 69
Biological Aspects of Trypanosomiasis Research, 3, 1
Biological Aspects of Trypanosomiasis Research, 1965; a Retrospect, 1969,8,227
Biology and Distribution of the Rat Lungworm, Angiostrongylus cantonensis, and
its Relationship to Eosinophilic Meningoencephalitis and other Neurological
Disorders of Man and Animals, 3, 223
Biology of Fish Coccidia, 32, 293
Biology of the Acanthocephala, 5, 205; 11, 671
Biology of the Hydatid Organisms, 2, 169; 7, 327
Biology of Nanophyetus salmincola and “Salmon Poisoning” Disease, 8, 1
Biology of Pentastomids, 25, 45
Blastocystis in Humans and Animals: Morphology, Biology, and Epizootiology,
32, 1
Bmgian Filariasis: Epidemiological and Experimental Studies, 15, 243
Carbon Dioxide Utilization and the Regulation of Respiratory Metabolic Pathways

in Parasitic Helminths, 13, 35
Caryophyllidea (Cestoidea): Evolution and Classification, 219, 139
Cell-mediated Damage to Helminths, 23, 143
Cell-mediated Immunity Against Certain Parasitic Worms, 13, 183
370 CUMULATIVE INDEX OF TITLES
Cell-mediated Killing of Protozoa, 22, 43

Chagas Disease and Chagas Syndromes: The Pathology of American
Trypanosomiasis, 6, 63
Characterization of Species and Strains of Theileria, 26, 145
Chemical Communication in Helminths, 27, 169
Chemotherapy of Nematode Infections of Veterinary Importance, with Special
Reference to Drug Resistance, 35, 1
Circadian and other Rhythms of Parasites, 13, 123
Clonorchis and Clonorchiasis, 4, 53
Coccidia and Coccidiosis in the Domestic Fowl and Turkey, 1, 67
Coccidia and Coccidiosis in the Domestic Fowl, 6, 313
Conception and Terminology of Hosts in Parasitology, 14, 1
Control of Arthropods of Medical and Veterinary Importance, 11, 115
Copepoda (Crustacea) Parasitic on Fishes: Problems and Perspectives, 19, 1
Cryptobia and Cryptobiosis in Fishes, 26, 199
Cryptosporidiosis in Perspective, 27, 63
Cultivation of Helminths in Chick Embryos, 30, 107
Cultivation Procedures for Parasitic Helminths, 3, 159
Cultivation Procedures for Parasitic Helminths: Recent Advances, 9, 227
Current Concepts on the Biology, Evolution and Taxonomy of Tissue Cyst-forming
Eimeriid Coccidia, 20, 293
Dactylosomatidae, 30, 1
Distribution, Relationships and Identification of Enzymic Variants within the
Subgenus Trypanozoon, 29, 1
Dracunculus and Dracunculiasis, 9, 73
Dynamics of Parasitic Equilibrium in Cotton Rat Filariasis, 4, 255
East Cost Fever: Some Recent Research in East Africa, 15, 83

Echinococcus multilocularis Infection: Immunology and Immunodiagnosis, 31,
32 1
Echinostoma and Echinostomiasis, 29, 2 15
Ecological and Physiological Aspects of Helminth-Host Interactions in the
Mammalian Gastrointestinal Canal, 12, 183
Eggs of Monogeneans, 25, 175
Effects of Trypanosomatids on Insects, 31, 255
Electron Transport in Parasitic Helminths and Protozoa, 8, 139
Embryogenesis in Cestodes, 4, 107
Epidemiology of Amoebiasis, 6, 1
Epidemiology of Babesial Infections, 17, 115
Epidemiology and Control of Some Nematode Infections of Grazing Animals, 7,
211; 14, 355
Epidermis and Sense Organs of the Monogenea and Some Related Groups, 11, 193
Evasion of Immunity by Nematode Parasites Causing Chronic Infections, 26, 1
Evolutionary Biology of the Oxyurida (Nematoda): Biofacies of a Haplodiploid
Taxon, 28, 175
Evolutionary Trends in Mammalian Trypanosomes, 5, 47
Exoerythrocytic Development of Malarial Parasites, 27, 1
Experimental Chemotherapy of Schistosomiasis Mansoni, 6, 233; 12, 369
CUMULATIVE INDEX OF TITLES 371
Experimental Epidemiology of Hydatidosis and Cysticercosis, 15, 3 12
Experimental Fascioliasis in Australia, 7, 96
Experimental Research on Avian Malaria, 1, 1
Experimental Studies on Entamoeba with Reference to Speciation, 4, 1
Experimental Trichiniasis, 1, 213; 6, 361
Fascioliasis: the Invasive Stages of Fasciola hepatica in Mammalian Hosts, 2, 97

Fascioliasis: the Invasive Stages in Mammals, 8, 259
Feeding in Ectoparasitic Acari with Special Reference to Ticks, 3, 249
Fine Structure of the Monogenea especially Polystomoides Ward, 13, 1
Functional Morphology of Cestode Larvae, 11, 396
Giardia and Giardiasis, 17, 1

Giardia and Giardiasis, 32, 71
Genetic Basis of Diversity in Malaria Parasites, 22, 217
Genetic Control of Susceptibility and Resistance to Parasitic Infection, 16, 219
Genetic Diversity in Plasmodium falciparum, 29, 75
Global Problems of Imported Disease, 11, 75
Helminth Infections of Humans: Mathematical Models, Population Dynamics, and

Control, 24, 1
Hookworm Infection in Man, 17, 315
Host-Parasite Interface of Trematodes, 15, 201
Host-Parasite Relationships in the Alimentary Tract of Domestic Birds, 14, 96
Host-Parasite Relationships of Plant-Parasitic Nematodes, 7, 1
Host Specificity and the Evolution of Helminthic Parasites, 2, 1
Host Susceptibility to African Trypanosomiasis: Trypanotolerance, 21, 1
How Schistosomes Profit from the Stress Responses they Elicit in their Hosts, 35,
177
Hydatidosis/Cysticercosis: Immune Mechanisms and Immunization Against
Infection, 21, 229
Hydatidosis and Cysticercosis: The Dynamics of Transmission, 22, 26 1
Hypobiosis in Parasitic Nematodes - An Update, 25, 129
Immunology of Leishmaniasis, 32, 161

