SOVE 1993, VOL 18, NO 2.pdf

BSOVE 18( 2): 75- 187 ( 1993)
ISSN 0146- 6429
Bulletin of the
SOCIETY FOR
VECTOR ECOLOGY
4" 4
JIL
Volume 18, Number 2
December, 1993 f
BULLETIN OF THE
SOCIETY FOR VECTOR ECOLOGY
Volume 18- Number 2 December 1993
Marc J. Klowden, Editor

Division of Entomology
University of Idaho
Moscow, ID 83844-2339
Phone: ( 208) 885- 7546
Fax: ( 208) 885- 7760
EDITORIAL BOARD
Mir S. Mulla, Chairman
University
of
California
W. J. Crans
R. S. Nasci
Rutgers
CDC-DVBID
University
Riverside, CA 92521
New Brunswick, NJ
R. C. Axtell
C. I. Dahl
North Carolina State
University
University
Fort Collins, CO
M. W. Service
of
Liverpool School of Tropical Med.
Uppsala
Raleigh, NC
Uppsala, Sweden
Liverpool, England
N. Becker
F. R. Holbrook
E. D. Walker
USDA- ARS
Laramie, WY
Michigan State University

East Lansing, MI
D. R. Barnard
R. S. Lane
S. K. Wikel
Med. Vet. Entomol. Lab
University of California
Berkeley, CA
Oklahoma State University
USDA, Gainesville, FL
H. Briegel
G. G. Marten
S. C. Williams
New Orleans MCD
San Francisco State University
CH- 8057, Zurich, Switzerland
New Orleans, LA
San Francisco, CA
E. P. Catts
L. S. Mian
University
Germany
University
of
of
Heidelberg
Zurich
Washington State
Stillwater, OK
County Vector Control
University
Pullman, WA
San Bernardino, CA
Published by the Society for Vector Ecology

to disseminate pertinent information from all
facets of the field of vector ecology and related disciplines.

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should be addressed to the Editor. Communications relating to galley proofs,
reprints, subscriptions, SOVE membership, change of address, and other matters
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PUBLICATIONS AND BUSINESS OFFICE
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BOARD OF DIRECTORS
1993 OFFICERS
President
Norbert Becker
President- Elect
Susan M. Palchick
Vice- President
Bruce F. Eldridge
Past- President
William K. Reisen
Secretary- Treasurer
Gilbert L. Challet
REGIONAL DIRECTORS
Southwestern
Minoo B. Madon
Northwestern
Steven V. Romney
North Central
Thomas R. Wilmot
South Central
Cluff E. Hopla
Northeastern
Wayne J. Crans
Southeastern
Jonathan F. Day
European
Frantisek Rettich
PUBLICATIONS AND BUSINESS OFFICE
Society for Vector Ecology

P. O. Box 87
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Membership Plus Bulletin$ 45.00

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Sustaining Members$ 150. 00
BULLETIN
OF THE

Number 2
December, 1993
Volume 18
CONTENTS
ii
Guidelines for Contributors

In Memoriam- Thomas D. Mulhern ( 1908In Memoriam- John M. Doll( 1935-
ix
1993)
1993)
xi
In Memoriam- Tuti R. Hadi( 1943 - 1993)
xii
Book Review
Arthropods of Medical and Veterinary Importance: A Checklist of Preferred Names and Allied Terms. A.
R.
Reviewed
Pittaway
by
D. L. Baumgartner
75
Proceedings
7th SOVE Annual European Branch Meeting, Bologna, ITALY, 25 August 1992
Tick Ecology and the Search for a Possible Lyme Disease Spirochaete in Southeastern Australia
Richard C. Russell, S. L. Doggett,
and
R. Munro
77
First International Congress of Vector Ecology, San Diego, California, 4 October 1993
The Role
of
Ecology
and
Ecological
Modeling
M. W. Service
in Vector Control
85
Submitted Papers
Borrelia burgdorferi Infection in Ixodes scapularis ( Acari: Ixodidae) in Kent County Maryland
F. P. Amerasinghe
and
T. W. Scott
99
Size of Emerging and Host Seeking Aedes aegypti and the Relationship to Containers and Blood-Feeding
D. D. Chadee
Success in Trinidad, West Indies
105
Daily Survivorship and Life Span of the Mosquito Eretmapodites quinquevittatus Theobald ( Diptera:
Culicidae) Under Laboratory Conditions Compared toAedes aegypti,Aedes abopictus, andAedes bahamensis
D. Vodopich
109
J. D. Lang
County, California
Elevated Dosages of Bacillus thuringiensis var. israelensis Fail to Extend Control of Culex Larvae
M. S. Mulla, J. D. Chaney, and J. Rodcharoen
114
A. M. Helleck, W. K.
Fleas Found
on
Blow
and
Mammals in San Diego
The Oriental Latrine
Invading
Hartberg,
Fly
of
Fly,
Chrysomya
megacephala (
Public Health Importance
Fabricius 1794) ( Diptera: Calliphoridae),
A. R. Olsen, T. H. Sidebottom,
and
125
as an
S. G. Bennett
133
Preliminary Research on Bacillus thuringiensis var. israelensis Use for Mosquito Control in Northeast
K. Ouzounis
Greece
and
A. Samanidou- Voyadjoglou
147
Mosquito Utilization of Resting Sites at an Urban Residence in Southern California

E. T. Schreiber, W. E. Walton,
and
M. S. Mulla
152
Field Trials of a Fizzy Tablet With Bacillus thuringiensis subsp. israelensis in Forest Spring Ponds in
A. G. Eriksen
160
Occurrence and Abundance of Cyanobacteria in a Brackish Marshland and Their Ingestibility by Mosquito
I. Thiery, G. Sinegre, and N. Tandeau De Marsac
Larvae
164
O. Skovmand
Denmark
and
Water Depth and Larval Density Affect Development and Accumulation of Reserves in Laboratory
Populations
of
Mosquitoes
S. E. Timmennann
and
H. Briegel
174
BULLETIN OF THE SOCIETY FOR VECTOR ECOLOGY
Guidelines for Contributors
The Bulletin of the Societyfor Vector Ecology is an international journal concerned with
all aspects of the biology, ecology, and control of arthropod vectors and the interrelationships
between the vectors and the disease agents they transmit. Thejournal publishes original research
articles and research notes, as well as comprehensive reviews of vector biology based on
presentations at Society meetings. All papers will be reviewed by at least two referees who are
qualified
scientists and who recommend their
suitability for
publication.
Acceptance of
manuscripts is based on scientific merit and is the final decision of the editor, but these decisions
may be appealed to the Editorial Board.
Scientific contributions should be sent to Dr. Marc J. Klowden, Editor, Division of

Entomology, University of Idaho, Moscow, Idaho 83843, U.S. A. Manuscripts must be double
spaced on a single side of bond paper with 25 mm margins. An original and two clear copies
are required. Draft mode dot matrix type should not be used. Submission of text on a 3- 1/ 2"
computer
diskette formatted in MS- DOS is
encouraged.
Microsoft Word, Word Perfect, or
Wordstar formats are acceptable, as well as unformatted text files. Please indicate the type of
format on the diskette label. Papers must be organized under the following headings, each on
a separate page, in order: Title page, abstract, text, acknowledgments( ifappropriate), references
cited, tables, figure legends, and figures. The title page should contain the names of all authors
and their affiliations and the
identification
and address of the
corresponding
author.
Pages
should be numbered consecutively starting with the title page. References should conform to
the style in recent volumes. Illustrations that are submitted must be clearly labeled and legible
after reduction.
Page charges, which partially defray the cost ofpublication, are$ 35 per printed page.
Reprint charges are shown in the table below.
Pages
50
copies
1- 4
5- 8
60. 00
20.00
9- 12
13- 16
17- 20
105. 00
150. 00
195. 00
240. 00
30.00
40. 00
50.00
60.00
or less
Each
add'
50 copies
Same order
ii
NUMBER 2
DECEMBER, 1993
VOLUME 18
IN MEMORIAM
Thomas Desmond Mulhern

1908 - 1993
rh
4 nRnx
He was a driving force within the Eastern
If ever the letter" M," as in mosquito, were applicable

to one whose name began
with
was
Thomas D. Mulhern,
devoted
control.
and
careers, one which
Association of Mosquito Control Workers, predecessor
his lifetime
to the American Mosquito Control Association, which
mosquitoes and mosquito
originated in New Jersey. Tommy served the American
M,"
Mulhern had two

began in New
originated, and the other
such was
who throughout
dedicated to
Tommy
the case
with"
mosquito control
Mosquito Control Association as its President in 1968
he
and 1969 and as its Executive Director during the later
Jersey,
from
which
years of his career.
in California.
performed technical
In 1949, Tommy was invited to California to
New Jersey Agricultural Experiment
While in New Jersey,
Tommy
control
consider a position with the state' s newly organized

Bureau of Vector Control. He arrived for a look-see in
mosquito
mid-July at the peak of daily temperatures and mosquito
utilizing the New Jersey light

trap. This subsequently led to development of amodified
unit, the American light trap.
production and was escorted throughout the state to
services within the
Station,
which
commissions.
benefited the local
He
assisted
in the
mosquito
state' s
surveillance activities,
familiarize him
with
the problems.
This failed to
dismay him as he shortly returned with wife Helen and

ix
NUMBER 2
family
with
DECEMBER, 1993
household
goods to
stay. He
astonishment over the comparative size of

and
California, in
that
New
Jersey
could
admitted to
he was an enthusiastic participant in various aspects of
New
Jersey
the state, national, and world vector control programs.
within the
Tommy was the recipient of the first Honorary

Membership offered by the Society for Vector Ecology
fit
boundary of California' s largest county.

Tommy and family purchased a ranch- style home
in 1977.
in Fresno, from which location he carried out his activities

statewide until
his
retirement as
Specialist in 1975.
In
order
Tommy was invited by the World Health
Senior Vector Control

to
utilize
Organization to perform a number of consultations to

other nations for the benefit of their vector control
his broad
background in mosquito control, Tommy was designated
programs
Technical Consultant in Mosquito Control,

serving
within the Bureau' s Headquarters' staff. He performed
various
technical
services
mosquito control agencies,
VOLUME 18
for the benefit
of
Sadly, Helen, his devoted and supportive lifetime
companion, predeceased Tommy a year ago. Tommy
local
passed
largely relating to mosquito
away
on
12 August 1993, he
was
85.
He is
survived by their three sons, Thomas, John, and Martin.
control and source reduction equipment. He assisted in
the development of the training program for local vector

control
technicians
to fulfill requirements of the
Richard F. Peters
Environmental Protection
Agency. While required to

work at least 40 hours
weekly, Tommy characteristically
Retired Chief
nearly doubled that expectation. Throughout his
California State Department of Public Health
Bureau of Vector Control
career
VOLUME 18
DECEMBER, 1993
NUMBER 2
IN MEMORIAM
John M. Doll
1935 - 1993,,
40
4441
off
4111100,
We
passed
John
are sad to announce that
away
From 1974 to 1993, John was the supervisor of the
at the age of 58.
Vector- Borne and Zoonotic Diseases Section of the
up in Colorado and earned a bachelor of

degree in entomology from Colorado State
University
He
Sunday,
Dr. John M. Doll
November 21,
grew
science
army
on
in 1957.
He
went
on
to earn
in the
Fort Detrick, Maryland.
borne disease surveillance and control in Arizona. He
then served four years
as a research scientist at
Arizona Department of Health Services( ADHS). John
built the vector program at ADHS from the ground up

and devoted much energy toward improving vector-
a masters
degree in
medical
was highly respected and popular among his colleagues
entomology from the University of Delaware in 1962.

From 1962 to 1967, John worked for the New Mexico
and friends both in and outside of public health.
State Health Department
son- in- law Mike, and six grandchildren. John will be
In 1968, John
and
his
He is survived by his wife Shirley, daughter Karol,
as a vector control specialist.
family
moved to
Syracuse, New
sorely missed.
York, where he worked as senior medical entomologist

for the New York State Health Department. In 1970,
Craig E. Levy
John
Vector-Borne and Zoonotic Diseases Section
went
back to
school and earned
microbiology in 1974,
at the
his doctorate in
University
of
Arizona.
Arizona Department of Health Services
xi
DECEMBER, 1993
VOLUME 18
NUMBER 2
IN MEMORIAM
Tuti R. Hadi
1943 - 1993
Ihor
Indonesia
irreplaceable
Hadi
and
the
asset on
jaw
of the
rest
world
13 March 1993,
when
Tuti Hadi
lost
an
mammalogistsanddevelopedcollaborativestudieswith
Dr. Tuti
some of them. From 1976 to 1978, she worked as a
born
Senior Technician and then as a Research Scientist at
although
theU.S. Naval Medical Research Unit No. 2( NAMRU-
con-
2) in Jakarta, an institution with which she continued
the
to keep close ties. At NAMRU-2, she collaborated on
Indonesian Archipelago and was also a visiting scientist
epidemiological and parasitological surveys through-
on
succumbed to meningitis.
25 July 1943, in
the rest of her
ducted her
life
research
Bandung,
centered
Java,
around
throughout the
in the United States. Most
of
degrees,
was completed
a second masters
Hawaii
at
American
Java,
she
wide span of
including
and doctoral(
out Indonesia with Pat Camey, John Cross, David
1989)
Dennis, Dirk van Peenan, and others. From 1976 until
in Jakarta,
the time of her death, she continued with similar studies
However, she also completed
as a Research Scientist and then as Senior Research
in Javanese
Magelang, orYogyakarta.
and
her training,
bachelor' s( 1966), master' s( 1968),
was
degree ( 1980)
schools
University
of
Scientist at the National Institute for Health Research
many
and Development, Ministry of Health, also in Jakarta.
medical entomologists, parasitologists, and
Dr. Hadi' s research was published in more than 40 peer-
Manoa. Tuti Hadi
at the
was well- known to
Xii
DECEMBER, 1993
NUMBER 2
reviewed articles.
expertise
is
The breadth
reflected
of
in the topics
her interests
of some of
Lance A. Durden
and
Institute of Arthropodology and Parasitology

Georgia Southern University
these
ranging from island surveys for vector- borne

diseases ( notably, scrub typhus, murine typhus, and
works
Landrum Box 8056
Statesboro, GA 30460
plague) and their reservoir hosts, mammals, and their

parasites (
especially
ectoparasites),
VOLUME 18
the effect of
Indonesian transmigration policies on disease

epidemiology, and systematics of parasitic mites.
Perhaps her
most
detailed
works pertained
typhus ( which remains a scourge
Southeast Asia)
vectors.
and to the
With the passing
nesian scientific
in many
taxonomy
of
of
Guy G. Musser
Department of Mammalogy,
American Museum of Natural History,
to scrub
parts
of
the chigger
Central Park West at 79th St.,
Tuti Hadi, the Indo-
community has lost its
New York, NY 10024
major source
of expertise in acarology and one of its leading vector

biologists. Indonesia and the rest of the world has lost
a dedicated scientist and it is hoped that other researchers
in Indonesia and elsewhere will be
some
of her important
We
are grateful to
Pat W. Carney
able to continue with
Schistosomiasis Research Project,
work.
Mohammad Amir, Head
Vector Biology and Control,
of the
1901 Fort Myer Drive, Suite 402,
Museum Zoologicum Bogoriense in Java for supplying

biographical information
on
Arlington, VA 22209
Dr. Hadi.
xiii
DECEMBER, 1993
BULL. SOC. VECTOR ECOL., 18( 2): 75- 76
BOOK REVIEW
Arthropods of Medical and
A. R.
and
important animals.
Importance: A
Veterinary
Checklist of Preferred Names
The book is divided into sections entitled arthropods,
Allied Terms'
microorganisms,
Pittaway
arthropod- transmitted
viruses,
Publisher: CAB International, United Kingdom; U. S.
entomogenous fungi, helminths, larvivorous fish, and
1991
other organisms( such as planarians), all with a consistent
University of Arizona Press, Tucson,
Distributor:
format of the scientific name of the organism arranged
178 pp.,$ 28. 50

ISBN: 0- 85198- 741- 9
alphabetically in the left margin, followed by the author

of that species name.
To the right is the family and
order/class name to which each species belongs and,
While perusing medical/ veterinary researchreports

all
where applicable, the correct and currently recognized
professionals have encountered unfamiliar species names
name if the one to the left is an incorrect and antiquated
may have been spelled slightly differently in two

different publications and/ or noticed that the author of
synonym. In total, about 7, 500 entries are included.