Immunity to Ticks, 18, 293
Immunity to Trypanosoma cruzi, 18, 247
Immunoelectron Microscopy of Parasites, 29, 151
Immunology of Schistosomiasis, 7, 41; 14, 399
Industrial Development and Field Use of the Canine Hookworm Vaccine, 16, 333
Infection Characteristics of Schistosoma japonicum in Mice and Relevance to the
Assessment of Schistosome Vaccines, 30, 167
Infectious Process, and its Relation to the Development of Early Parasitic Stages of
Nematodes, 6, 327
Infective Stage of Nematode Parasites and its Significance in Parasitism, 1, 109
Influence of Pollution on Parasites of Aquatic Animals, 30, 201
Interaction of Leishmania Species with Macrophages, 31, 175
Intestinal and Extraintestinal Life Cycles of Eimeriid Coccidia, 28, 1

Intramolluscan Inter-trematode Antagonism: a Review of Factors Influencing the
Host-Parasite System and its Possible Role in Biological Control, 10, 192
Larvae and Larval Development of Monogeneans, 1, 287; 6, 373

Leishmania, 2, 35
Lipid Metabolism in Parasitic Helminths, 22, 309
Liver Involvement in Acute Mammalian Malaria with Special Reference to
Plasmodium knowlesi Malaria, 6, 189
Lungworms of the Domestic Pig and Sheep, 11, 559
Malaria in Mammals Excluding Man, 5, 139 :

Metabolism of the Malaria Parasite and its Host, 10, 31
Metabolism of Entamoeba histolytica Schaudinn, 1903, 23, 105
Metacercarial Excystment of Trematodes, 33, 9 1
Meteorological Factors and Forecasts of Helminthic Disease, 7, 283
Minor Groups of Parasitic Platyhelminthes, 33, 145
Molecular Studies of Insect Vectors of Malaria, 34, 1
Molecular Mimicry, 34, 81
Myiasis of Humans and Domestic Animals, 35, 257
Nature and Action of Host Signals, 26, 239

Nature, Extent and Significance of Variation Within the Genus Echinococcus, 27,
209
Nematode Sense Organs, 14, 165
Nematodes as Biological Control Agents, Part I. Mermithidae, 24, 307
Nematodes as Biological Control Agents, Part 11, 31, 381
New Knowledge of Toxoplasma and Toxoplasmosis, 11, 631
Numerical Analysis of Enzyme Polymorphism; A New Approach to the
Epidemiology and Taxonomy of the Subgenus Trypanozoon, 18, 175
Onchocerciasis, 8, 173
Ontogeny of Cestodes and its Bearing on their Phylogeny and Systematics, 11,481
Oxygen-derived Free Radicals in the Pathogenesis of Parasitic Disease, 25, I
Paragonimus and Paragonimiasis, 3, 99; 7, 375

Paramphistomiasis of Domestic Ruminants, 9, 33
Parasite Behaviour: Understanding Platyhelminth Responses, 26, 73
Parasites and Complement, 27, 131
Parasites as Indicators of Water Quality and the Potential Use of Helminth
Transmission in Marine Pollution Studies, 35, 85
Parasitic Bronchitis, 1, 179; 6, 349
Parasitic Infections in Women and their Consequences, 31, 1
Parasitism and Commensalism in the Turbellaria, 9, 1
Pathogenesis of Mammalian Malaria, 10, 49
Parasitism and Parasitoidism in Tarsonemina (Acari: Heterostigmata) and
Evolutionary Considerations, 35, 335
CUMULATIVE INDEX OF TITLES 373
Pathophysiology of Malaria, 31, 843
Phylogeny of Life-cycle Patterns of the Digenea, 10, 153
Physiological Aspects of Reproduction in Nematodes, 14, 268
Physiological and Behavioural Interactions Between Parasites and Invertebrate
Hosts, 29, 271
Piroplasms: Life Cycle and Sexual Stages, 23, 37
Plasmodium Species Infecting Thamnomys rutilans: a Zoological Study, 33, 49
Post-embryonic Developmental Stages of Cestodes, 5, 247; 11, 707
Prevalence and Source of Toxoplasma Infection in the Environment, 28, 55
Problems in the Cultivation of some Parasitic Protozoa, 5 , 93
Prospects for the Development of Dead Vaccines against Helminths, 16, 165
Recent Advances in Antimalarial Chemotherapy and Drug Resistance, 12, 69

Recent Advances in the Anthelmintic Treatment of the Domestic Animals, 2, 221
Recent Experimental Research on Avian Malaria, 6, 293
Recent Observations on the Behaviour of Certain Trypanosomes within their Insect
Hosts, 22, 1
Recent Research on Malaria in Mammals Excluding Man, 11, 603
Recent Studies of the Biology of Trypanosoma v i v a , 22, 229
Regulation of Respiratory Metabolism in Parasitic Helminths, 16, 31 1
Regulatory Peptides in Helminth Parasites, 34, 163
Relationships between Chemotherapy and Immunity in Schistosomiasis, 34, 133
Relationship between Circulating Antibodies and Immunity to Helminthic
Infections, 8, 97
Relationships between the Species of Fasciola and their Molluscan Hosts, 3,59; 8,
25 1
Ribosomal RNA Genes of Plasmodium, 34, 33
Role of Tick Salivary Glands in Feeding and Disease Transmission, 18, 315
Sarcoptes scabiei and Scabies, 33, 235

Sarcosporidia (Protozoa, Sporozoa): Life Cycle and Fine Structure, 16, 43
Schistosoma mansoni: Cercaria to Schistosomule, 12, 115
Schistosomiasis and the Control of Molluscan Hosts of Human Schistosomes with
Particular Reference to Possible Self-Regulatory Mechanisms, 11, 307
Seasonal Occurrence of Helminths in Freshwater Fishes, Part I, Monogenea, 15,
133; Part 11, Trematoda, 17, 141; Part 111, Larval Cestoda and Nematoda, 18, 1;
Part IV, Adult Cestoda, Nematoda and Acanthocephala, 20, 1
Serotonin Receptors in Parasitic Worms, 23, 2
Sexual Development of Malarial Parasites, 22, 153
Sexuality of Parasitic Crustaceans, 32, 367
Snail Control in Trematode Diseases: the Possible Value of Sciomyzid Larvae
Snail-Killing Diptera, 2, 259
Snail Problems in African Schistosomiasis, 8, 43
Some Tissue Reactions to the Nematode Parasites of Animals, 4, 321
Speciation in Parasitic Nematodes, 9, 185
Species of Leucocytozoon, 12, 1
Structure and Composition of the Helminth Cuticle, 4, 187
Structure of the Helminth Cuticle, 10, 347
Taeniasis and Cysticercosis (Taenia saginata), 10, 269