All the scientific names provided were derived
the species name was also different. Some editors and
from CAB Abstracts, an abstracting database maintained
meticulous about
by CAB International, which is the publisher of the
and
literature,
other
time or another
one
at
which
their selected
peer reviewers are
very
spelling of scientific names included in their

journals, but other editor/ reviewers are quite lax,
Review of Medical and Veterinary Entomology
the proper
especially those who may review papers mentioning

the journal' s specialty. I have
species that are outside
found that this is

journals
parasitology,
papers
often the case
textbooks
and
dealing
nonentomological
veterinary,
occasionally may include
which
etc.)
for
( medical,
with arthropods of medical/ veterinary
formerly the Review of Applied Entomology Series

B).
Since 1913, CAB' s staff have maintained and
regularly updated a list of all scientific names that have

been recorded within its Abstracts. These names are
consistently checked against internationally recognized

references or by leading specialists. This meticulous
attention to detail is quite evident in this book.
it is
The commendable attention to detail is evidenced
time consuming to locate and
handy, and relatively inexpensive reference book
also by the surprisingly few typographical errors or

inconsistencies detected. Approximately 500 scientific
names, including many of the most commonly
encountered medically/ veterinary important arthropods,
were checked for accuracy by comparing the listed
these
names to those provided in other popular medical
The Arthropods of Medical and

Importance:
A Checklist ofPreferredNames
Veterinary
Insects and Related Organisms( 1989, Entomological
importance.
When these discrepancies
difficult
sometimes
and
search other works that

scientific name
A remedy
single,
has been
in
may
order to
cite the particular species'
verify its accuracy.
finally
to these problems
written
that
arise,
is
specifically
One
available.
addresses
problems with names.
and
Allied Terms
source
was compiled
as a reliable
Society
of
America).
The only discrepancies were
scientific names and taxonomic
found in six species in which the presentation of authors'
the majority of important species and genera
names were different from that cited in the ESA
for verifying the
position of
to serve
entomology textbooks and to the Common Names of
of arthropods worldwide
in the fields
veterinary entomology.
Its intent is to introduce
of medical and
publication ( usually the inclusion or exclusion of

parentheses around
the author).
In all cases, the latter
measure of consistency to these names, allowing accurate
publication was really in error when other reliable
be
sources were consulted for a third opinion. However, a
of
few medically important arthropods were absent from

Pittaway' s book which should have been included: the
and reliable
information regarding
Also included
communicated.
biological
control agents(
employed
to
manage
intermediate hosts,
including
these
certain
diseases in
selected
lists
fish) that have been
arthropod
natural
organismscarriedbythesemedical/
that may cause
nomenclature to
are
vectors,
enemies,
and
veterinary arthropods
man
and
economically
saddleback caterpillar(
Io
moth ( Automeris
garden spider(
Sibine
stimulea[
Clemeus]), the
io [ Fabricius]), the white-backed
Argiope
trifasciata [ Forskal]),
tick( Haemaphysalis chordeilis[
the bird
Packard]), the clematis
Reviewed by D. L.Baumgartner, William Rainey Harper College, Department of Biological Sciences, 1200 West
Algonquin Road, Palatine, Illinois 60067, U. S. A.
76
BULL. SOC. VECTOR ECOL.
blister beetle( Epicauta
Lytta
vesicatoria [
cinerea [ Forster]),
Linn.]),
and
Spanish
fly
any Neotropical
Compsomyiops Townsend blowflies. Putting

omissions aside, the coverage of species
is
these few
otherwise
commendable.
Judging
No work is without flaws, however, and this book

is no exception. The title is inappropriate and somewhat
misleading.
From previewing the title only, many
prospective buyers may presume that the contents

consist of common names of species, when in fact
from its
contents, the
primary
audience of
this book would be nonentomological researchers and
doctors,
practitioners, such as medical
veterinarians,
parasitologists, public health professionals, pest control
technicals, and wildlife rehabilitators who

otherwise
DECEMBER, 1993
have
limited knowledge
would
not
single
common
name
is listed.
Additional
criticisms include the small size of type chosen for the
book, even when only a few lines of text are printed on

a single page, such as on the inside title page and in the
table of contents, preface, and acknowledgments. The
but
decision to utilize a small print for the species names is
may commonly or occasionally encounter them in

their line of work.
Inadvertently, professional articles
understandable and justified in order to conserve the
of arthropods
who
frequently
read
by
these professionals may
list various arthropod speciesnames,

earlier
inaccuracies. To these
would
be
an
orsimply propagate
professionals, this work
invaluable reference.
total number of printed pages, and thereby the cost, but

people with vision impairments may experience
difficulty in reading the book as a result.
In summary, this book is of limited usefulness to
Similarly, the listing
the professional and experienced medical/veterinary
unaccepted species synonyms
entomologist. However, putting aside the few problems
prove useful to entomologists who read or consult
noted, this inexpensive book will serve as an
of archaic and
may
incorrectly
older
literature
taxon.
limited
currently
and are not specialists
Otherwise,
this reference
usefulness to
entomologists.
The
practicing
book
in that particular
would
medical and
be
who possess minimum training in entomology, such as
veterinary
medical doctors, veterinarians, parasitologists, public

health officials, pest control technicians, and wildlife
listing of families and orders

belong is of little usefulness
which these arthropods
to
to
the competent entomologist, but again, it

may be useful
to other professionals who
arthropods.
indispensable reference for researchers and practitioners
of only
occasionally" dabble"
with
rehabilitators, as well as a variety of other individuals

who may have a need to refer to a definitive book on the
correct scientific names of the major arthropods of
importance.
medical/ veterinary
DECEMBER, 1993
TICK ECOLOGY AND THE SEARCH FOR A POSSIBLE LYME DISEASE

SPIROCHAETE IN SOUTHEASTERN AUSTRALIA'
R. C. Russell2, S. L. Doggett2, and R. Munro3
ABSTRACT: Lyme disease has been reported from Australia based on clinical and serological diagnoses, but a
causative organism has not been identified. Attempts were made to detect, isolate, and identify spirochaetes from
ticks collected in areas associated with putative human infections. Ticks were collected by dragging in natural
bushland and by removal from native and domestic animals. They were dissected and a portion of the gut was
examined for spirochaetes by microscopy, with the remainder inoculated into culture media. The cultures were
examined for spirochaetes by microscopy and with immunochemical and molecular( PCR) techniques. Whole ticks
were also tested with PCR for spirochaetes. Between 1990 and 1992, more than 20,000 ticks were collected and
approximately 12, 000 were processed for spirochaetes. No evidence of Borrelia burgdorferi or other spirochaete
was detected. Some spirochaete-like objects detected in culture were shown to be artifacts, probably aggregates of
bacterial flagellae. The ecology of Ixodes holocyclus, the tick most likely to be involved as a vector to humans is
discussed in the context of a possible local spirochaetosis resembling Lyme disease.
INTRODUCTION
unpublished
data).
It is well recognized that there are
problems of specificity and sensitivity associated with

Lyme disease has been
from North
and also
serological testing for Lyme disease( Sigal and Curran

1991); this is particularly true for Australia where no
South- East Asia, South America, Africa, and Australia
local causative spirochaete has been isolated for use as
reported
America, Europe, Russia, China,

However, there
Sigal 1988).
doubt
as
to the
hemisphere in
southern
The first
particular.
in Australia
reported
Japan
remains considerable
Lyme disease in the
true
existence of
and
in Australia in
and
general,
Between January 1990 and December 1992, a field

and laboratory program was undertaken to identify
disease'
vectors of the putative disease through isolation of the
diagnoses from the
causative agent B. burgdorferi or similar spirochaete.
cases of putative ` Lyme

were clinical
a reference antigen.
South Wales
This would provide unequivocal evidence for the local
N. S. W.) in southeastern Australia( Stewart et al. 1982).
existence of an etiological agent for the disease. Local
later reported from the south
spirochaetes could then be used as antigen to improve
Hunter
Valley
region of
Further clinical
coast (
et al.
the state of New
cases were
McCrossin 1986)
1986)
In
of
1988,
Lawrence
serological
was
diagnostic
service
for
IFAT)
and
through
tested.
tests to
1992,
In 1990, at the 22nd Annual SOVE Conference in
derived from a North
1991). In that paper it was reported that many` flat' ticks

had been processed without isolating into culture any
specimens
indicate
2. 7%)
Lyme disease,
patients
to the
CDC
seven
and
clinical
( D. Dickeson
and
spirochaetes that might be B. burgdorferi.
Although
some spirochaete- like organisms had been detected in
in both
less than 1
cultures from engorged ticks, attempts to identify these

organisms as B. burgdorferi, using serological and PCR
for
techniques, were unsuccessful. This present report is an
criteria
patients
the first year of this investigation was presented( Russell
were
Lyme disease
Lyme disease
including
Australia
from 2, 446
showed positive results
possible
percent conformed
outside
Borrelia burgdorferi. From
suspected clinical
Only 66(
importance of the disease.
Mesa, Arizona, U.S. A., a paper on the early progress of
strain ( B31) of
referred with
This would
fluorescent antibody test

linked immunosorbent assay
at
were used with antigens
American
1988
enzyme
tests and disease diagnosis.
Westmead Hospital in
initiated
Sydney, N. S. W.; indirect

ELISA)
serological
allow a more accurate assessment of the public health
the state.
a
Lyme disease
and central coast (
infected
R. Munro,
update
with
the
conclusions
of
the
three- year
investigation.
Presented at the 7th European Annual Meeting of SOVE, Bologna, Italy, August 27, 1992.
2Department of Medical Entomology, University ofS ydney, Westmead Hospital, Westmead, NSW 2145, Australia.
3Department
of
Microbiology,
Liverpool Hospital, Liverpool, NSW 2170, Australia.
78
MATERIALS AND METHODS
DECEMBER, 1993
approximately 11, 000( TABLE 2) were processed for

spirochaete isolation. Each year the collections were
Tick
collections were targeted towards those areas
Lyme disease; these
associated with putative cases of
in the
were all situated
by ` flagging'
were collected
removal
the
coastal
and `
border in the
identified,
dragging,'
animals
in
and
stages of the species were most abundant from late-
by
autumn through to mid- spring, and adults from

mid-spring through early- summer.
from
areas
Victorian
the north to the
No spirochaetes were detected by microscopy in
south.
Isolation
of
unfed(` flat') ticks of any species from natural habitats.
spirochaetes
The ticks
attempted.
N. S. W. Ticks
of
strip
from native and domestic
Queensland border in
dominated by larvae of Ixodes holocyclus Neumann

during the late- summer and autumn months; nymphal
live
was
Cultures from gut contents of ticks collected from
until processed,
various localities throughout the length of the study area
in 70
by dipping
surface sterilized
twice in distilled
ethanol, washed
from the ticks
were stored
water, and
yielded 94 separate ` isolations' of spirochaete- like
percent
the midgut
objects (
tissues dissected out aseptically. A portion of the midgut contents was examined
by dark field microscopy for
the presence of spirochaetes, and the rest was
inoculated
into BSKII
1984)
incubated
culture media (
at
Johnson
33 C for up
et al.
and
were sought
by
Molecular
objects (
selective
in
purified SLOs in the absence of bacterial contaminants
isolated from
or improve their growth by manipulating culture
filtration from the
cultures
conditions.
All the monoclonal antibodies yielded negative
cultures.
the spirochaete- like
results when tested against the SLOs, but yieldedpositive
from the ticks
results against controls of the North American ( B31)
was
reference strain of B. burgdorferi. While some positive
monoclonal and polyclonal antibodies and
results were obtained by IFAT using polyclonal
the polymerase chain reaction ( PCR).
isolates'
isolations' were in association with other bacterial
variety of immunochemical techniques
attempted with a
including
of spirochaete- like objects
characterization of
SLOs) found in
and Warburton, Haemaphysalis longicornis Neumann,

Amblyomma moreliae ( L. Koch). All these
and
contaminants, and it was not possible to subculture
constituents and culture conditions were manipulated
the ticks. Purified isolates
The tick species yielding ` isolates'
Media
to three months.
efforts to enhance growth of organisms
SLOs).
were I. holocyclus, Haemaphysalis bancrofti Nuttall
four polyclonal
were tested against
monoclonal antibodies with
Eighteen
such
antibodies, the results were both variable and inconsistent
and three
for the 18 SLOs tested; all the polyclonals tested positive
IFAT. The four polyclonals
with the control.
burgdorferi-FITC, rabbit polyclonal
PCR amplification products of the size expected for
anti- P39 serum, rabbit polyclonal anti- P22- A serum,
the OspA gene were obtained from just one` isolate' out
and rabbit anti- B. burgdorferi whole serum.
The mono-
of 30 examined. In order to confirm the specificity of
against the outer surface proteins
the PCR products obtained, the DNA fragment was

hybridized to a specific oligonucleotide probe under
were rabbit anti- B.
clonals were
OspA
and
used to
directed
OspB,
and
flagellin
detect Borrelia
proteins.
specific
The PCR
DNA in
was
purified and
high stringency conditions. DNA from the SLO cultures
SLOs. The primary targets

the OspA, the flagellin gene
amplified with Fla gene specific oligonucleotide primers
non- purified cultures of the
for PCR detection
Fla), and the 16S

Strain 297
of B.
were
ribosomal gene(
burgdorferi(
Persing et al. 1990).
a North
was used as a positive control;
experiment
in
controls
probe specific to the Fla gene even under low stringency
DNA from Leptospira
conditions. Similarly, DNA from 45` isolates' amplified
were
also run
in
each
to detect any non- specific
order
yielded no products
specific to
B. burgdorferi.
The SLOs were examined by both light and electron

microscopy. Under dark field microscopy the SLOs
for testing for Borrelia by PCR;
appeared straight, rigid, and uniformly coiled( Fig. 1).

They varied in length from 10 m to 300 m, and had
from 2 to 40 complete coils. All appeared to be non-
of the ticks was selected
the target DNA for PCR amplification was the Borrelia

specific
using oligonucleotide primers specific to the 16S

ribosomal gene
Electron microscopy was also

SLOs from culture media. A subsample
amplification products.
used to examine
isolates' which did nothybridize with an oligonucleotide
American isolate)
interrogansserovarcopenhageni was used as a negative

control; " no- DNA"
yielded non- specific amplification products in some
41kDa flagellin
gene(
Persing
et al.
1990).
motile. Electron micrographs showed that these SLOs

RESULTS
Between
January
1990
20, 000 ticks ( TABLE 1)
and
had no distinct cellular structure but were composed of

December 1992,
were
collected
and
fiber-like subunits( Fig. 2) and were not spirochaetes.

Approximately 1, 000 ticks collected from different
areas within the
study
region were
tested using PCR
DECEMBER, 1993
79
TABLE 1. Numbers of ticks collected for spirochaete isolation, January 1990- December 1992.
Species
Larvae
Nymphs
Males
Females
TOTAL
Amblyomma
moreliae
16
41
12
69
A. triguttatum
17
1633
513
79
148
2373
346
2176
66
1681
4269
7420
2057
561
2117
12155
66
46
10
77
199
175
68
245
1033
1033
22
31
10677
4880
784
4062
20403
Aponomma
concolor
Haemaphysalis
bancrofti
H. longicornis
Ixodes
cornuatus
I.fecialis
I. holocyclus
I. tasmani
1. trichosuri
Ixodes
species
Rhipicephalus
sanguineus
TOTAL
TABLE 3). There were
no amplification products that
would suggest the presence of
Borrelia.
In
alone cannot be used as an indicator of Lyme disease.
contrast,
False positives may be obtained due to cross reactions
consisting of naturally infected Ixodes dammini

Spielman, Clifford, Piesman, and Corwin ( recently
with other bacteria, notably spirochaetes. Laboratory

confirmation by culture of the causative organism from
controls
relegated to
Oliver
et al.
synonymy with Ixodes scapularis Say by

1993), from Westchester Co., New York,
U.S. A., yielded positive results under the test conditions.
a local patient has not been achieved in Australia.
Thus, B. burgdorferi has not yet been isolated from
any patient( without a travel history to an endemic area

in the northern hemisphere), animal host, or tick vector
DISCUSSION
in Australia, and there is no definite evidence that B.

burgdorferi s. s. exists in Australia. In the present
The clinical and serological determination
disease
outside
known
and controversial.
of
endemic areas
is
of Lyme
investigation, the SLOs found in cultures from the field
problematical
collected ticks appeared to be cultural artifacts when
In an area without defmitive evidence
infection( i. e. isolation
vectors, vertebrate
hosts,
of causative organism
or
humans),
positive
from
serology
examined by electron microscopy, and the few positive

results obtained with molecular analysis arenon- specific.
It is recognized that the
monoclonal antibodies and PCR
80
DECEMBER, 1993
TABLE 2. Total number of ticks dissected for spirochaete isolation, January 1990- December 1992.
Species
Larvae
Nymphs
Males
Females
TOTAL
Amblyomma
0
14
554
265
67
71
957
232
914
18
748
1912
4862
1115
411
964
7352
I. tasmani
41
29
31
105
I. trichosuri
13
46
60
529
529
18
25
6231
2379
528
1832
10970
moreliae
A. triguttatum
Aponomma
concolor
Haemaphysalis
bancrofti
H. longicornis
Ixodes
cornuatus
I.fecialis
I. holocyclus
Ixodes
species
Rhipicephalus
sanguineus
TOTAL
primers used in this
study may not have been appropriate
to detect an indigenous Australian spirochaete but,

this
notwithstanding
ultimately
spirochaetes.
The
these Australian
of similar
by
shown
microscopy
were
descriptions and light micrographs of those isolates
be
suggest they are identical with our SLOs and therefore

may similarly prove to be cultural artifacts.
not to
electron microscope photographs of
SLOs are comparable with photographs
SLOs
from
There are some major ecological differences
in
between the ecology of Lyme disease in northern
consisted of aggregations of
hemisphere endemic areas and the possible situation in
recovered
Missouri, U. S. A., that

bacterial
the SLOs
concession,
electron
and SDS- PAGE analyses( Alpers 1992, Wills and Barry

1991).
However, the published light microscopy
cultures of ticks
flagellae thought to
originate
from
Australia.
In the northern hemisphere, all principal
contaminating bacteria in the cultures( Miles et al. 1992;
vectors to humans are members of the Ixodes persulcatus
Miles,
group
Similar
in
elsewhere
of ticks( Barbour and Fish 1993). Of the ticks in

Australia associated with putative human infection,
to
I. holocyclus is the logical candidate vector. This species
to be the probable cause of Lyme
has a wide host range and is the most common tick
in Australia
B. burgdorferi
and
from
workers
pers. comm.).
objects
cultures
dissected bloodfed ticks taken from animals by

are claimed to
be
related
disease in Australia, based on serological characterization
species
biting humans along the east coast of Australia.
81
DECEMBER, 1993
4.
fir
r4.,
tl"
JO/
461.
Figure 1.
t.
Dark field micrograph of spirochaete-like objects( SLOs) detected in cultures from ticks, showing the
typically rigid and straight appearance in various lengths, and the uniform coiling( x400).
sc
et,.
f
j'
1` {
4.
s
A
t
k
gyp
',
Zit= ,
a
liti,
r'tdpitt.
Figure 2.
r,
Electron micrograph of portion of one of the spirochaete- like objects ( SLOs) in cultures from ticks,
showing
an apparent composition of
fiber-like
subunits( x5000).
82
DECEMBER, 1993
TABLE 3. Number of ticks examined by PCR for the presence of Borrelia, January 1990- December 1992.
Species
Larvae
Nymphs
Males
Females
TOTAL
Amblyomma
0
119
135
159
137
296
I.fecialis
I. holocyclus
279
53
236
570
I. tasmani
13
19
I. trichosuri
13
13
Ixodes
576
62
396
1038
moreliae
A. triguttatum
Aponomma
concolor
Haemaphysalis
bancrofti
H. longicornis
Ixodes
cornuatus
species
Rhipicephalus
sanguineus
TOTAL
However
tick in
neither this species, nor
Australia, belongs to
ricinuscomplex. Ixodes
the
any other species of

Ixodes persulcatus/
holocyclus is unable to
mission in Europe, northern Asia, and north America is

present in Australia. There are reports of spirochaetes
B. burgdorferi
in our native animals, and it is entirely possible that

some marsupials could be acting as a reservoir host for
Piesman and Stone 1991) but there is no information on
an indigenous spirochaete that through a tick vector
its ability to transmit European strains, and its association
might occasionally infect humans and produce a clinical
or
transmit a North American
with
possible
Australian
strain of
spirochaetes
maintain
remains
unresolved.
Any
consideration
B. burgdorferi
should take
into
or a
of
similar
account
reservoir
spirochaete
hosts
for
in Australia
that the native vertebrate fauna
primarily marsupial mammals) is dissimilar

the Lyme disease endemic regions in the
to that of
northern
syndrome similar to Lyme disease. It was not possible

within the limits of the present investigation to undertake
isolation of spirochaetes from potential animal reservoir
hosts via ear punch biopsies( Sinsky and Piesman 1989);
it is recognized that this is also a way of confirming the

presence ofaBorrelia or similar spirochaete that may be
responsible for the human syndrome in Australia, and
hemisphere( placental mammals), and none of the species
perhaps this could form the basis of future studies.
identified
However, if there is
as reservoir
hosts for B. burgdorferi trans-
local
spirochaete other than
DECEMBER, 1993
B. burgdorferi transmitted
Lyme disease- like
by
ticks and responsible for
in Australia, it has
83
Ramaciotti Foundations.
We are grateful for the
not
assistance of Danielle Avery, David Dickeson, Dr. John
been detected in the approximately 12, 000 ticks

processed during the present investigation and thus
Ellis, Dr. Cheryl Hunt, Joanne Simmons, and Nicolle
might
be
considered
syndrome
to be uncommon or rare.
also due to Dr. Durland Fish of the Mec: ical Entomology
The developmental cycle fort holocyclus generally

to be completed
appears
within one year, with some
the life cycle active
during virtually all months.

The major period of activity for nymphs in southeastern
Australia occurs in the cooler months, and directly
stage of
follows that
Trivett in various aspects of the program. Thanks are
Laboratory, New York Medical College forhis assistance

with our reference field collections in Westchester
County, NY, U.S. A.
the larvae which are most active in
of
REFERENCES CITED
summer and autumn. Nymphs are thought to be the

most
important
stage
for transmission to humans in the
hemisphere because they are more likely to be

infected than larvae, and are much smaller than adult
northern
less
ticks and thus

removed
soon
noticeable
after
and
less
In
attachment.
Alpers, J.
1992.
Borrelia isolated from Australian
ticks. Today' s Life Sci. 4: 40-41.
be
Barbour, A. G. and D. Fish. 1993. The biological and
southeastern
social phenomenon of Lyme disease. Science. 260:
likely
to
Australia, the period of greatest likelihood for human or
1610- 1616.
vertebrate host contact with nymphs ( and therefore

higher risk
appear to
of possible
be
during
infection) in
sylvan areas would
the cool seasons
are most active, although
it is
when
the nymphs
at this time that
human
Lawrence, R. H., R. Bradbury, and J. S. Cullen. 1986.

Lyme disease on the central coast. Med. J. Aust.
145: 364.
activity in the areas is least. During the summer periods,

when
there is the greatest animal and human activity
in tick infested
likely
it is the larval
Johnson, S. E., G. C. Klein, G. P. Schmidt, G. S. Bowen,
least
J. C. Feeley, and T. Schulze. 1984. Lyme disease:
to be able to transmit unless infected trans-
a selective medium for isolation of the suspected
Also, there is little
etiological agent, a spirochete. J. Clin. Microbiol.
ovarially) that
areas,
are most active.
evidence that tick populations are

residential areas of
appears to
be
Australia.
stages (
increasing
in
or near
19: 81- 82.
Instead, the opposite
happening as native hosts are driven from

by urbanization and the introduction of
residential areas
McCrossin, I. 1986. Lyme disease on the NSW south

coast. Med. J. Aust. 144: 724-725.
feral animals and domestic pets, particularly cats and

dogs.
Miles, D., E. Masters, and J. Rawlings. 1992. Isolation
In summary, definite evidence for a spirochaete

causing Lyme disease in Australia is still lacking. While
of spirochaetes in the south- central U.S.
cases of` Lyme
page A30.
disease' have been described clinically
Proc. V
Int. Conf. Lyme Borreliosis. Arlington, U.S. A.,
in Australia, and there is some serological support for a

spirochaetal infection simulating Lyme
disease, aproven
from
Oliver, J. H., M. R. Owsley, H. J. Hutcheson, A. M.

James, C. Chen, W. S. Irby, E. M. Dotson, and D.
K. McLain. 1993. Conspecificity of the ticks
this country. The ecological characteristics of the tick/
Ixodes scapularis and I.dammini( Acari: Ixodidae).
causative organism
has
not
been isolated from patients,
potential tick vectors or possible animal reservoirs
vertebrate
host
cycles
in
southeastern
Australia
are
particularly important in an epidemiological context

and may help to explain why the so- called ` Lyme
disease' in Australia is at best an
uncommon,
if not rare,
J. Med. Ent. 30: 54- 63.
Piesman, J. and B. F. Stone. 1991. Vector competence
of the Australian paralysis tick, Ixodes holocyclus,

for the Lyme disease spirochaete Borrelia
phenomenon.
burgdorferi. Int. J. Parasitol. 21: 109- 111.

Acknowledgments
Persing, D. H., S. R. Telford III, P. N. Rys, D. E. Dodge,

T. J. White, S. E. Malawista, and A. Spielman.
The investigation
grant
was
from the National Health
Council
of
Australia,
and
a research
1990. Detection of Borrelia burgdorferi DNA in
Medical Research
museum specimens of Ixodes dammini ticks.
supported
and
by
grant- in- aid
from the
Science. 249: 1420- 1423.
84
1991.
Russell, R. C.
The Lyme disease
situation
in
Australia. Bull. Soc. Vect. Ecol. 16: 227- 229.
DECEMBER, 1993
method for detection and isolation of Borrelia
burgdorferi from rodents. J. Clin. Microbiol. 27:

1723- 27.
Sigal, L. H.
1988.
borreliosis. Clin.
Sigal, L. H.
and
Lyme disease: a world-wide
Exp.
Rheumatol. 6: 411- 421.
A. S. Curran. 1991. Lyme disease:
Stewart, A. J., J. Glass, A. Patel, G. Wyatt, A. Cripps,

a
and R. Clancy. 1982. Lyme arthritis in the Hunter

Valley. Med. J. Austral. 1: 139.
multifocal worldwide epidemic. Ann. Rev. Publ.