Taxonomy and Transmission of Leishmania, 16, 1
Tick Feeding and Its Implications, 8, 275
Toxoplasma and Toxoplasmosis, 5 , I
Transport of Nutrients and Ions across Membranes of Trypanosomatid Parasites,
32, 261
Transmission of Parasites Across the Placenta, 21, 155
Treatment of Human African Trypanosomiasis, 33, 1
Trichomonas vaginalis and Trichomoniasis, 6, 1 17
Trichostrongyloid Nematodes and Their Vertebrate Hosts: Reconstruction of the
Phylogeny of a Parasitic Group, 24, 239
Trichuris and Trichuriasis in Humans, 28, 107
Trypanosomes of Anura, 11, 1
Uitrastructure of the Tegument of Schistosoma, 11, 233
Vaccination Against the Canine Hookwork Disease, 9, 153

Variation in Echinococcus: Towards a Taxonomic Revision of the Genus, 35, 145
Vector Relationships in the Trypanosomatidae, 15, 1
Veterinary Anthelmintic Medication, 7, 350
Cumulative Index of Authors
Adamson, M.L., 28, 175 Cambell, W.C., 35, 1

Adler, S., 2, 35 Cameron, T.W.M., 2, 1
Aikawa, M., 29, 151 Chabaud, A., 33,49
Alexander, J., 31, 175 Chubb, J.C., 15, 133; 17, 141; 18, 1;
Alicata, J.E., 3, 223 20, 1
Anderson, R.M., 24, 1 Clark, G.W., 10, 1
Anya, A.O., 14, 267 Clarke, I.A., 25, 1
Arthur, D.R., 3, 249; 8, 275 Clegg, J.A., 16, 165
Athias-Binche, F., 35, 335 Coleman, G.S., 18, 121
Atkinson, C.T., 29, 151 Conder, G.A., 35, 1
Constantine, C.C., 35, 145
Cooper, E.S., 28, 107
Cowden, W.R., 25, 1
Baker, J.R., 10, 1 Cowman, A.F., 29, 75
Baker, R.D., 29, 1 Croll, N.A., 13, 71
Ball, S.J., 28, 1; 32, 293 Crampton, J.M., 34,1
Bardsley, J.E., 11, 1 Crompton, D.W.T., 14, 95; 19, 73
Barrett, J., 30, 39
Barta, J.R., 30, 1
Beesley, W.N., 11, 115 Davies, A.J., 32, 293
Behnke, J.M., 26, 1 Dawes, B., 2, 97; 8, 259
Bennett, G.F., 10, 1 De Jong-Brink, M., 35, 177
Berg, C.O., 2, 259 Denham, D.A., 15, 244
Beme, A.D., 8, 43 Desser, S.S., 12, 1
Bertram, D.S., 4, 255 De Vasconcellos Coelho, M., 16, 1
Binnington, K.C., 18, 315 DOMelly, J., 17, 115
Bishop, A,, 5, 93 Durette-Desset, M.-C., 24, 239
Blackwell, J.M., 22, 43
Boray, J.C., 7, 95 Ellis, D.S., 22, 1
Boreham, P.F.L., 32, 1 Elsdon-Dew, R., 6, 1
Brabin, B.J., 31, 1 Erasmus, D.A., 15, 201
Brabin, L., 31, 1 Evans, D.A., 22, 1
Brener, Z., 18, 247
Brindley, P.J., 34,133
Brocklesby, D.W., 17.49 Fallis, A.M., 12, 1
Bruce-Chwatt, L.J., 11, 75 Fletcher, A., 10, 31, 49
Bryant, C., 8, 139; 13, 36; 16, 31 1; 25, Flockhart, H.A., 25,275
275 Frayha, G.J., 22, 309
Bundy, D.A.P., 28, 107 Freeman, R.S., 11, 481
Burgess, I., 33, 235 Fried, B., 27, 169; 29, 215; 30, 107; 33,
Butterworth, A.E., 23, 143 91
376 CUMULATIVE INDEX OF AUTHORS
Garcia, E.G., 30, 167 Khan, R.A., 12, 1; 30, 201

Gardiner, P.R., 28, 229 Knapp, S.E., 8, 1
Garnham, P.C.C., 5, 139; 11, 603 Koberle, F., 6, 63
Gemmel, M.A., 15, 312; 22;259 Komiya, Y., 4, 53
Gibbs, H.C., 25, 129
Gibson, W.C., 18, 175 Laarman, J.J., 20, 293
Gibson, T.E., 2, 221, 7, 350 Laird, M., 10, 1
Godfrey, D.G., 18, 175; 29, 1 Landau, I., 33, 49
Gottstein, B., 31, 321 Larsh, J.E., Jr., 1, 213; 6, 361; 13, 183
Green, C.H., 34, 229 Lawson, J.R., 22, 261
Lee, D.L., 4, 187; 10, 347
Hall, M., 35, 257 Leid, R.W., 27, 131
Hall, R., 34, 81 Liew, F.Y., 32, 161
Halton, D.W., 34, 163 Lim, H.K., 10, 192
. Hansen, E.L., 9, 227 Lindquist, E.E., 35, 335
Harmsen, R., 11, 1 Llewellyn, J., 1, 287; 6, 373
Haseeb, M.A., 27, 169 Loke, Y. W., 21, 155
Hawking, F., 13, 123 Long, P. L., 1, 67; 6, 313; 28, 1
Heydorn, A.O., 16.43 Lumsden, W. H. R., 3, 1; 8, 227
Heyneman, D., 10, 191 Lymbery, A.J., 27, 209; 35, 145
Ho, M., 31, 83 Lyons, K.M., 11, 193
Hoare, C.A., 5, 47
Hockley, D.J., 11, 233 McGreevy, P.A., 15, 243
Hominick, W.M., 31, 381 McLaren, D.J., 14, 195
Hoogstrall, H., 24, 135 MacKenzie, K., 35, 85
Horak, I.G., 9, 33 McVicar, A.H., 35, 85
Horton-Smith, C., 1, 67; 6, 313 Mackiewicz, J.S., 19, 139
Huff, C.G., 1, 1; 6, 293 Maegraith, B., 6, 189; 10, 31, 49
Huffman, J.E., 29, 215 Mansour, T.E., 23, 1
Hughes, D.L., 2, 97; 8, 259 Marshall, T.F. DE C., 18, 175
Hutchison, W.M., 28, 55 Maule, A.G., 34, 163
Hunt, N.H., 25, 1 May, R.M., 24, 1
Hurd, H., 29, 271 Mehlhorn, H., 16, 43; 23, 37
Mehlitz, D., 29, 1
Inglis, W.G., 9, 185 Meis, J.F.G.M., 27, 1
InGn, A.D., 26, 145 Mendis, A.H.W., 32, 71
Mettrick, D.F., 12, 183; 26, 73
Jackson, M.H., 28, 55 Meyer, E.A., 17, 1
Jacobs, L., 5, 1; 11, 631 Michel, J.F., 7, 211; 12, 280; 14, 355
Jerinings, J.B., 9, 1 Millemann, R.E., 8, 1
Jirovec, O., 6, 117 Miller, T.A., 9, 153; 16, 333; 17, 315
Johnstone, P.D., 15, 312 Milord, F., 33, 1
Joyner, L.P., 17, 115 Mitchell, G.F., 30, 167
Molyneux, D.H., 15, 1
Kabata, Z., 19, 1 Morrison, W.I., 21, 1
Kaliszewski, M., 35, 335 Muller, R., 9, 73
Katz, N., 6, 233, 12, 369 Murray, M., 21, 1
Keam, G.C., 25, 175
Kemp, D.H., 18, 315 Neal, R.A., 4, 1
Kemp, D.J., 29, 75 Nelson, G.S., 8, 173
Kendall, S.B., 3, 59; 8, 251 Nesheim, M.C., 14, 96
CUMULATIVE INDEX OF AUTHORS 377
Nicholas, W.L., 5, 205; 11, 671 Smithers, S.R., 7, 41; 14, 399
Smyth, J.D., 2, 169; 7, 327; 22, 309
Odening, K., 14, 1 Sommerville, R.I., 1, 109; 6, 327; 26,
O’Donnell, C.A., 32, 161 239
Ollerenshaw, C.B., 7, 283 Stableford, L.T., 30, 107
Stenzel, D.J., 32, 1
Parshad, V.R., 19, 73 Stirewalt, M.A., 12, 115
Pawlowski, Z., 10, 269 Sukhdeo, M.V.K., 26, 73
Pearson, J.C., 10, 153 Symons, L. E. A., 24, 103
Pellegrino, J. 6, 233; 12, 369
Pepin, J., 33, 1 Tadros, W., 20, 293
Peters, W., 12, 69 Terry, R.J., 7, 41; 14, 399
Petersen, J.J., 24, 307 Thomas, J. D., 11, 307
Pet& M., 6, 117 Thompson, R. C. A., 27,209; 32.71; 35,
Pittilo, R.M., 28, 1 145
Podesta, R.B., 12, 183 Thome, K. J. I., 22, 43
Popiel, I., 31, 381 Thulin, J., 30,201
Poynter, D., 1, 179; 4, 321; 6, 349 Tiu, W. U., 30, 167
Pumell, R.E., 15, 83 Trilles, J. P., 32, 367
Tumer, M. J., 21, 69
Radulescu, S., 17, 1 Tzipori, S., 27, 63
Raibaut, A., 32, 367
Reeves, R.E., 23, 105 Verhave, J. P., 27, 1
Reynoldson, J.A., 32, 71 Voge, M., 5, 247; 11, 707
Rickard, M.D., 21, 229
Rickman, L.R., 29, 1
Riley, J., 25, 45 Wakelin, D., 16, 219
Rogers, W.P., 1, 109; 6, 327; 26, 239 Wall, R., 35, 257
Rohde, K., 10, 78; 13, 1; 33, 145 Walliker, D., 22, 217; 29, 75
Rose, J.H., 11, 559 Waters, A.P., 34,33
Russell, D.G., 31, 175 Weatherly, N.F., 13, 183
Rybicka, K., 4, 107 Webster, J.M., 7, 1; 13, 225
White, N.J., 31, 83
Schaub, G. A., 31,255 Willadsen, P., 18, 293
Schein, E., 23, 37 Williams, B., 35, 85
Schultz, M. G., 10, 269 Williams, H.H., 35, 85
Shaw, C., 34, 163 Williams, J.F., 21, 229
Siddall, R., 35, 85 Williams, P., 16, 1
Silverman, P. H., 3, 159; 9, 227 Whitelaw, D.D., 21, 1
Sinclair, I.J., 8, 97 Woo, P.T.K., 26, 199
Sinden, R.E., 22, 153 Wootten, R., 16, 93
Slais, J., 11, 395
Smart, D., 34, 163 Yokogawa, M., 3,99; 7, 375
Smith, J.W., 16,93
Smith, L.P., 7, 283 Zilberstein, D., 32, 261
Smith, M.A., 16, 165 Zwart, D., 17,49
Index
Abamectin 14-15 Arenivaga apacha 346

Abothrium gadi 91 Artemia salina 7
Abramis brama 95. Ascaridia galli 25
Acanthocephala, life cycles 125 Ascaris suum 38
Acanthocephalus anguillae 96 Ascogaster reticulatus 229
Acarapis 359 Asiocortarsonemus 356
Acarapis externus 350, 355 Aspidogastrea, life cycles 121
Acarapis woodi 356, 360 Auchmeromyia 266, 269
Acaronemus 355 Auchmeromyia senegalensis 268
Acarophenax 352
Acarophenar meropsi 353 Bacillus thuringiensis 57, 299
Acarophenar nidicolus 350 Bacillus thuringiensis var wuhhanensis
Acarophenax tribolii 353 299
Actinotrichida 336 Bacterial clearance assay (BCA) 193
Adactylidium 352, 353 Bakerpolipus 357
Adactylidium mahunkai 353 Barbara colfaiana 35 1
Adenylate cyclase (AC) 207, 213, 214 Bemisia tabaci 354
Adrenocorticotropic hormone (ACTH) Benzimidazoles
220,225,226, 232 mechanism of resistance 46-8
Aeromonas salmonicida 193 resistance to 27
Agapostemon nasutus 339 Biomphalaria 202
Alanine-proline-glycine-tryptophan Biomphalaria glabrata 189, 191, 192,
(APGW) neurons 205, 214 195,197,201,202,204,225,226
Albendazole 5 Biomphalaria pfeizeri 202
Alphitobius diaperinus 353 Blennius pholis 90
Amcortarsonemus 356 Blood parasites in fish 89-90
y-aminobutyric acid (GABA) 5 Boophilus microplus 22
Ancylostoma caninum 14, 19 Bos indicus 287
Androgens 221 Bothriocephalus acheilognathi 92, 95
Anguilla rostrata 99 Bothriocephalus scorpii 91
Anthelmintics Buccinum undatum 94, 126
currently approved compounds 3-4 Bucephaloides gracilescens 119
delivery methods 4 Bunostomum phlebotomum 20
resistance to 27
Anti-oxidant enzymes 184 Cachexin 233
Apanteles kariyai 230 Caenorhabditis elegans 5 , 6, 25, 48
Apatemon gracilis 98 Calfluxin (CaFl) 204,205,206,210,
Apis cerana 355 212,218,222
Apis mellifera 356 Calliphora 260
Archipolipus 357 Calliphora albifrontalis 286
380 INDEX
Calliphora nociva 286 Cochliomyia minima 280