Hlth. 12: 85- 109.
Wills, M. C. and R. D. Barry. 1991. Detecting the cause

of Lyme disease in Australia. Med. J. Austral. 155:
Sinsky, R. J.
and
J. Piesman. 1989. Ear
punch
biopsy
275.
DECEMBER, 1993
THE ROLE OF ECOLOGY AND ECOLOGICAL
MODELING IN VECTOR CONTROL'
M. W. Service2
ABSTRACT: The main issue of this paper is whether we need more ecological data for developing control strategies
aimed at vector-borne diseases. Some argue that we already have sufficient knowledge of vectors, but need more
and better trained vector control specialists. In contrast, others believe our failure to control many vector-borne
diseases is due to insufficient understanding of vector ecology. With the prospect of decreasing budgets the question
is, should research be concentrated on topics that appear relevant to control, or is there still room for academic
may have" spin- offs" relevant to control? With some diseases, like dengue, we may already have
sufficient information on the vectors to decide on the best control approach, which in this case is source reduction.
research that
With other diseases, such as malaria and leishmaniasis more research on the vectors is needed to identify the most
appropriate control methods. Mathematical models concerning vector-borne disease control date back to Ross' s
1911) malaria model, but have they proved useful in helping design better control operations? A problem seems to
be that models have often promised more than they have delivered, and as a consequence models tend to be regarded
with suspicion by field entomologists. However, more recently, by unraveling the complexities of species
interactions, some models have provided valuable insights into population ecology of vectors and appear to have
been successful in identifying the best approaches for control. Nevertheless, despite the proliferation of models, it
must be admitted that as yet they have not often led to the successful implementation of better control. However,
despite these criticisms, I believe modelers and ecologists should be encouraged to work together to try and build
more realistic models that help both vector control specialists and epidemiologists.
relation to their behavior and habitats, and iv) the
WHAT IS ECOLOGY?
scientific study of the interactions that determine the

distribution and abundance of organisms. I prefer the
When I chose the title of this talk I thought it would
last two definitions because they emphasize that ecology
but I
It has
is concerned with relationships between organisms.
describe
be easy to
write the text,
difficult to define the
was mistaken.
let
big
to be adequately covered in this presentation. All I
Ecology is a central discipline in biology and overlaps

others like evolution, physiology, genetics, and behavior.
It is the overlap with behavior that I find particularly ill-
can
hope to do is identify
defined. For example, are studies on dispersal and
proved
its
role, past or
future, in
subject,
vector control.
my
own
mostly
interests.
Firstly,
what
Ecology
is too
few generalizations, and give
some examples of the use of ecological

programs,
alone
on mosquitoes, that
data in
probably
control
reflect
exophily behavioral or ecological? Whatever the answer,

I have largely ignored these issues in this presentation
and concentrated more on population ecology and
is ecology,
apart that
is from
having
modeling.
Colinaux 1973).
Ecologists have for many years studied predator-
There are many branches of ecology, for example books
prey relationships, but more rarely parasite- host

interactions, a topic that more usually occupies the
been described
as" a pleasant science"?(
have been
written on population ecology, behavioral

ecology, evolutionary ecology, theoretical ecology,
community ecology,
ecology,
parasite
ecology,
mathematical
Ecology has
scientific natural history,
and even on mosquito
ecology.
variously been described as i)

ii) the scientific study of the distribution and abundance
of organisms, iii) the study of animals and plants in
minds of parasitologists or epidemiologists. B ut I would
argue that epidemiology is just another, albeit rather

insular, branch of ecology that is concerned with the
interactions and dynamics of infections and people.
Certain concepts are relevant to both ecology and

epidemiology, such as the heterogeneous dispersion of
1Paper presented at the 1st International Congress of Vector Ecology, San Diego, California, USA, 4 October 1993.
2Liverpool School of Tropical Medicine, Vector Biology and Control, Pembroke Place, Liverpool L3 5QA,
ENGLAND.
86
in the
organisms, as reflected
of diseases
in host
distribution
ecological studies are generally regarded as superfluous.
heterogeneity
Only when control operations begin to falter is there

perceived to be a need for vector ecology and behavioral
nonrandom
populations, as well as
in their genetic makeup or behavior. A particular
be
systems
reporting
frequently
extend
is
studies. Then, however, because of the lack of precontrol biological data, it is difficult to decide whether
many years, and disease

be linked to changes in human
or not control programs have caused changes in behavior,
size
demographic data.
cynics would argue that any control scheme has built-in
interest,
obsolescence and that sooner or later control will decline,
and
other
Nevertheless, despite
and
ecology
little from
envied
by
ecologists,
over
prevalence can often

population
strength
formonitoring disease prevalence
of epidemiology, and one to
that
such
similarities
of
epidemiology have, until recently, benefited
cross
DECEMBER, 1993
fertilization.
such as increased exophily of malaria vectors. Some
and ecologists will be asked to explain why.

Tsetse flies provide a classical example of the
importance of ecology in control, where as early as the

DO WE NEED ECOLOGY?
Before 1940,
modern
biological
simple
only
chemical
limited
it was believed that control of sleeping sickness had to

be based on a sound knowledge of vector ecology. Later
based on environmental
in the 1940s and 1950s, ecological studies identified the
few
resting sites of tsetse flies on vegetation, which led to the
methods,
Such
agents.
amount of
virtually
pesticides
unknown and vectorcontrol was
manipulation,
1920s detailed ecological studies were initiated because
were
and use of a
approaches
required
biological knowledge,
such as
larval ecology including larval feeding

behavior. In 1927, the League of Nations' Malaria
mosquito
Commission
emphasized the need for research by stating

The only prospect of real progress lies in renewed
activity in the continuous study of the disease in all its
Anon 1927). F. L. Soper appreciated
aspects"(
spraying of these sites with residual insecticides.
Lumsden( 1962), however, was very critical of the value

of ecological studies, believing that much research had
been academic and that the entomologist had" had his
day," because his research had failed to generate

economic and reliable vector control strategies. He
foresaw the future in new drugs and vaccines. Now
the need
thirty years later, prospects of controlling human or
for ecological knowledge, and his successfulAnopheles
animal trypanosomiasis with vaccines or drugs seem
in Brazil ( 1934- 1940)
remote. During the 1970s, effective, cheap and simple

traps were developed by ecologists for studying tsetse
gambiae eradication campaign
Soper and Wilson 1943)
was
based
on a comprehensive
understanding of the ecology of this vector.

With the advent of the DDT-era in the 1950s
behavior and today these traps, insecticide-impregnated

and the
or otherwise, together with insecticidal screens, offer
euphoria of its early years,
vectorecology was considered

This belief is supported by the
the best means of controlling sleeping sickness. So, as
unnecessary luxury.
declaration in a 1960 issue
Jordan( 1986) has said, " perhaps the entomologist has
an
by
is
eradication
above
administrative problem,
need to
of
World Health
published
World Health Organization that " Malaria
the
know
about
WHO 1960). This
its
all
exophily,
and
inadequate,
and
tools for malaria control, and thus a need to develop
almost all we
better ones. In contrast, Gratz( 1985) considered that the
scientific and technical aspects"
most urgent need was not for more research on vectors
or better control strategies, but for more well-trained
resistance problems and a
vector control specialists. Similarly, Reeves( 1982)
that our knowledge on dispersal,
speculated that the failure to control dengue, yellow
survival
and this
organizational
confidence soon came to an end.
There followed insecticide

gradual realization
an
for we now know
not after all had his day."

Bradley ( 1991) believed that we had imperfect
rates
forced
of
vectors
was
often
fever, and other arboviral diseases was, in part, due to
renewed attention to vector
lack of government commitments and an acute shortage
ecology and applied research. It is interesting to recall

that at its second meeting in 1979, the UNDP/ World
of well- trained vector specialists. However, he also
Bank/ WHO Special Programme for Research
Training
priorities
in Tropical Diseases listed among its research

for malaria control" vector biology, ecology,
behaviour,
despite
some
and
and control" (
past
health
WHO 1979). Nevertheless,
lessons there is
still a
lack
of
interest
by
we do not need more ecological knowledge for control
is a contentious viewpoint. However, it may be true with

a disease like dengue, where efficient source reduction
authorities or others who control the purse
linked with willing community participation can be
for allocating resources for ecological research

vectors before control programs are initiated.
effective, although getting and sustaining community

cooperation is an elusive goal( Service 1993a).
strings
on
argued for more research, pointing out that markrecapture techniques had been invaluable in elucidating
many relevant aspects of vector behavior. Saying that
Furthermore,
as
long
as
control
remains
effective,
Malaria
control campaigns
in the
past, such as
DECEMBER, 1993
during
the
building
development
limited
ecological
epidemiological
sufficient
the Panama Canal and the
of
the Tennessee
of
Valley,
information
was essential in order to pinpoint breeding sites and
succeeded with
focus larvicidal control on these areas. Also in Panama,
and without the use of
Chaniotis et al.( 1982) concluded that more knowledge
because there
largely
models,
87
biological information
was needed on the resting habits of the Lutzomyia
was
about the mosquitoes.
vectors of cutaneous leishmaniasis before control could
that environmental measures,

on a
become feasible. I also think that most of us would agree

with Asman et al.( 1981) that information on the native
understanding of the ecology of Anopheles

quadrimaculatus( Kitron 1987, for references). Malaria
mosquito population is vital for any genetic control

program. They identified ecological aspects, such as
It is
acknowledged
level management,
water
principally
were
based
good
was also eliminated
from the Pontine
in Malaysian
degree of isolation of the target population and the
Italy
marshes of
Assam
strength of densitydependent population regulation as
tea plantations. However, in many situations control
and controlled
activities were restricted to
immediate
and
rubber estates and
It is
1950s very little was known

Simulium neavei, an East
dosing
and
including
streams with
forces
also true that
in the
inoperative. One of the reasons for the failure to sustain
protecting
communities.
onchocerciasis,
the work
key issues. Weknow that immigration and strong densitydependence can make any sterile male release program
biology
egg sterility in Culexquinquefasciatuspopulations near
vector
of
Delhi by the release of sterile-males was because the
it bred. Bush clearing
extent of infiltration of fertilized females from outside
African
where
DDT
the
of
about
the vector and river blindness from Kenya.
because there
possible
blindness
were
dispersed very little.
fragile
quinquefasciatus in the Delhi and Yangon areas, none
Consequently,
was
there
In contrast, ecological
were no problems with reinvasion.
data have
ecological
Success
discrete foci
and the vector occupied a
niche and
of river
the project area had been underestimated. Macdonald

1991) concluded that although an unparalleled amount
of ecological information had been gathered on Cx.
nevertheless eradicated
indispensable in the
of this was sufficient to achieve lasting control in either

India or Myanmar( Burma).
operation of the
Despite Aedes aegypti being such an important and
Ochocerciasis Control Programme( OCP) in West Africa.
common vector, and moreover a container-breeder that
For example,
makes it well- suited to biological investigations, there

have been few good ecological studies on its population
proved
surveys showed that parous, and
potentially infective, adults

be
complex continued to
effective
larvicidal
very
dynamics. For instance, relatively few field-based studies
The big question was, where
on adult survival rates have been undertaken although
coming from?
Careful biochemical
such knowledge is crucial for understanding disease
identified the adults as S. damnosum and Simulium
epidemiology and also of considerable importance in
were the adults

studies
biting despite
caught
control.
therefore
Simulium damnosum
of the
sirbanum(
Townson
and
circumstantial evidence
outside the
OCP
transported
Meredith 1979),
strong
evaluating control programs. In contrast, several mark-
indicated that they were breeding
recapture studies have shown that adults disperse very
in the
area
while
southwest and were
being
prevailing winds into the control area.

Some adults had apparently dispersed some 3001cm and
possibly
on
as much as
damnosum
could
500 km. We
fly long
always
knew that S.
distances, but the
extent of
this had not previously been appreciated. This is a good

example of our
failure to
understand the
before embarking on control.

Although there has been long- term
behavior
of a
vector
rivers and streams
in the OCP
area,
non-
life( Yameogo
et al.
target organisms,
undoubtedly killed, but
there is
Awareness
by
of the disease. However, we must take into account the

geographical
differences
in
their
degree
of
to
some
their
vertical
stratification (
Service 1982). Such
variations in behavior complicate our understanding of

yellow fever epidemiology, and stress the importance of
unraveling the local ecology of vectors if control is to be
introduced. Working in East Africa, Haddow ( 1977)

said of yellow fever that" there has been a tendency to
theirimportanceinregulating
oversimplify the apparently straightforward situation,
S. damnosum
of the need
Petersen
yellow fever, namely Ae. aegypti, Aedes africanus and

Aedes bromeliae, for an epidemiological understanding
very little is known
even now
about natural predators and

populations of the
of
apparently
1988). Nevertheless
including
know all we need know about the African vectors of
anthropophagy, domestication and endophily as well as

larviciding
no evidence of severe adverse ecological changes
aquatic
little, information that is relevant to both epidemiology

and control( Service 1993b forreview). We may feel we
predators,
are
and this has often been an obstacle to progress." Focks
complex.
for
ecological studies
1983),
is
et al.( 1979) provide another example of the lack of data
working in
on Ae. aegypti. When they analyzed attempts to control
understanding of the
ecology of the aquatic stages of Simulium quadrivittatum
Ae. aegypti and other container-breeders in the U. S. A.
provided
Panama believed that
et al. (
a thorough
who
by releasing Toxorhynchites mosquitoes they found that
DECEMBER, 1993
88
factor contributing to lack of success

limited knowledge of the interactions between
Moore ( 1990) also stressed the
the most common
to
was the
importance of ecological and epidemiological research
problem- solving.
prey.
It may be easy to find breeding places of mosquitoes but prove difficult to devise a sampling program
but believed it should be more " solution oriented" as
predators and
opposed to"
intensely academic research." The counter
argument is that it is difficult to know whether
in terms
information obtained during curiosity-driven research
of pupal production, or better still, numbers of emerging
will later be relevant to control. At least that is the
There is little knowledge on the natural mortalities
excuse of many researchers, and it has more than an
identify
that will
adults.
the most productive habitats
by mosquito larvae and even less on the

factors regulating their larval population size. This may
not be an important constraint with larviciding because
experienced
insecticides
EXAMPLES OF ECOLOGICAL STUDIES

RELEVANT TO CONTROL
density- independent
catastrophic
cause
element of truth in it.
mortalities that are unaffected by densitydependent

However, it is
processes.
biological
control
fashionable to
now
this usually
and
advocate
necessitates
an
understanding of the birth and death processes of vector

populations( Rogers 1983; Service 1985). This is because
biological
dependent
introduce
aquatic
processes.
control
life
late
to kill the
processes
compensate
many
data
of
wealth
not
of crucial importance because it determines whether the

infection persists( Ro> 1) or dies out( Ro< 1). The value
of Ro will determine the equilibrium prevalence of an
accumulation of research
infection. Clearly a minimum number, called the critical

or threshold, density( see p. 5) of vectors is required to
control.
sad state of
in
sustain transmission. Disease eradication is about
mosquitoborne arboviruses pointed out that
thresholds or breakpoints. Cuellar( 1969) was an early
by
DeFoliart
This
1987),
et al. (
proliferation of research over the
decades
have
we
profited
little
transmission through
ecologists collected
have
or
control
with regard
vector control.
data that is
workers
last few
biologists
and mathematicians consider
correct.
it
which he referred to as the critical level of interference,
is this?
needed to be reached for species eradication. He
to
presented some theoretical values for malaria vectors,
ecological
but the paper stimulated little interest.
Molecular
Although we may speak about the goal of reducing

vector density to a critical threshold or breakpoint for
transmission to cease, we usually have little idea what
this threshold is, let alone have the means of achieving
remarkable
may not be able to tell them how long a vector

lives, how far it flies, and what it does all day. They feel
when we
have had decades
answers to these
their turn to
of opportunities to
simple questions,
apparently
dazzle, if not blind
us, with
find the
it is
now
biotechnology
and mathematics.
Should
we
continue
to collect more and more
data in the belief that
proponent of establishing a critical vector threshold,
to reducing
Why
ignored
I think that both
are
who
not relevant
studies?
that we
transmission. The basic reproduction rate of a parasite is
and
In fact, there is
despite the
control,
understanding disease epidemiology and determining

the intensity and frequency of control needed to interrupt
spectacular control successes.
affairs was realized
Have
are important in
led to
ecological
1970s has
since the
widening gap between the

data and successful disease
viral
Ro)
is intrinsically capable of producing. Or another way of

expressing it is, that Ro is the average number of
secondary cases of infection that are generated from one
primary case in a susceptible host population of defined
density. It is analogous to the net reproductive rate of an
organism, which is the number of offspring that gain
sexual maturity. A parasite' s basic reproduction rate is
increasing
discussing
rate (
the average number ofsuccessful offspring that aparasite
strong densitydependent larval mortality, then even a

large decrease in adults may not be followed by a
significant reduction in the numbers of emerging adults.
The
reproduction
early stages, because such

the likelihood of densitydependent
mortality
allowing increased survival rates to

for early mortality. Furthermore, if there is
epidemiological
Life-cycle characteristics, especially the parasite' s

basic
stages of
rather than the
reduces
mortality
greatly affected by densityFor example, it is preferable to
control agents are
Critical Thresholds
it. There have been several unsuccessful attempts to
correlate mosquito densities in light traps with cases of