Calliphora stygia 283, 286 Cololabis saira 121
Calliphora vicina 282 Cooperia curticei 24
Calliphora vomitoria 282, 285-6 Cooperia oncophora 20, 2 1, 34
Calliphoridae Cooperia pectinata 20
current status of species 279-86 Cooperia punctata 20
evolution 271-3 Cooperia spatulata 20
insecticide control 296-8 Cooperia spp. 11, 35
pathology and immunology 291-2 Cooperia surnabada 20
predisposing conditions for myiasis Cordylobia 266
288-9 Cordylobia anthropophaga 268, 286,
principal species and life cycles 292, 298
266-9 Cordylobia myiasis 286
Canopy cells (CCs) 208, 214, 215 Cordyfobia rodhaini 269, 286
Caryophyllaeus laticeps 96, 97 Coreitarsonemus anoplocnemis 356
Catostomus commersoni 93 Corticotropin releasing factor (CRF) 220
Caudodorsal cell hormone (CDCH) 204, Cotesia congregata 230
207, 209, 210, 224 Cotesia glomerata 23 1
Caudodorsal cells (CDCs) 204,212,213, Cotylurus sp. 98
222-3, 224 Cryptolestes 353
Cephalopina titillator 263, 290, 294 Cucullanus heterochrous 9 1
Cephenemyia 262 Cuterebra 262, 266, 272
Cephenemyia auribarbis 263 Cuterebra angustifrons 290
Cephenemyia trompe 263, 276, 310 Cuterebra beameri 290
Cercaria-derived factor 2 18, 2 19 Cuterebra buccata 29 1
Cercaria doricha 119, 126 Cuterebra latifrons 27 1
Cercaria pythionike 1 19 Cuterebrinae 2654,277-9
Cercariae 2 18 Cyprinus carpio 93, 101
Chaetogaster limnaei limnaei 98 Cytokines 220, 233
Chilled Fly Dispersal System (CFDS)
30 1 Dactylogyrus 96
Chorioptes bovis 11 Dactylogyrus falcatus 96
Chrysomelobia 357 Dactylogyrus wunderi 96
Chrysomya albiceps 28 1 Damalinia bovis 11, 21
Chrysomya bezziana 262, 266-7, 272, Dendroctonus frontalis 352
280-1, 288, 291, 296, 301, 307, Depolarizing afterpotential (DAP) 2 12
308, 310 Dermutobia 272
Chrysomya megacephala 273, 28 1 Dermatobia hominis 2 1, 265, 270, 27 1,
Chrysomya putoria 28 1 277-9, 287, 290-2, 295-6, 298,
Chrpomya rufifacies 28 1-2, 283 299, 300
Clean water stimulus (CWS) 223 Dermatophilus congolensis 288
CLIMEX in forecasting 308 Detoxifying enzymes 184
Clonostachydiol 26 Diclidophora merlangi 88, 89, 111
Clonostachys cylindrospora 26 Dictyocaulus viviparus 11, 20, 21, 34-5
Clupea harengus 120 Dictyocotyle coeliaca 112
Coccipolipus 357 Diffusible molecules 187-90
Cochliomyia hominivorar 2 1, 262, Digenea, life cycles 122
266-7, 269, 272, 279-80, 287, Dilopolipus 357
288, 291, 296, 299, 300-1, 305, Dioxapyrrolomycin 25-6
307, 308, 310 Diplostomum sp. 96, 98
Cochliomyia macellaria 273, 280, 282 Dirofilaria immitis 14, 19
INDEX 38 1
Dolichocybe hippocastani 343, 344 Species 1 162-4
Dolichocybe keiferi 343 Species 2 164
Dolichocyboidea 343 Species 3 165
Dolichomotes crossi 343 Species 4 165
Doramectin 20-2 Species 5 166
formulation and pharmacokinetics 20 Species 6 166
in cattle 20-2 Species 7 166
Dorsal bodies (DBs) 204 species concepts and their application
Dorsal body hormone (DBH) 204, 207, 150-2
210 species definition 150-1
Dorsipes 357 species status 152
Dotting immunoassay (DIA) 218, 223 taxonomic considerations 147-50
Drug delivery, new methods 4-5 taxonomic revision 145-76
Drug resistance variation in 146-7
and management practices 45 Echinococcus equinus 165
antinematodal drugs 26-57 Echinococcus granulosus 146, 147,
causes of treatment failure unrelated 153-8, 164
to 3 8 4 0 buffalo strain 154, 166
continuing spread of 42-3 camel strain 155-6, 162
definition 26 cattle strain 155, 164
extent worldwide 29-38 cervid strain 156-7
factors contributing to 43-5 horse strain 154-5, 165
factors in assessment of 40-1 lion strain 157
growth in 28 miscellaneous strains 157
identification of 40 pig strain 156, 162
in cattle 34-5 sheep strain 153, 166
in goats 29-34 Tasmanian sheep strain 154, 166
in horses 35-8 Echinococcus multilocularis 146, 158-9,
in sheep 29-34 165-6
in swine 38 Echinococcus multilocularis
mechanisms of 45-9 kazakhensis 158
monitoring/reporting 49-53 Echinococcus multilocularis
multigeneric 28 multilocularis 158
strategies to limit development 54-7 Echinococcus multilocularis sibiricemis
Drugs, for use against nematode 158
parasites in various animal Echinococcus oligarthrus 146, 159, 166
species Echinococcus ortleppi 164
3 4 Echinococcus vogeli 146, 159, 166
Echinoparyphium recurvatum 94, 98
Ecdysteroids 188 Echinorhynchus gadi 91, 92
Echeneibothrium 92 Echinostoma liei 95, 229
Echinobothrium 92 Echinostoma paraensei 192
Echinococcus Echinostoma revolutum 98
current classification of species and Eicosanoids 189
strains 148-9 Eimeria adenoeides 232
delimitation of evolutionary species Electrophysiological studies 212-1 3
162-7 P-endorphin 225
delimiting species 152 Entobdella soleae 110
evolutionary species concept 151-2 Enzyme-cytochemical studies 2 13-14
identification of OTUs 152-9 Epistylis sp. 95
phylogeny of OTUs 159-62 Ergasilus sieboldi 96
382 INDEX
Eucestoda, life cycles 117 Glutathione transferases (GSTs) 184

Eustrongylides sp. 95 Glyphodomastax 346
Eutarsopolipus 357 Gonadotropin-releasing hormone
(GnRH) 221
Fasciola hepatica 9 Grillotia erinaceus 112, 1 15
Female-killing (FK) system 302 Grillotia smaris-gora 115, 121
Fish Growth hormone (GH) 221
blood parasites in 89-90 Growth hormone releasing hormone
gill parasites in 88-9, 100 (GHRH) 221
gut parasites in 9&2 Gut parasites, in fish 90-2
heavy metal pollution in 92-4 Gyrocotylidea, life cycles 114
miscellaneous pollution effects Gyrodactylus macrochiri 95
98-105 Gyrodactylus spp. 88
skin parasites in 88-9, 100 Gyrodactylus stellatus 95
.thermal pollution in 9 4 8 Gyrostigma 269
Fish parasites
guidelines and selection procedures
126-8 Habronema muscae 19
hydrocarbon pollution effects on Haematobia irritans 11
87-92 Haematopinus eurysternus 21
pollution effects via invertebrate Haematopinus suis 13
intermediate hosts and non- Haemogregarina bigemina 90
piscine Haemonchus contortus 6, 21, 23-6, 34,
vertebrate hosts 102 38,42,45-9,51-3, 55
Fluorescein isothiocyanate (FITC) Haemonchus placei 20
210-1 1 Haemonchus similis 21
Follicle-stimulating hormone (FSH) 22 1 Haemonchus spp. 34
Formicomotes heteromorphus 343 Heat shock proteins (HSPs) 185-7
Fundulus heteroclitus 95 Heavy metal pollution, in fish 92-4
Heliothis virescens 230
GABA (y-aminobutyric acid) 5 Helminth life cycles, and transmission
Gadus morhua 88 processes 109-26
Gammarus pulex 99 Helminth transmission, in marine
Gasterophilinae 263, 2 7 4 5 pollution studies 85-144
Gasterophilus 262, 294, 3 10 Hemisarcoptes cooremani 340
Gasterophilus haemorrhoidalis 263, Heterakis spumosa 25
264 Heterocheylus sp. 344
Gasterophilus inermis 264 Heterotarsonemus 339
Gasterophilus intestinalis 19, 264, 275, Histioplasma 339
‘306 Hormone-like secretory substances
Gasterophilus nasalis 19, 264 183-90
Gasterophilus nigrocornis 264 Hormone-receptor complexes 2 10-12
Gasterophilus pecorum 263, 264, 27 1 HSP 70 237
Gastrophilus intestinalis 13 Human chorionic gonadotropin (hCG)
Gastrophilus nasalis 13 232
Gedoelstia 263 Human growth hormone (hGH) 228
Genetically impaired female technique Hydatid disease
(GIFT) 302 control of 146-7
Gill parasites 88-9, 100 status of 146
Glugea stephani 95 Hydrocarbon pollution effects on fish
Glutathione peroxidase (GPx) 184 parasites 87-92
INDEX 383
Hymenolepis diminuta 230 Locustacarus 357
Hypoderma 270, 27 1,275-7 Luciaphorus auriculariae 348
Hypoderma bovis 21,264-5,276, 294, Luciaphorus hauseri 348
300 Luciaphorus perniciosus 348
Hypoderma diana 265 Lucilia 310
Hypoderma lineatum 264-5, 290, 294, Lucilia bufonivora 273
299,300, 393 Lucilia caesar 282-3
Hypoderma spp. 44,310 Lucilia cuprina 232, 262, 267, 272,
Hypoderma tarandi 276-7, 306 283-5,288-9,292,296,298,
Hypodermatinae 264-5, 275-7 299, 301,303,304,305,308
Hypothalamic-pituitary-gonad (HPG) Lucilia illustris 282
axis 219,221 Lucilia sericata 262, 267, 272, 282-5,
Hysterothylacium aduncum 9 1 288-9,308-9, 31 1
Luteinizing hormone (LH) 221, 232
Ichthyocotylurus erraticus 98 Lymnaea stagnalis 99, 180, 193,
Ichthyocotylurus sp. 96 199-203,205. 206,209, 214-18
Ichthyophthirius multifiliis 93, 101 effects of Trichobilharzia ocellata
Ictalurus punctatus 93 190-204
11-6 232
Immune system (IS) 231-5 Macrocheles muscaedomesticae 299
suppressive effect of stress on 220-1 Macrocyclic lactones
Insecticides 293-8 mechanism of action 5-7
Internal defence system (IDS) 222-6, structures of 8
23 1-5 Marine parasites
Iponemus 341, 352, 354, 358, 359 effects of natural environmental
Ivermectin 6
factors 105-9
antiparasitic utility 9 principal biotic and abiotic factors
formulations 9
general properties 7-8 106
Marine pollution studies, helminth
in cattle 9-11, 10 transmission in 85-144
in dogs 13-14 Megaselia 260, 261
in horses 13, 14 Meriones unguiculatus 23
in sheep 11, 12 Merlangius merlangus 88
in swine 11-13 Metabolic enzymes 184
Microplitis croceipes 229
Johanssonia arctica 88 Micropterus salmoides 94
Juvenile hormones (JHs) 189 Milbemycin B-41D 19
Milbemycin D 6
Laciaphorus perniciosus 347 Milbemycin oxime in dogs 19
Lacistorhynchus sp. 121 Molluscan insulin-related peptides
Lacistorhynchus tenuis 112-14 (MIPS) 208
Lasiusflavus 339 Monogenea, life-cycles 113
Lecithochirium furcolabiatum 120 Moxidectin 6
Lepocreadium setiferoides 123-5 antiparasitic utility 15
Lepomis macrochirus 95, 101 general properties 15
Leuckartiara 98 in cattle 15-18
Levamisole, mechanisms of resistance in horses 18-19
48 in sheep 18
Libyostrongylus douglassi 38 in swine 18
Light green cells (LGCs) 212-15, 223 wMSH 226
Linognuthus vituli 2 1 M l T 193
384 INDEX
Mucor miehei 7 Oesophagostomum columbianum 24