WEE
and
SLE ( Eldridge 1987). However, in Kern
County, California, when the mean number of female

Culex tarsalis caught per night in a New Jersey light
know
trap is one or less there is no evidence of WEE
enough, or should we concentrate on ecological studies
transmission, whereas, with a trap index of two to nine
that we judge are relevant to control?
transmission is maintained, albeit at a low level. For
ecological
we can never
Kay ( 1985)
suggested we should think of the potential usefulness of
the data being collected and that they should be oriented
SLE, virus activity is not usually detected when the

nightly catch is below 10, and SLE stops being a
mean
DECEMBER, 1993
even if in a particular study a critical density was found,
Cx. tarsalis densities
it is unlikely to be applicable in others areas because
100 or more per trap night there is often a decrease
different geographical populations of Ae. aegypti can
1990).
rise to
89
index is below 20( Reeves
disease
significant
et al.
when the
Paradoxically, when
in both WEE
SLE. Several hypotheses have been
and
proposed to explain this,
Reeves
processes(
et a1.
vary in their vector competence. Daily survival rates

will also likely differ considerably, both seasonally and
geographically. In Malaysia, however, Cheong( 1986)
including densitydependent
1990). These include increased
attack rates
reported that during 1973- 1982 there was a good
switch
leading to reduced feeding success, and a

away from biting on birds to mammals that are
association between the" Ae. aegyptiindex"presumably
inefficient virus reservoirs, but neither has been proved.
the house indexand the numbers of reported dengue
Olson
infection
and
1979)
et al.(
cases. However, there is usually little or no association
rates, sentinel chicken seroconversion rates,
between entomological population parameters and
human
cases of WEE and
areas
many
light trap data and
mosquito
studied
California
of
SLE
and
between the indices
correlation
over
21
reported
of all
from
years
dengue cases.
Culicineandanopheline- transmittedfilariasisdiffer
a positive
three factors.
in their host- parasite
Serological conversion rates of WEE in sentinel chickens
trap indices were one
peaked when
for SLE.
to ten
to
They also obtained a
apply to the light trap indices
to
five, and were five

conversion
factor
in
urban areas.
Dye ( 1992)
to culicine- vectored
filariasis
and "
facilitation"
( anopheline- transmitted filariasis), in the transmission
interpret the
of lymphatic filariasis. Basically, with limitation the
in
proportion ofingested microfilariae surviving to become
But urban light trap indices
infective larvae decreases as more microfilariae are
to
epidemiological significance of the numbers caught
traps placed
relationships.
summarizes the theory of" limitation," that is applicable
that accompanied viral activity in nearby
rural areas
ingested by the vector( culicines). With facilitation the
low that they failed to provide a sensitive

indicator of expected levels of viral activity( Olson et al.
proportion increases as the number of microfilariae
1979).
number, declining only at greater microfilarial densities
were
so
Despite
densities
cannot
say
densities
some
success
Cx. tarsalis
of
with
any
with
correlating
critical
ingested increases from very low up to an intermediate

(
anophelines). It is claimed that with anophelines when,
with viral outbreaks, we still
because of control efforts or some other reason, the level
great confidence to what critical
ofmicrofilariae is reduced to below a critical density the
be
to prevent viral
disease will tend to die out without further control
transmission to humans. This is not altogether surprising
efforts. This may be a result of the pharyngeal armature
when as
factors
vectors
long
must
1967, Reeves identified
ago as
influencing
less information
reduced
the
of
ecology
some
WEE. We have
50
even
of Anopheles that destroys some ingested microfilariae,

but
which
does
not occur
in
culicines.
Webber and
Culex
Southgate ( 1981) found that in the Solomon Islands
sampling
insensitive. However, Culex
ovitraps as developed by Reiter( 1987)
may help provide
" species No. 1" was below 0.66 hr1, or equivalent to
on the population
threshold of
pipiens, an urban vector of SLE, mainly because

procedures
a reliable
density
have
proved
sampling
for establishing
method
thresholds.
find
no
significant
numbers of adult
any
of the three
Ae.
1. 55 and 2.23 percent there was a threshold level

( breakpoint) below which the disease could not be
in
from houses
and
or even with the numbers
ovitraps.
Similarly
reported
maintained by Anopheles sinensis providing, however,

that no one had a density of more than 12 microfilariae
in Honduras,
per 60 nun-3. Webber( 1991) argues that establishing
there was no simple
such critical densities shows that control should be
and
aimed at reducing microfilarial densities and not trying
resting collections. This is a

limitation because it is the relative size of the
to cure every infected person. The required threshold
in
adult
adult population that
is epidemiologically important
and the most appropriate

of control.
Despite
in evaluating the
transmission
has
because there
measuring
never
microfilarial density can be achieved by either reducing

the vector biting rate or the density of the parasites.
effectiveness
Despite the above studies on threshold densities
the research that has been
Southgate ( 1992) considered that at present it was
threshold level for disease
impossible to define a tolerable level of lymphatic
been clearly defined, probably
filariasis transmission, but concluded that such a tolerable
all
undertaken onAe. aegypti, the
of
et al. (
between larval indices in households
numbers caught
serious
1990)
aegypti caught
Gil-Bellorin ( 1991)
association
in Hubei Province, China Zhang et al.( 1991) found that
relationship between the
larval indices,
of eggs collected
eight bites a night, bancroftian filariasis died out. Later
when the prevalence of Brugia malayi was between
In the Dominican Republic, Tidwell

could
critical
when the critical biting threshold of Anopheles farauti
are still no efficient and reliable methods

adult population
densities.
Moreover,
level of transmission of Wuchereria bancrofti will

probably be
much
lower than the levels necessary to
DECEMBER, 1993
90
incidence
an
produce
transmission
of new
Therefore,
microfilaraemia.
intensity
be
must
in
and control must result
cases
than that needed to produce a zero
Wuchereria
know
prevalence rates and epidemiological equilibrium in
Dye( 1992)
Gambia. Even if critical vector or parasite densities are
greater
with
Onchocerca
by
West Africa
level
humidity, and the extent of ground water can play a

crucial role in changing the threshold, as Reiter( 1988)
described for the encephalitis viruses in the U.S. A. In
addition, there may be physiological factors, such as
exceeding 100
geographical variations in vector competence and
yearly( Annual
ATP), has been arbitrarily
parasite strains with different host virulence, which
deposition
volvulus
the
obtained, environmental factors such as temperature,
to
needed
Annual
larvae
Transmission Potential chosen
be
and
exceeding 1000 Simulium

Biting Rate- ABR)
not
per person per year(
combined
will
Brugia
apparently homogeneous clusters of villages in The
really is abreakpoint for eradication.
density
critical
of
analysis
dynamics
population
whether there
A
bites
critical
more
has described inter- village variations in malaria
efficiency
incidence rate of new
of
cases, or new episodes of microfilaraemia.
believes
within very small areas as well, leading to the term

microepidemiology. For example, Greenwood( 1989)
for measuring
and improved,
refined
level
detectable
with
methods
not
on a person
make it extremely difficult to apply threshold densities

over a wide area. However, the costs of getting local
information on the microepidemiology of a disease can
Onchocerciasis Control Programme in
as the threshold needed
of control of river
for
blindness to be
an acceptable
attained.
These
be prohibitive.
two parameters will function as a single criterion because

be
neither of them alone will
sufficient to
define
WHO NEEDS MODELS?
an
acceptable risk of transmission. In 1977, WHO proposed

that settlement of areas previously ravaged
blindness
can still
lower ATP there
modeling of vector-borne diseases began with the
The
project area
epidemiological malaria models of Ross, who in 1904
1, 235, 000 km2, but no provisions have been
made
presented a paper entitled " The logical basis of the
for any
maintained
However,
some
now covers
General Considerations
river
Modeling has been more widely applied to malaria

than to any other vector-borne disease. Mathematical
years.
be
by
ABR
at
ATP have been

least two
be allowed only
should
level
after the
below these levels for
even with a
of transmission.
geographical variations
in establishing the ABR.

Dietz( 1988a) pointed
and
in biting intensities
sanitary
policy
of
mosquito
reduction"
to
the
International Congress of Arts and Science at the

Universal Exposition in St. Louis. In 1908 Ross
out that malaria models tend
to focus on a threshold value of the
capacity
itself( De Zoysa
developed a set of differential equations to explain the
Gramiccia, 1980). However,
mathematical analysis of the laws that regulate the
this has been exaggerated.
in
amount of malaria in a community presented

subsequently( Ross 1911). Little attention was paid to
may lead to eradication, although this

isolated area of transmission without any
mathematical modeling of malaria transmission until

the 1950s, when Macdonald( 1956) showed that a small
immigration
of infected vectors or
humans. Even if this
reduction in the survival rate of adult anopheline vectors,
improbable
situation
the reduction in vector
as caused by intradomiciliary spraying, could

dramatically reduce transmission. Since then, the malaria
below
which malaria cannot maintain
1991; Molineaux
et al.
vectorial
the practical
and
importance
This is because it
of
suggests that a partial reduction
dynamics of malaria in Mauritius, with a fuller
biting density
relies on an
densities
be
required
in
exists,
areas of
intense transmission
can
low as one mosquito bite every 20 days!

It is becoming increasingly obvious that the ecology
as
of disease transmission can
model has been refined, culminating in Dietz' s model

1988b), which takes into account densitydependent
vary considerably in different

country. Consequently, baseline data that may
have been collected in an area prior to control may not
effects in regulating transmission, a concept that had

previously been largely ignored. Such a model,
incorporating density-dependent regulation both in
be widely applicable. For example, malariaepidemiology
vectors and humans, shows that a reduction in the
has been
over
malaria parasite' s basic reproduction rate in the host is
but relatively little is known about vector

and
transmission elsewhere in the country. So,
ecology
unfortunately, instead of planning control for large
more effective in reducing transmission than a reduction
parts of a
60
studied
in the Kisumu
area of
Kenya for
years,
geographic
areas,
developing
measures
ecological situations, and this
Moreover, there may be
becomes clear that a sporozoite vaccine would be more
to
effective than a gametocyte vaccine ( Dietz 1988b).
tailored to specific
However, such recommendations may be divorced from

reality because the search for a malaria vaccine has
consideration
that are
be
in parasite reproduction in the vector. From this it
must
given
is much more demanding.
epidemiological
differences
shifted
away from the
sporozoite to concentrate on the
DECEMBER, 1993
91
Cox 1992).
aquatic collections, such as rice fields, and in these
hyperendemic
Nigeria, about
situations it is not easy to understand what is going on.

Ecologists have traditionally studied a single species in
98 percent of the population would have to be immunized
a well defined area, but Anderson( 1991) has argued that
giving life- long protection
the ecological community must" think big" and try to
pre- erythrocytic or erythrocytic stages(
has indicated that in
Modeling
in the Garki
area, such as
area of northern
soon afterbirth with a vaccine
to achieve malaria eradication ( Anderson and
1991).
This
is virtually impossible to
May
interpret population changes of pests over much larger
achieve,
late
areas, making full use of new technology, such as

remote sensing. Another problem is that although in the
past some relatively long- term studies have been
1960s, the disease is still amajorcause of child mortality.
undertaken on some organisms, this type of approach is
It
becoming less popular because funding bodies want
when we recall
especially
a
that although there has been
and effective measles vaccine since the
cheap
can
aim
be
however, that it is
argued,
develop
vaccine
that is
immunity
partial
giving
more pragmatic to
delivered only to people most at risk of malaria mortality

or
morbidity, such as young

women. In other words
develop
northern
diversities and complexities of field situations, and this
an anti- disease vaccine
leads field workers to declare that mathematical model-
making is nothing but a game played by those who have
malaria control
Nigeria,
computer
Molineaux
on
instance,
of
basic
chemotherapy
generalizations and rather wild claims in order to produce
and promote mathematically elegant models. Public
of malaria epidemiology
Gramiccia 1980). In describing the
the project, Najera ( 1974) put the
health workers realize we live in an infinitely complex
baseline data. There
of
discrepancies between
reproduction
rate
uncommon problem, and
of
is
world and this induces skepticism and modeling falls

into disrepute. Another problem is the mathematical
were,
for
complexity of many models that non- mathematicians
estimates of
the
fmd difficult to understand. It is also true that theory has
derived from
malaria
data. This is
often caused
reliable values of such
getting
often a tendency for modelers to make over- sweeping
Macdonald'
on
epidemiological and entomological
of
and
the aid of
s
the absence
wide
no access to the facts. It has to be admitted that there is
model
and
shortcomings
blame
based
in
research project
and was planned with
simulations
mathematical
field
employed mass
DDT house- spraying
Most modelers have little or no experience with the
and pregnant
children
and not an anti- malaria one.
The Garki
quick" fixes" for their investments.
tended to flourish at the expense of good data collection,
not an
and predictions have not always been sufficiently
by difficulties
compared with observations. For instance, Becker( 1979)
seemingly
simple
found that
of
75
publications (
1974 to 1978) on
biting rate, the daily survival rate

of vectors, and also the impossibility of
knowing the
proportion of sporozoite- infected mosquitoes biting that
mathematical epidemiology only five had any reference
actually cause an episode of malaria. Dye and Hasibeder

1985, 1986) discuss the importance of allowing for
mathematical models was likely because they had not

fulfilled the expectations they had raised a decade or so
parameters as the man-
in host
heterogeneity
epidemiological
models.
Hasibeder 1986) to
by
the
selection
They
by
Dye
in
that the reluctance of field malariologists to apply
earlier.
and
A counter argument is that even simple models may
predictions
be able to unravel the complexities ofspecies interactions
wondered (
what extent control
Garki
vectors
to empirical observations. Bruce-Chwatt( 1976) believed
Molineaux
and
and help focus attention on the most important biological
Gramiccia 1980) were misleading because the possibility
parameters that govern transmission and on the best
generated
biting
malaria model(
ignored. Such
heterogeneity
in behavior appears to be common in biology. A readable
strategies, at least in theory, for control. For example,

although acknowledging that there was often a lack of
modeling applicable to malaria

is given by( Dietz 1988a).
relevant and reliable ecological data that can be used by
a surfeit of ecological publications
that modeling had provided valuable insight into the
of nonrandom
was
account of mathematical
transmission and control
There has been

on
medically important
arthropods over the
last 20
modelers,
population
Reeves
et al. (
ecology
1990) nevertheless believed
and control of
Cx. tarsalis.
For
instance, simulation models had identified the best
increasingly
mathematically gifted biologists have

focused their attention on modeling vector
populations.
In contrast, ecologists have tended to study
importance of adult survival rates and temperature on
occupy discrete habitats such as containerbreeding mosquitoes because their biology lends them
vector efficiency.
Despite all the above criticisms and constraints, I
years,
and
animals that
to
having
several
their populations censured and tracked over
generations.
Many
of the
important
mosquitoes colonize more complex and
times for insecticidal control and identified the
believe modeling can provide a framework for
vector
considering the advantages and disadvantages of
heterogeneous
different types ofcontrol and identify the most susceptible
92
biology
aspects of a vector' s
Simulation
on which to aim control.
have clearly
identified adult vector survival rates as the most important
single
factor affecting
Reducing
for
experiments,
survival
example,
infection
population
rates
rates.
logarithmically
can
transmission potential. Koella (
1991)
reduce
gives
very
DECEMBER, 1993
temperatures.
In Louisiana, Focks
et al. (
1988a, 19886) built a
life- table model of the population dynamics of
Psorophora columbiae breeding in rice fields. By

integrating information on weather, host densities( e.g.
cattle), rice hectarage and practices, and any abatement
readable account of the use of mathematical models,
practices, the model successfully predicted the effect on
he restricts his discussion to malaria models,
populations of spraying cattle with insecticides( Focks

et al. 1991) and treating them with ivermectin ( Focks
and although
many
of the points
he
raises are of much wider
interest.
1991). The model can suggest the most appropriate

Examples
of
Modeling
frequency of control operations and therefore deserves
Rochet ( 1990) using field- collected data from
Pondicherry, including
Das ( 1987), built
those presented by Rajagopalan
deterministic
models
for
general,
further evaluation.
In Australia, simulation modeling has suggested
her
that Murray Valley virus is maintained as an endemic

infection in southern parts of the country without human
simulation models generated estimated values that were
involvement, and that periodic introductions from the
in
data,
north are unlikely ( Kay et al. 1987). In this instance,
example, the
models were not used to estimate biological parameters,
infected
such as inoculation rates or to provide estimates of
and
simple
filariasis transmission. In
the dynamics of
good agreement with observations and actual
except
for
mosquito
infection
rates.
For
model overestimated the mean proportion of
in
mosquitoes
underestimated
Various
an
area
it in
under
explanations
discrepancies,
lacking
an area
including
but
critical vectordensities below which transmission ceases,
vector control.
but to provide a theory of how the disease is maintained

and transmitted, based on a retrospective study of past
epidemics. The next step after such an approach is to
control
vector
were
for
offered
heterogeneity
possible
these
in the
susceptibility of Culex quinquefasciatus to infection

with Wuchereria bancrofti and the oversimplification
of
the exponential survival
model
for
used
adult
mosquitoes. The model suggested that reducing mosquito
emergence, such as
would
be the
by source
most effective measure
filariasis transmission. The
create one patent

reported values
infection,
is
which
in filariasis
for
larviciding,
interrupting
model also showed
150,000 infective bites
mean of some
Because
reduction or
that a
was required to
In Thailand, Southwood et al. ( 1972) counted the

numbers of eggs, larvae, and pupae of Ae. aegypti in
water-storage jars left uncovered for 48- hour periods.
From such data they constructed life-tables and

concluded that mortality between eggs and 2nd-instar
larvae was the most important in regulating population
size and was densitydependent.
Rogers ( 1983),
similar to previous
however, reanalyzed their data and concluded that the

greatest population loss was from time of pupation to
studies.
Cx. tarsalis in
oviposition, and that mortality from egg hatching to
California as a vector of both SLE and WEE, there have
2nd- instar larvae was the next most important
been
contribution to total generation mortality. Dye( 1984)

also reanalyzed their data and found that densitydependent mortality acted on the eggs, but mostly on the
the
of
importance
obtain supporting data from the field.
several attempts to
predict population
strategies(
develop
of
computer models
densities and simulate various control
Greever and Georghiou 1979). However, it is
debatable
whether
any
effective
control.
Greever
model
has
and
resulted
in
more
adults, and emphasized that accurate predictions of
Georghiou ( 1979)
population size in models demands reliable information
For
on densitydependent mortality. These examples
recognized that their model was oversimplistic.
instance, it did
vectors, and
data
not take
they
into
account the migration of
also pointed out the
they had
inadequacy of the
illustrate that even those engaged in population modeling

may disagree on the interpretation of data
to build their model.
Rogers( 1983) also postulated that the introduction
offered complete printouts of their simulation
of control measures that caused 90 percent mortality of
available on which
They
to
program on request and

model.
invited
Georghiou ( person.
others to
comm.
improve
1993)
their
adults would give negligible mortality in the next
admits that
generation because any reduction in eggs laid would be

compensated for by a similar reduction in density-
they received only two requests forprintouts. Preliminary

studies
in California
appear
to show that years with
dependent larval mortalities. However, if the same
highest levels of WEE activity were those with the latest
degree of mortality was aimed at the larvae, then good
dates for the
80 F
control would be expected. In other words, larviciding
Milby 1992). Thus, it seems possible to predict this

intensity of WEE transmission from early spring
and source reduction are the best tactics, whereas
accumulation of 50
degree days
over
insecticidal
fogging and ULV applications to kill adults,
DECEMBER, 1993
BULL SOC. VECTOR ECOL
have been
93
the 1960s,
observations which were thought to be caused by
be ineffective in reducing Ae. aegypti populations.

Nelson( 1991) also concluded from modeling that ULV
unaccounted for variations in some of the biological
methods which
advocated since
will
spraying
have little
likely
will
long- term
effect
parameters. This again stresses the difficulties ofworking

with biological variables.
on
densities. This is not to say that efficient

adulticiding will fail to stop a dengue epidemic, but the
point is that ifAe. aegypti populations are to be maintained
reducing
These examples of modeling demonstrate that if
adult
realistic information is used, such as actual data or at

least good estimates, then models can be meaningful. In
low levelsthus minimizing the risk of epidemics

then larval control is most appropriate. In fact, many
marked contrast, an earlier model of Dietz ( 1988c)
field
years this would be followed by recrudescences of

onchocerciasis. However, it is generally accepted that
at
have been saying this for
workers
Gubler ( 1991) recently

effective
It is
way to
control
stressed that " the
dengue is
predicted that even if the OCP extended control over 20
some time, and
only truly
when he built his model the field data he used was
source reduction."
to note that modeling identified
interesting
key meteorological
inappropriate,
so
his
conclusions were
wrong!
More
factor governing
recently, using computer simulation methods, Plaisier
the dynamics ofAe. aegypti populations in New Orleans
et al.( 1991) predicted that in the absence of immigration
temperature as the
Portier
1985). In
contrast, temperature was not
either by infected flies or humans, effective vector
in predicting larval abundance, but

in Puerto Rico. This led Moore( 1985) to
control for 14 years would reduce the risks of
et al.
found to be
useful
rainfall was,
postulate that regression models
based
on rainfall
recrudescence to less than 1 percent.
data
Birley
and
Davies ( 1984), using simulation
can predict outbreaks of Ae. aegypti populations and so
modeling, concluded that under certain circumstances,
decisions. He
such as cooler river water, the interval between dosing
be less costly
water with insecticides could be extended from seven
provide
a means of
control
making
considered that such an approach might
than continual labor- intensive
larval
The OCP in West Africa is the
biggest
world' s
vector control operation, and the amount of international

parasitological, medical, epidemiological,
expertise
entomological, and social

that
far
has been
put
into this
for any other vector- borne

disease project. The program has generated more applied
program
days to 10- 14 days. Working with a modified simulation

model of Birley and Davies( 1984), Cheke and Walter
surveys.
surpasses that
any other similar control program, so it is

surprising that mathematicians have been attracted
the challenge of creating models to both quantify and
1988) confirmed this conclusion. They also found that

in very hot areas it might be necessary to treat rivers at
four-day intervals because of the rapid development of
the vectors. However, these findings, especially those
of Birley and Davies made some nine years ago, have
research than
not led to modification of the intervals between
not
insecticidal applications, despite the fact that longer
by
predict epidemiological
1992, Plaisier
1986) built
et al.
tendencies ( Habbema et al.
1990). For
example,
a catalytic model to
epidemiological trends
during
Remme
et al.
age- specific
study
spraying intervals
Why? It is
undoubtedly because of logistic problems. A weekly
would
save
money.
routine is much easier to follow than one in which

spraying has to be
repeated on
different days.
Also,
a period ofvector control,
there was some resistance to adopting longer intervals
concluding that changes in prevalence rates were too

insensitive to assess the effectiveness of vector control.
because it was considered that this could pose a risk of
Instead, they
a theoretical study has failed to be field-tested.

More recently, Davies( 1993) described a computer
suggested that a more sensitive
index
the mean microfilarial load among adults aged 20
has become known
was
years
less effective control. This serves as an example of how
Community
model of onchocerciasis transmission( SIMON) based
to
widely
Further modeling, 12 to 14
initiated, indicated that because
on an original idea of D. F. Weidhaas, who had hoped it
very low CML' s successful control need only be

carried out for 15 years, not the 20 years originally
being unproved but had to wait until reliable field data

were available before it could be properly tested.
or more, which
Microfilarial Load( CML),
and
is
as the
now
used
evaluate control results.
would enable predictions to be made on the effects of
years after control was
different control measures. The model was continuously
of the
Remme
1990).
They
found
a good
Eventually, Davies ( 1993) was able to apply it to
observed
epidemiologicalandentomologicaldata collectedfrom
trends, and obtained evidence to support
a hyperendemic village in a forested area of Sierra
the hypothesis that the reproductive life span of the adult
Leone. The model was used to evaluate the effect ofboth
female worm was the main epidemiological determinant
vector
planned(
agreement
epidemiological
during
were
et al.
between
predicted
a vector control operation.
some
also
and
Nevertheless there
discrepancies between
predictions
and
and
chemotherapeutic
control
strategies,
separately and combined, and also the risk to an

uninfected population posed either by infectedSimulium
94
damnosum
house-spraying has, in many areas, not led to sustained
volvulus would
at six- month
be
malaria control. Because of these constraints and because
years could
house- spraying is useless against exophilic vectors,

there is continuing search for, and evaluation of,
if ivermectin
contrast,
intervals, it
in 18
eradicated
before 14
control
In
recrudescence.
delivered
to
O.
but abandoning
lead to
years
short distances. Even under optimum conditions, residual
the village.
entering
99
with
all adult worms of

years,
SIMON
percent effective vector control,
or people
that
predicted
were
up to 29
feasible
could take
to achieve eradication, although it might be

treatment after 18 years
stop
levels,
be 99
self-sustaining,
borne diseases, and the belief that there is no need for
both
more ecological data in order to undertake effective
percent as effective as vector con-
control, as argued for dengue, is the exception rather
drug
control,
disadvantage
trol, although a
ofnew control methods usually demands more ecological

information. The same is true for many other vector-
at reduced
vector and
could
alternative vector control measures. The implementation
residual
because the
longer be
worm population would no
Combined
DECEMBER, 1993
of
this approach is the
extended period of residual transmission and the slow
from the
elimination of worms
population.
The
infected immigrant flies were
than the rule.
Modeling has not generally led to the development
model
of better control strategies, and some models have
a greater
undoubtedly generated misleading ideas and
threat to the population than a small number of infected
conclusions. It is important to appreciate the limitations
entering the village. The plausibility

predictions is enhanced by the fact that
these
of models, but also to understand how, when fed with
relevant
from the
accurate and appropriate field-collected data, they may

be able to identify certain concepts relevant to control.
consequently be
For example, it must be determined which is the best
also showed that
of
people
data
epidemiological
were
collected
actually
village, and most parameters could
stage in a vector' s life- cycle on which to focus control,
estimated with considerable confidence.
the intervals at which control should be repeated, and
CONCLUSIONS
There
further
improving
Ae.
established between mathematical modelers and field
the only two important vectors of dengue
workers. If the isolation and barriers surrounding these
can
be few
generalizations on
ecological research on vectors
For
control strategies.
albopictus are
including
example,
Ae.
the need for
for
aegypti and
these species have very similar biologies,
two groups of people can be broken, then I believe that
the fact that throughout their range, larvae of
modeling may be able to assist in the planning of better
viruses, and
both
the critical vector or parasite densities below which the
disease dies out. In developing new models or expanding

the use of existing ones, it is essential that a more fruitful
dialogue and better understanding and respect are
in
occur
man- made and natural container- habitats.
vector control programs.
The consensus is that population levels, and the risk of

dengue outbreaks,
are
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DECEMBER, 1993
BORRELIA BURGDORFERI INFECTION IN IXODES SCAPULARIS

ACARI: IXODIDAE) IN KENT COUNTY MARYLAND
F. P. Amerasinghet and T. W. Scott2
ABSTRACT: Adult and immature stages of Ixodes scapularis Say( previously Ixodes dammini Spielman, Clifford,
Piesman, and Corwin) from a study area in Kent County, Maryland, were tested for infection with Borrelia
burgdorferi by immunofluorescence assay in 1991- 1992. Infection rates ranged from 27.0 percent to 45.2 percent
for different life stages, sexes, generations, and seasons of activity. The infection rates of engorged larvae attached
to white- footed mice( Peromyscus leucopus[ Rafinesque]) and the succeeding generation of questing nymphs were
similar, indicating that mice were the chief spirochete reservoir hosts in the area. Infection rates were significantly
higher in questing adults than in their nymphal progenitors, indicating that the tick population acquired a substantial
portion of infection during nymphal feeding. Estimates of the intensity of spirochete infection per tick( spirochete
burden) showed significant( P< 0.05) increases in spirochete numbers from larva to adult but not between nymphs
of
different
generations or adults of
different
seasons.
Thus, there was no evidence of decreased survival of
overwintering infected adult ticks. A significant, sex- related difference in spirochete burdens was seen in adults.
Studies from other areas of the I.scapularis geographic range would help to clarify seasonal and sex-related trends
in infection rates and spirochete burdens observed to date.
INTRODUCTION
Ixodes
Say ( reported
scapularis
synonym lxodes dammini

and
Corwin[ Oliver et
Lyme disease in
al.
investigated the rates and intensities of spirochete

infection in ticks of different life- stages, seasons, and
under
the junior
Spielman, Clifford, Piesman,
sexes at this same I.scapularis focus during 1991- 1992,

and our results are presented in this paper.
1993]) is the major vector of
the northeastern
United States( Lane et.
al 1991). Statewide surveys of adult ticks on whitetailed

deer( Odocoileus
fall deer- hunting
virginianus
season
reported as I.dammini)
have
Zimmerman)
shown
with
the
is well- established in the coastal
plains regions of eastern and southern
infected
during
that I. scapularis
Maryland, and is
the causative agent of Lyme disease, the
spirochete Borrelia
burgdorferi Johnson, Schmid, Hyde,
Steigerwalt,
and Brenner ( Amerasinghe et al. 1992,

1993). There is evidence that the distribution, abundance,
and rates of spirochete
may be
increasing
infection in deer- attached ticks
within the state(
Amerasinghe
et al.
1993).
life
Management Area( MWMA) in eastern Kent county,

Maryland( elevation
stages of the tick
exists on
the ecology of
in Maryland. A 1982- 1983
study at a focus of I.scapularis( reported as I.dammini)

infestation in the Millington Wildlife Management Area
MWMA) in Kent
white- footed mice(
County, Maryland,
10m above mean sea level; mean

12 C;
annual rainfall= 100 mm.

( National Oceanic and Atmospheric Administration
annual temperature=
[ NOAA]).
The abundant tree species in the study area
were white oak( Quercus alba), red oak( Quercus rubra),

sweet gum ( Liquidambar tulipfera), red maple ( Acer
rubrum),
pine (
However, little information

other
The study was carried out between June 1991 and

May 1992 in Zone 1 of the Millington Wildlife
tulip ( Liriodendron tulipfera), and loblolly
Pinus taeda).
The main understory plant was
coastal pepperbush( Clethra alnifolia).
Questing ticks were collected by dragging or

flagging on low vegetation using a 1 m2 white corduroy
cloth. Ticks were picked off the cloth at three minute
that
intervals during dragging/ flagging, which, for
Peromyscus leucopus[ Rafinesque])
convenience, are hereafter referred to as" drag" samples.
hosts of immature stages
These collections were done in the summer( June-July)

and fall ( November-December) of 1991, and spring
were the major small- mammal
showed
of this tick, and
that adults quested through the fall and
winter months
into the spring ( Haught 1983).
We
( March- May)
of
1992.
Immature 1. scapularis were
1Department of Zoology, University of Peradeniya, Peradeniya, SRI LANKA.