Musca autumnalis 17 Oesophagostomum dentatum 38
Musca domestica 260, 299 Oesophagostomum quadrispinulatum 38
Myiasis 257-34 Oesophagostomum radiatum 20
classification 259-62 Oestridae
control techniques 293-305 current status of species 273-9
baits, traps and targets 305 evolution 269-71
biological 299 insecticide control 293-6
genetic 301-3 pathology and immunology 289-91
insecticides 293-8 predisposing conditions for myiasis
mechanical 298 287-8
sterile insect technique (SIT) principal species and life cycles
299-30 1, 303 262-6
vaccines 3 0 3 4 sterile insect control technique
I definition 258-9 299-30 1
evolution 269-73 Oestrinae
forecasting 308-9 current status of species 2 7 3 4
immunology 289-92 principal species and life cycles
modelling 308-9 262-3
monitoring Oestrogens 22 1
by serodiagnosis 306 Oestrus 262, 310
by traps 307-8 Oestrus ovis 1 1,262-3,2734,289,293,
pathology 289-92 294, 299, 306
physiology 287-92 Oligocottus maculosus 89
predisposing conditions for 287-9 Onchocerca cervicalis 13
principal species and life cycles Onthophagus gazella 15
262-9 Ostertagia circumcincta 2 1, 24, 5 1, 55
taxonomic groupings 26 1 Ostertagia lyrata 20
Myoxocephalus octodecemspinosus 89 Ostertagia ostertagi 20, 2 1, 25, 34
Myoxocephalus scorpius 9 1 Ostertagia spp. 1 1, 34
Mytilus edulis 99 Oxyuris equi 19
Mytilus galloprovincialis 99
Mytilus trossulus 99
Panesthipolipus 357
Paracarophenax 353
Nemadectin 15 Paradiplozoon homoion 97
Nematoda, life cycles 124 Paraherquamide
Nematode infections efficacy in vitro and in laboratory
chemotherapy 1-84 animals 23
drug resistance 1-84 general properties 22
drbgs currently approved 3 4 in cattle 24
Nematodirus helvetianus 20-1 in dogs 24
Nematodirus spathiger 20, 50 in sheep 2 3 4
Neoechinorhynchus cylindratus 94 safety and other aspects 24-5
NES 222-6, 2314, 235 structure 22
effects of parasite-derived factors Parapygmephorus costaricanus 339
228-3 1 Parascaris equorum 19
Neuroendocrine system (NES) 180 Parasites
Neuropeptide Y (NPY) 220-1 as long-term stressors 219-34
Nomia melanderi 345 factors interfering with production
Notocotylus attenuatus 93, 94 and action of host regulatory
Notocotylus urbanensis 98 substances 229-3 1
INDEX 385
factors mimicking host regulatory Psorergates ovis 11, 17

substances 228 Psoroptes bovis 2 1
see also Fish parasites; Marine Psoroptes ovis 11
parasites PTTH-ecdysone-JH system 23 1
Parasitism 335-67 Pyemotes barbara 35 1
Parasitoidism 335-67 Pyemotes dryas 352
Parophyrys vetulus 95 Pyemotes herfsi 352
Pectinophora gossypiella 35 1 Pyemotes parviscolyti 349, 352
Pediculaster jlechtmanni 338 Pyemotes scolyti 349
Penicillium charlesii 22 Pyemotes tritici 35 1, 352
Penicillium paraherquei 22 Pyemotes ventricosus 349
Pennella sp. 121 Pyemotoidea 349-53
Peripqlipus 357 Pygmephoridae 336
Peromyscus leucopus leucopus 290 Pygmephoroidea 348
Perperipes 346 Pygmephorus mesembrinae 348
Petalomiumjimbrisetum 339 Pygmephorus punctatus 347
PF1022A 25
Phagocytosis assay (PA) 193 Raja clavata 92
Pharyngomia picta 263 Regenopolipus 357
Phoresy 337-41 Restriction fragment length
Phormia regina 285, 31 1 polymorphism (RFLP) 153
Physa gyrina 97 Rhinoestrus 294
Physogastry 336, 341 Rhinoestrus latifrons 274
Pimephales promelas 92 Rhinoestrus purpureus 263, 274
Piophila 260 Rhipidocotyle campanula 101
Pityophthorus bisulcatus 349 Rhipidocotyle sp. 118
Planorbarius corneus 195-7, 2 16 Rhynchopolipus 357
Planorbis corneus 226 Ribosomal DNA (rDNA) 153
Plasmodiumfalciparum 185
Pleuronectes platessa 89, 118 Sarcocystis cruzi 233
Podapolipidae 357 Sarcophaga 260
Podapolipoides 357 Sarcophaga cruentata 268
Podapolipus 357 Sarcophagidae 288-9, 291-2
Podocotyle atomon 91 current status of species 286-7
Polydnaviruses (PDV) 230 evolution 27 1-3
Polymerase chain reaction (PCR) 218 insecticide control 296-8
Polyphagotarsonemus latus 354 principal species and life cycles
Pomphorynchus laevis 99 266-9
Popilius disjunctus 355 Sarcopromusca 266
Pro-opiomelanocortin (POMC) 220, Sarcopromusca arcuata 287
225, 226 Sarcoptes scabiei 11, 21
Prostaglandins (PGs) 189 Schistosoma 178
Proteocephalus neglectus 121 Schistosoma mansoni 186, 189, 191,
Proteolytic enzymes 184 192, 195-9,201, 202,204,
Protocalliphora 268, 269 2254,234
Protophormia terraenovae 282-3, 285 Schistosome-snail partnerships
Przhevalskiana silenus 277, 295 effects on internal defence 190-200
Pseudocarapis 355 effects on metabolism and growth
Pseudoletia separata 230 2024
Pseudomonas aeruginosa 288 effects on reproduction 201-2
Pseudopleuronectes americanus 90 Schistosomes 177-256
386 INDEX
acquisition of energy and space Streptomyces avermitilis 20