2Department
of
Entomology, University
of
Maryland, College Park, MD 20742, USA.
other
white- footed
from
collected
mice (
P. leucopus)
during
that were trapped
mammals
small
and
1 nymph on 2 of 3 masked shrews( Sorex cinereus Kerr);
the
4 larvae on 1 of 2 Norway rats ( Rattus norvegicus
in Sherman live traps
July- September) of 1991
summer(
intervals in trap lines within the study

The white- footed mouse has been previously
10
set out at
area.
in the study
the major natural
United States (
eastern
Haught 1983),
area(
and also
in Lane
reviewed
inhalant prior to tick removal,

capture after
Ticks
halothane
and replaced at the site of
recovery.
kept within humidified vials at4 C until
were
identification( based
on
Litwak 1989, Oliver

infection
spirochete
Midgut
is
1991).
et al.
trappedmammals were anesthetized with
smears of
Sonenshine 1979, Keirans
et al.
1993)
days
within seven
identified, live ticks
species that carried the greatest number of attached ticks
1991
and
and
1. 96+ 2. 10( 100 drags,
n=
196) in the spring
of 1992.
Borrelia burgdorferi infection was detected in 30
tested for
of
78( 38. 5%) engorged I.scapularis larvae and 2 of 6
direct immuno-
( 33. 3%) engorged nymphs removed from white- footed
fluorescence assay using fluorescein isothiocyanateconjugated polyclonal goat anti- B. burgdorferi
mice in the summer of 1991. The spirochete infection
the presence
of
B. burgdorferi
Questing I. scapularis nymphs averaged 0.82

per 3- min. drag sample( 100 drags; n= 82) in the
summer of 1991 and 0. 89 1. 34 per drag( 70 drags; n=
62) in the spring of 1992. Questing adults averaged 4.26
3. 06 per 3- min. drag( 80 drags; n= 341) in the fall of

1. 08
of collection.
were
Thus, the white- footed mouse
per host.
processing for
and
LeConte]).
was the most frequently caught small mammal within

Sherman live traps in the study area, as well as the
B. burgdorferi in the
reservoir of
Berkenhout]); 1 larva on a single pine vole( Microtus

pinetorum [
to be the major host of immature stages of I.
shown
scapularis
The
DECEMBER, 1993
100
by
rate of host- attached larvae did not differ significantly

from that of the succeeding nymphal stage questing in
KPL, Gaithersburg)( details in Amerasinghe
antibodies(
The intensity of B. burgdorferi infection in infected
the spring of 1992( TABLE 1)( X2= 0. 08, df= 1, P=

0.78). This suggested that white-footed mice were the
in three
major B. burgdorferi reservoir hosts infecting larvae in
et al.
1992).
ticks was estimated
fields
by counting
all spirochetes
320x
magnification
the study area although the rather unlikely possibility of
using the method described by Sharon et al.( 1992). For

consistency in estimation, all counts were made by one
other hosts carrying and infecting ticks at the same rates
microscope
worker (
three
The
1-PA).
fields
infection(
of view
under
burden)
spirochete
vs.
nymphs vs.
Two- way
nymphs).
1992:
comparisons of spirochete
burdens in
Since I. dammini has been

synonym of
name
is
used throughout the

studies
previous
published
et al.
text
Fall-
spirochete infection rate than their progenitor nymphs

questing in the
summer( X2=
4.01, df= 1, P= 0. 045),
indicating that a sizable proportion ofuninfected nymphs

acquired spirochetes during the nymphal feeding cycle.
spring
ANOVA) on
There was no significant difference in spirochete

infection rates of adult I. scapularis of the same
sex and season
generation that quested in different seasons ( fall 1991
adult ticks.
junior
vs. spring 1992: X2= 0.009, df= 1, P= 0.93). Spirochete

infection rates did not differ significantly between male
1993), the latter

referring to
of 1.
when
and
female ticks( fall 1991:
spring 1992:
the name
under
0.09) ( TABLE 1).
questing adults of 1991 had a significantly higher
relegated to a
I. scapularis( Oliver
2. 87, df= 1, P=
and
summer vs.
for
was used
X2=
infected
burdens( larvae
analysis of variance(
data
loge- transformed
adults,
Infection
for
ticks. Mann- Whitney U Tests were used for
nymphs,
excluded.
used
were
two- sample comparisons of spirochete
be
rates in different generations of questing nymphs were

not significantly different ( summer 1991 vs. spring
of
per tick.
Chi- square tests
statistical comparisons of proportions of

uninfected
intensity
was used as an estimate of the
Contingency
as white- footed mice cannot
average spirochete count of the
dammini.
X2=
X2=
0.23, df= 1, P= 0.63;
0. 004, df= 1, P= 0.95).
The estimated intensity of spirochete infection per

tick ( spirochete burden) was significantly higher in
RESULTS
questing nymphs in spring 1992 than in theirprogenitors,
the host-attached larvae of summer 1991( Mann-Whitney

A total
of
168 larval
was collected on
mean
of
SD= 3. 28
from 797
1991.
37
trap- nights
Data
on
29
nymphal
60( 61. 7%)
I. scapularis
U Test,
white- footed mice
4.54 immature ticks
per mouse)
in the
summer of
of collection
I. scapularis from
mammals were as
tailed shrews(
and
other
follows: 10 larvae
Blarina brevicaudata[
on
trapped
Say]);
of
short-
5. 13, df = 1,
P<
0. 001) ( TABLE 2).
Similarly, spirochete burdens were significantly higher

in fall 1991 questing adults than in their progenitors, the
summer
1991
nymphs ( z =
3. 11, df=
1, P = 0.002).
Infection intensities of different generations of nymphs
small
1 larva and
z =
summer 1991 and spring 1992) were not significantly

different(
z=
1. 51, df= 1, P= 0. 13)( TABLE 2).
DECEMBER, 1993
101
TABLE 1. Borrelia burgdorferi infection rates in questing adult and nymphal Ixodes scapularis.
Percent Infected( No. tested)
Total
Males
Females
Adults
Summer 1991
27. 0( 74)
Fall 1991
1992
Spring
TABLE 2. Mean
45. 2( 62)
39. 4( 66)
42.2( 128)
41. 8( 55)
42.9( 70)
42.4( 125)
number of spirochetes per tick
42.2( 64)
SE in adult and immature Ixodes scapularis.
Host- Attached
Questing
Questing
Questing
Total
Larvaea
Nymphsb
Males
Femalesc
Adults
75. 3 50. 8
100. 5 58. 0
87. 9 54.6
51. 9 29. 7
96. 2 53. 4
74. 1
Summer 1991
6. 57 6. 36
37. 1
31. 1
Fall 1991
1992
Spring
Nymphs
57. 1 48. 4
47. 8
aBased on 20 larvae.
bBased on 18 nymphs in 1991 and 20 in 1992.

Based on 10 specimens per sex in 1991 and 1992.
Note: Spirochete burden was significantly less in males than females ( fall 1991 and spring 1992 adults, 2- way
ANOVA, P= 0. 04).
Host-attached larvae of summer 1991 differed significantly from questing nymphs of
spring 1992( Mann- Whitney U test, P< 0. 001). All other comparisons within adult or nymphal cohorts were
not significantly different.
Statistical
comparisons( two- way
burden in
spirochete
questing in different seasons( fall 1991

showed
ANOVA)
of
the
adults of the same generation

vs.
spring 1992)
that season and sex- by- season interaction were
nonsignificant(
spirochete
P=
0.41
and
burdens in the two
0.54, respectively), but

sexes were
significantly
different( F= 4. 35; df= 1, 36; P= 0.04).
DISCUSSION
1991- 1992 in the MWMA. A previous investigation

done in the same zone of the MWMA showed a B.
burgdorferi infection rate of 35.6 percent( 54/ 157) in

guesting I.scapularis adults collected during November
1982 to March 1983. This rate did not differ significantly

from spirochete infection rates in adult ticks during the
fall of 1991 and spring of 1992 in our study( X2 tests, P
0.05). These findings suggest that our study was
carried out in an area of stable B. burgdorferi
transmission.
Borrelia burgdorferi infection rates and
burdens did not

generations
seasonally
change
spirochete
significantly between different
of nymphal
I.
scapularis, or
between
active adults of the same generation
during
Increasing rates of B. burgdorferi infection in I.
scapularis ticks from larva to adult in succeeding stages

of the same generation are to be expected since ticks
acquire
the spirochete from reservoir hosts
during
DECEMBER, 1993
102
of
2).
be
made in the period after a molt when the spirochete
extremely low ( Magnarelli et al. 1987, Piesman et al.

1986), and can be discounted as an important source of
density in ticks was much lower than after feeding to
larval infection in nature. Spirochete transmission from
increasing spirochete burdens from larva to adult also
successive
larval and
feeding cycles.
have been
Rates
shown
to
was seen
in Lane
in
infection
peak
role of adult hosts of I. scapularis( i.e., medium- sized
adult ticks.
mammals such as raccoons, skunks, and opossums,
this is important in
domestic animals, and large wild mammals, such as
in
deer, moose, bear, coyote, bobcat, etc.) as spirochete
in
rates
questionable whether
spirochete maintenance
of adult
correlate with the
of
seasonality
significance of this is uncertain, but it is possible that

hosts of adult ticks. Lane et al.( 1991) point out that the
et al.
their
I. scapularis in the
United States ( October- May) does
reservoirs needs more thorough evaluation.
We found that fall and spring questing adult ticks of

the same generation did not differ significantly in their
not
human Lyme disease,
spirochete infection rates and spirochete burdens contrary
is primarily a late spring and summer illness

peaking in July( Schmid et al 1985). Large and mediumwhich
mammals
like deer,
skunks,
raccoons,
to the results of Sharon et al.( 1992) in Wisconsin, which

showed significant decreases in both infection rates and
and
may be infected with B. burgdorferi( Anderson

1983; Bosler et al. 1984; Magnarelli et al. 1984,
spirochete burdens in overwintered adults. Sharon et al
opossums
1991), but this does

good reservoir
not
hosts.
and
spirochetes to
skunks
I.
of
lower
survive the winter. Such selection pressures were not
capable
are
scapularis
but
at
1992) suggested that substances important for spirochete
survival within ticks may be depleted during

overwintering, or that there may be a selective advantage
for the survival of uninfected or lightly infected ticks to
necessarily imply that they are

Fish and Daniels ( 1990) have
that raccoons
transmitting
1990), both
1985).
during
nymphs
disease epidemiology or in
nature. The guesting period
shown
et al. (
feeding
1991, Spielman
et al.
period results
et al.
Piesman
high spirochete inocula may be required to infect the
spirochete transmission
of more
sized
of
to humans
in
as
The infection
northeastern
in the study
1990). However, the trend of
post-repletion high- density phase. The epizootiological
play
in nature as well
However, it is
et al.
succeeding nymphs
in the maintenance cycle of B. burgdorferi
component of the cycle, since the
reviewed
Piesman
during the post-molt low spirochete density phase and
would
akey role
Our estimates for questing nymphs and adults were
repletion(
hosts ( primarily white- footed mice) to tick

be epidemiologically the most important
reservoir
larvae
nymphal
transmission
transovarial
e.,
manifested in Maryland. The explanation could lie in
contend
the less severe winter climate experienced by the more
that whitetailed deer, the majorhost of adult/. scapularis,
southerly situated Maryland tick population. Average
efficiency
than the accepted major reservoir
the white- footed mouse.
incompetent
are
decrease
Telford
1988)
for B. burgdorferi
reservoirs
spirochete transmission
Oliver
et al.(
host, i.
by
serving
1992) have
minimum temperatures in winter( December-February)
and
as
may
dead-
range from- 3 to- 6 C on the eastern shore of Maryland
that
experimentally inoculated whitetailed deer are capable

of transmitting B. burgdorferi to immature I. scapularis.
hosts.
end
et
al. (
asserted
PCR)
of
Administration). The milder environmental conditions
may impose less severe selective pressures on the

Maryland tick population, enabling both spirochete-
assays;
they did
not
viable spirochetes either
demonstrate the presence
in the deer
or
of a similar order of magnitude as
In
study
approximately
infected and uninfected ticks to survive with equal
those of
Sharon
were
success.
Sharon
et al
et al. (
1992) did not observe statistically
significant differences in spirochete loads related to tick
spirochete
methodology in Wisconsin.
burdens in ticks increased
eight- fold
between host- attached larvae
their data shows that in every instance males had a lower
who used the same
our
of
the ticks
Our estimates of the spirochete burden in ticks

1992)
9 to- 13 C in southwestern Wisconsin( based on data
published by the U.S. National Oceanic and Atmospheric
DNA by means of polymerase chain reaction
However, their conclusion was based on the detection

spirochete
where the MWMA is located), compared to values of
the point of acquisition) and the
succeeding questing
sex
in Wisconsin.
However, a close examination of
spirochete burden than females. Our results showed a
approximately two- fold between questing
significant sex difference in spirochete burdens ( P =
the succeeding questing adults. Spirochete
0.04 level, with nonsignificant interaction effect) in
in host- attached larvae may have

the true spirochete burden
both fall and spring questing adults of the same
during feeding since the ticks, though engorged,
method of spirochete estimation was adequate for
detached naturally from the host at the time of

testing. However, it is clear that the spirochete burden
increased in successive life stages of the tick( TABLE
detecting large differences in spirochete burdens;

however, it may not be sufficiently reliable to make firm
nymphs, and
nymphs and
numbers recorded
been
an underestimate of
acquired
had
not
generation. As Sharon et al.( 1992) have pointed out, the
conclusions
based
on
the narrower differences in
spirochete numbers observed
Studies from
resolving the
between the tick
sexes.
other northern and southern areas of the
geographic
scapularis
103
DECEMBER, 1993
issues
range
I.
be helpful in
would
of lowered survival of overwintering
Keirans, J. E. and T. R. Litwak. 1989. Pictorial keys to
the adults of hard ticks, Family Ixodidae( Ixodida:

Ixodoidea), east of the Mississippi River. J. Med.
Ent. 26: 435- 448.
infected ticks and the differential spirochete burdens in

male and
et al.(
female ticks that have been
1992)
by Sharon
raised
Lane, R. S., J. Piesman,
and
W. Burgdorfer.
1991.
Lyme borreliosis: relation of its causative agent to
and the present work.
its vectors and hosts in North America and Europe.