179-80 Streptomyces cyaneogriseus
as stressors for vertebrate hosts 227-8 noncyanogenus 15
effects on regulatory systems of hosts Streptomyces hygroscopicus
190 aureolacrimosus 19
excretory products 183 Stress
immunoevasion strategies 179 and immune system 220-1
interference with host’s regulatory and reproduction and growth 221
systems 180-2 concept in mammals 219-21
parasitic components/products as physiological 23 1
candidates for interference with Stress proteins 185
regulatory mechanisms in host Strongyloides papillosus 20
182-90 Strongylus edentatus 19
problems to be solved by 178-80 Stroigylus vulgaris 13, 19
. secretory products 183 Superoxide dismutases (SODS) 184
tegument and surface coat
components 182 Tarsocheyloidea 336
Schistosomiasis 178 Tarsonemella 355
Schistosomin 207 Tarsonemina 335-67
induction of release by parasite- distribution of physogastry, parasitism
derived factor 216-19 and parasitoidism among
inhibiting effect on CDCs 213 341-57
interaction with hormone-receptor Tarsonemoidea 354-7
complexes 210-12 Tarsonemoides sp. 350
origin of 214-16 Tarsonemus 339
primary structure 208 Tarsonemus dispar 355
purification and structure 209-10 Tarsonemus sp. 350
Scolytus multistriatus 349 Tarsopolipus 357
Scolytus ventralis 349 Tenebrio molitor 230-1
Scomber scombrus 115 Testosterone 232
Screw-worm adult suppression system Tetrapolipus 357
(SWASS) 305 Thelazia spp. 21
Scutacaroidea 346 Thermal pollution, in fish 94-8
Scutacarus acarorum 339 Throchometridium tribulatum 345
Scutacarus palustris 347 TNF 233
Siteroptes cerealium 348 TNFa 232
Siteroptidae 348 Tolypocladium niveum 299
Skin parasites in fish 88-9, 100 Toxascaris leonina 14
Solenopotes capillatus 17, 21 Toxocara canis 14, 25
Solenopsis invicta 35 1 Toxocara cati 25
Somatomedin C 233 Trachurus trachurus 115
Sphaerostoma bramae 96 Trematodes 178
Spirometra mansonoides 228 Tribolium 353
Sprattus sprattus 126 Trichobilharzia cameroni 98
Squatina squatina 116 Trichobilharzia ocellata 180, 188, 189,
Stenopolipus 357 197-200, 203
Stephanostomum baccatum 118, 126 as stressor for Lymnaea stagnalis
Sterile insect technique (SIT) 302 222-6
Steringophorus furciger 90, 91 effect on reproduction and growth of
Stichocotyle nephropis 118 snail host 204-19
Stigmurcarus 357 effects on Lymnaea stagnalis 190-204
INDEX 387
effects on reproduction and growth of Trypanosoma murmanensis 90
snail host, central effects 212-14 Turritella 98
mode of interference with growth and Tylodelphys clavata 96
metabolism 207-8
mode of interference with Uncinaria stenocephala 14, 19
reproduction 204-7
peripheral effects on snail host Vasoactive intestinal peptide (VIP) 229
204-12 Vulpes vulpes 282
Trichobilharzia ocellata-Lymnaea
stagnalis 190 Water quality, parasites as indicators of
Trichostrongylus aexi 20, 24, 34 85-144
Trichostrongylus colubriformis 6, 20, Wohlfahrtia magnifica 262, 267, 272,
2 3 4 4 6 - 9 , 53 286-7,289, 291,297, 305
Trichostrongylus longispicularis 20 Wohljahrtia meigeni 268, 273
Trichostrongylus spp. 34 Wohlfahrtia nuba 267
Trichotromidium muscarum 299 Wohlfahrtia vigil 268
Trichuris discolor 2 1
Trichuris spp. 11, 20 Xenopus laevis 5
Trwhometridioidea 345-6
Trochometridium sp. 344 Zoarces viviparus 91
Trypanosoma brucei 232 Zoogonus lasius 125

(Advances in Parasitology 31) J.R. Baker and R. Muller (Eds.) - Academic Press (1992)

Загружено:

Сведения о документе

Оригинальное название

Авторское право

Доступные форматы

Поделиться этим документом

Поделиться или встроить документ

Параметры публикации

Этот документ был вам полезен?

Это неприемлемый материал?

Авторское право:

Доступные форматы

(Advances in Parasitology 31) J.R. Baker and R. Muller (Eds.) - Academic Press (1992)

Загружено:

Авторское право:

Доступные форматы

Editorial Board

C. Bryant Department of Zoology, Australian National University, G.P.O.

C. Combes Laboratoire de Biologie Animale, UniversitC de Perpignan,

M. Coluzzi Director, Instituto di Parassitologia, Universita Delgi Studi di

W.H.R. Lumsden 17A Merchiston Crescent, Edinburgh EHlO 5AX, UK

E.J.L. Soulsby Department of Clinical Veterinary Medicine, University of

K.Tanabe Laboratory of Biology, Osaka Institute of Technology, Ohmiya,

K.S. Warren Comprehensive Medical Systems, Inc., 46 1 Fifth Avenue,

P. Wenk Tropenmedizinisches Institut, Universitat Tubingen, D7400

United States Edition published by

This book is printed on acid-free paper

All Rights Reserved

Typeset by J&L Composition Ltd, Filey, North Yorkshire

Genetics and Animal Disease, School of Veterinary Studies, Murdoch

Clearly, if parasites are to be useful as indicators of water quality a sound

catastrophic effects if it reached countries south of the Sahara. Fortunately,

The Charles A . Dana Research Institute for Scientists Emeriti. Drew

ADVANCES IN PARASITOLOOY VOL 35 Copyright Q 1995 Academic Press Limited

The present time seems especially appropriate for a review of antinematode

circumstances. As discussed below, known or anticipated drug resistance

3. NEW METHODS OF DRUG DELIVERY

consisting of three layers, with morantel tartrate incorporated in the middle

4.1. Mechanism of Action

Haemonchus contortus. A similar pattern of ivermectin sensitivity in

Figure 1 Time-dependent effects of 0.1 p~ ivennectin (0).avermectin B1 (m),

macrocyclic lactones against nematodes was due to inhibition of chitin

Ivermectin is the 22, 23-dihydro derivative of the natural product aver-

4.2.1. General Properties

Figure 2 Structures of macrocyclic lactones.

Ivermectin is absorbed when given orally or parenterally, the rate of

4.2.2. Antiparasitic Utility

and inhibited (hypobiotic) stages. An unexpected feature of the efficacy

" Oral formulation.

intended for incorporation into swine feed, so as to give a dosage of

2. HEARTGARD-30 Chewable: disks of soft moist material containing

Abamectin or avermectin B, is the naturally occurring precursor of

Moxidectin is a derivative of nemadectin, a naturally occurring product of

4.4.1. General Properties

4.4.2. Antiparasitic Utility

1. CYDECTIN Injection: a sterile solution containing 1% moxidectin. It is

injectable preparation) resulted in 100% elimination of the mites and

" oral formulation.

findings of the study were considered preliminary because various batches,

4.5. Milbemycin B-41D

The milbemycins are produced by Streptomyces hygroscopicus aureola-

4.6. Milbemycin Oxime

Dogs. Milbemycin oxime (Figure 2) is an 80:20 mixture of the & and

dosage used commercially. The following product is approved for

helvetianus. Trials against infections in cattle in Latin America confirmed

Paraherquamide (Figure 3) is an oxindole alkaloid of fungal origin. It was

Figure 3 Structures of paraherquamide, PF1022A, dioxapyrrolomycin, and

5.1.1. Efficacy in vitro and in Laboratory Animals

5.1.2. Efficacy in Sheep

5.1.3. Efficacy in Cattle

5.1.4. Efficacy in Dogs

5.1.5. Safety and Other Aspects

The above data suggest paraherquamide could be used successfully

PF1022A (Figure 3) is a cyclodepsipeptide of fungal origin. It was isolated

Dioxapyrrolomycin (Figure 3) was originally isolated as a metabolite from

Clonostachydiol (Figure 3) was ,identified as a metabolite of the fungus,

6. RESISTANCE TO ANTINEMATODAL DRUGS

6.1. Is Resistance an Issue?

Drug resistance is here defined as: a heritable reduction in the sensitivity of