Ann. Rev Ent. 36: 587- 609.
Acknowledgments
We
Brian Colhoff, Leslie Lorenz,
thank
Breisch, Lynn Cooper, John Putnam,

Mallampalli for field
1991.
Funding
and
during the
provided by the
assistance
was
Nancy
Varuni
W. A. Chappell.
1984.
Parasitism by Ixodes
summer of
dammini ( Acari: Ixodidae) and antibodies to
Maryland
spirochetes in mammals at Lyme disease foci in
This is Scientific
Agricultural Experiment Station.
Magnarelli, L. A., J. F. Anderson, W. Burgdorfer, and
Connecticut. J. Med. Ent. 21: 52- 57.
Article No. A6477 ( Contribution No. 8684) of the

Magnarelli, L. A., J. F. Anderson, and D. Fish. 1987.
Maryland Agricultural Experiment Station.
Transovarial transmission of Borrelia burgdorferi

in Ixodes dammini( Acari: Ixodidae). J. Infect. Dis.
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156: 234- 236.

Amerasinghe, F. P., N. L. Breisch, A. F. Azad, W. F.
Gimpel, M. Greco, K. Neidhardt, B. Pagac, J.
Piesman, J. Sandt, T. W. Scott,
1992.
Distribution,
and
Sweeney.
Lyme disease
infection in Ixodes dammini ( Acari:
spirochete
Ixodidae)
density,
K.
and
on
whitetailed
deer in Maryland.
Magnarelli, L. A., J. H. Oliver, H. J. Hutcheson, and J.

F. Anderson.
1991.
562- 568.
J.
Oliver, J. H., D Stallknecht, F. W. Chandler, A. M.
Med. Ent. 23: 54- 61.
James, B. S. McGuire,
Amerasinghe, F. P., N. L. Breisch, K. Neidhardt, B.

Pagac,
and
T. W. Scott. 1993.
Increasing density
Borrelia burgdorferi infection
and
Antibodies to Borrelia
burgdorferi in deer and raccoons. J. Wildl. Dis. 27:
of
Ixodes
dammini( Acari: Ixodidae) in Maryland. J. Med.
and
E. Howerth.
1992.
Detection of Borrelia burgdorferi in laboratoryreared Ixodes dammini ( Acari: Ixodidae) fed on

experimentally inoculated
whitetailed
deer.
J.
Med. Ent. 29: 980- 984.
Ent.( in press).
Anderson, J. F., L. A. Magnarelli, W. Burgdorfer,
and
A. G. Barbour. 1983. Spirochetes in Ixodes dammini

and
mammals
Med.
Hyg.
from Connecticut.
Am. J.
Trop.
and
J. L. Benach. 1984. Prevalence
of
the Lyme disease spirochete in populations of whitetailed deer and white- footed mice.
Yale J. Biol.
and
T. J. Daniels. 1990. The
southern
J.
Piesman, J., J. G. Donahue, T. N. Mather, and A.
Spielman.
1986. Transovarially acquired Lyme
disease spirochetes( Borrelia burgdorferi) in fieldcollected larval Ixodes dammini( Acari: Ixodidae).
role of medium-
Piesman, J., J. R. Oliver, and R. J. Sinsky. 1990. Growth
burgdorferi
kinetics of the Lyme disease spirochete( Borrelia
sized mammals as reservoirs of Borrelia
in
I. dammini ( Acari: Ixodidae).
J. Med. Ent. 23: 219.
Med. 57: 651- 659.

Fish, D.
scapularis and
Med. Ent. 30: 54- 63.
32: 818- 824.
Bosler, E. M., B. G. Ormiston, J. L. Coleman, J. P.

Hanrahan,
Oliver, J. H., M. R. Owsley, H. J. Hutcheson, A. M.

James, C. Chen, W. S. Irby, E. M. Dotson, and D.
K. McLain. 1993. Conspecificity of the tick Ixodes
New York. J. Wildl. Dis. 26: 339-
345.
burgdorferi) in vector ticks( Ixodes dammini). Am.
J. Trop. Med. Hyg. 42: 352- 357.
Haught, S. B. 1983. Ixodes dammini( Acari: Ixodidae)
Schmid, G. P., R. Horsley, A. C. Steere, J. P. Hanrahan,
in Maryland: the potential for Lyme disease. M. Sc.
J. P. Davis, G. S. Bowen, M. T. Osterhohn, J. S.
Thesis,
University
of
Maryland.
vi+
49 pp.
Wiesfeld, A. W. Hightower,
and
C. V. Broome.
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1985.
Surveillance
United States,
1982.
Lyme disease
in
the
Sciences, Agricultural Experiment Station, Virginia.
J. Infect. Dis. 151: 1144-
Polytechnic Institute and State University,
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Sharon, M. D., W. A.
DECEMBER, 1993
Blacksburg, 44 pp.
Rowley, M. G. Novak, and K. B.
Spielman, A.,M. L. Wilson, J. F. Levine, and J. Piesman.
Platt. 1992. Rates of Borrelia burgdorferi infection
1985. Ecology of Ixodes dammini-borne human
in Ixodesdammini( Acari: Ixodidae) in southwestern
babesiosis and Lyme disease spirochete. Ann. Rev.
Wisconsin. J. Med. Ent. 29: 314- 317.
Ent. 30: 439- 460.
Sonenshine, D. E. 1979. The insects
of Virginia: No 13.
Ticks of Virginia( Acari: Metastigmata). Research
Division Bulletin No. 139.
Entomology, College
of
Department
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and
of
Life
Telford, S. R. III, T. N. Mather, S. I. Moore, M. L.

Wilson, and A. Spielman. 1988. Incompetence of
deer as reservoirs of the Lyme disease spirochete.
Am. J.
Trop. Med. Hyg.
39: 105- 109.
DECEMBER, 1993
SIZE OF EMERGING AND HOST-SEEKING AEDES AEGYPTI

AND THE RELATIONSHIP TO CONTAINERS AND BLOOD-
FEEDING SUCCESS IN TRINIDAD, WEST INDIES.
Dave D. Chadeel
ABSTRACT: The wing lengths ofAedes aegypti females collected as pupae from cans, basins, buckets, and drums
averaged 2.42 mm, 2.61 mm, 2. 61 mm, and 2. 65 mm, respectively. Females with the largest wing lengths were
collected from drums( F=93. 62, P< 0.001). The wing lengths of adults emerging from drums ( P< 0.001), buckets
P< 0.01), and basins( P< 0. 01) were significantly larger than those
emerging from cans( P= 0. 21). The wing lengths
ofparous host- seeking females collected indoors were larger( 2. 59 mm) than parous females collected outdoors( 2. 53
mm) but this difference was not significant. In contrast, the nulliparous females collected outdoors( 2.49 mm) were
significantly( P< 0.01) larger than the nullipars collected indoors( 2. 36 mm). However, both nulliparous and parous
host-seeking females collected outdoors were similar in size when compared( 2.49 vs. 2.53 mm). The data suggested
that small Ae. aegypti females exhibit reduced blood- feeding success indoors and possibly reduced survival when
compared to large adults.
INTRODUCTION
was studied in St. Joseph( 10 38' N; 61 25' W), Trinidad,
W.I. Details of the study area, vegetation, topography,

Container
size
due to
breeding
mosquitoes
factors
numerous
such
vary markedly in
meteorology, and Ae. aegypti population have been

described by Chadee and Corbet ( 1987). Collections
as
overcrowding
Mulla 1979), temperature ( Brust 1967), mutual
interference ( Terzain
and insufficient
the
larval
However, the
be
amount of variation
related to the
in
type of larval habitat

or
habitats may display more variance in adult

than do species using more permanent habitat
ephemeral
body size
sites. Consequently, individual size is an important part
of the life- history strategy of a species and is often
related to survivorship and fecundity ( Begon et al.
1986). Nasci( 1986) reported great variations in wing
length among emerging Aedes aegypti males and
females and concluded that small females may exhibit
reduced blood- feeding success and most
likely reduced
survival when compared with
purpose of this
large
1993) from a total of 1, 350 houses. Aedes aegypti pupae
during
Fish 1985). Thus, mosquitoes using small containers
The
were made daily for 28 weeks ( July 1992- February
Stahler 1949, Mulla 1979)
food( Shannon and Putnam 1934)
stage.
size appears to
and
adults.
was to examine the
study
wing
length ofAe. aegypti adults emerging from four different
were collected from drums( capacity 208 litres), buckets

capacity 22. 10 litres), basins( capacity 9. 10 litres), and
cans( capacity 800 mis) per occasion. These containers
were all located outdoors. In the laboratory the pupae

were held at 28 1 C in rearing dishes and placed into
separate emergence cages labeled according to the
containers from which they were collected. Laboratory
conditions and rearing methods used were in accordance
to those described by Chadee ( 1992).
Twenty- four
hours after adult emergence, the wings were removed
and placed on a slide in a small drop of dilute saline

solution. The distance from the axillary incision to the
apical margin excluding the fringe scales on each
mounted wing was measured as described by Nasci
( 1986) using a binocular microscope with an eyepiece
graticule and slide micrometer.
determine differences, if any,

in their wing length with that of host- seeking females
captured in the field.
Host- seeking females were collected from human

bait using aspirators, flashlights, and hand net between
1600 and 1800 hours, the evening peak in the Ae.
aegypti biting cycle ( Chadee 1988). Collections were
made at four sites. At each site two collectors were
sizes of containers and
to
stationed outside in the yard and two indoors in the

The wing
collected pupae and
Ae.
emerging from field
living room. All mosquitoes caught on exposed lower
host seeking towards human bait
legs and ankles of the collectors were transferred into
size of
aegypti
llnsect Vector Control Division, Ministry
of
Health, 3 Queen Street, St. Joseph, Trinidad, WEST INDIES.
jars lined
with plaster of paris and were
collector and
In the
kept separate
The wing lengths of parous and nulliparous Ae.
females
by
parous
x magnification,
sites are compared in TABLE 2. The parousAe. aegypti

females captured indoors averaged 2. 56 mm and were
Ae.
larger than the parous outdoor females ( average 2.53
were examined and scored as nulliparous
mm), but this difference is not significant. The nulli-
The
Detinova' s
or absence
presence
1962). Mosquito
for
aegypti collected host seeking at indoor and outdoor
wings were prepared and
and counted.
40
their lengths measured as described above while all
method,
to
Data
the
parous
according
Detinova
collections were analyzed
were analyzed
using
collected
outdoors (
P> 0. 01)
were
In addition, parous females collected indoors were
separately
( indoor or
an analysis of var-
females
significantly larger than the nullipars collected indoors.
of tracheolar skeins (
each container type and collection site
outdoc:).
when compared.
laboratory the mosquitoes were anesthetized,
identified,
aegypti
by
location.
examined under a microscope at
or
DECEMBER, 1993
106
larger than nullipars collected both indoors and outdoors

(
F= 34. 8; P> 0.01)( TABLE 2).
iance( Sokal and Rohlf 1980) to determine differences

in mosquito wing lengths from different size
DISCUSSION
containers
and parity determinations.

The present study demonstrated that wing length of
Ae. aegypti varied significantly in size depending on
RESULTS
their container habitats. Nasci( 1986), however, found
Wing
length
measurements of
Ae.
in
a large range of body size among Ae. aegypti in a tire

dump in the U.S. A. A possible explanation may be that
No
tires were of different sizes and their position on the
wing
detected
ground or in a tire dump may have influenced the
lengths
for each pre- adult was different in each tire which may
aegypti adults
that emerged from drums, buckets, basins, and cans
St. Joseph, Trinidad

significant
lengths
are presented
in TABLE 1.
differences between left
of male and
when compared.
were collected
female Ae.
Females
with
and
right
aegypti were
the largest wing
from drums ( F- 93. 62, P< 0. 001).
The
wing length of adults emerging from drums( P< 0. 001),

buckets( P> 0. 01), and basins( P> 0.01) were significantly
larger than those emerging from cans. However, there
difference in wing lengths among

females emerging from buckets and basins ( P= 0. 31)
was no significant
TABLE 1.
amount of water accumulated. That is, food and space
affect the size of adult mosquitoes.
In contrast, size
variation in Ae. aegypti can be genetically influenced

( Greenough et al. 1971).
Christophers( 1960) showed that wing length was
directly related to body size in Ae. aegypti mosquitoes.

Field studies have shown that adult mosquitoes with
Wing length of emerging Aedes aegypti collected from drums, buckets, basins, and
cans in the field.
Wing length( mm)

Container
Sex
Sample
Average
Emerging
Emerging
247
2. 05+ 0. 05
1. 96- 2. 15
269
2. 65+ 0. 09
2.43- 2. 88
Emerging
Emerging
353
2. 03+ 0. 01
1. 74- 2. 31
335
2. 61+ 0. 01
2. 15- 2. 99
Emerging
Emerging
273
2. 02+ 0. 04
1. 84- 2. 12
267
2. 61+ 0. 07
2. 30- 2. 78
193
1. 93+ 0. 06
1. 74- 1. 74
208
2. 42+ 0. 09
1. 96-2. 78
Drums
Male
Drums
Female
Buckets
Male
Buckets
Female
Basins
Male
Basins
Female
Cans
Male
Cans
Female
Emerging
Emerging
Range
DECEMBER, 1993
TABLE 2.
107
Percentage of the Aedes aegypti population that was parous and
mean wing length of the parous and nulliparous.

Percent
Sample
Host- seeking
Mean wing length( mm)
n)
Parous
Parous
Nulliparous
100)
28
2. 59+ 0.06
2. 36+ 0. 05
105)
21
2. 53+ 0. 07
2. 49+ 0. 05
indoors)
Host- seeking
outdoors)
relatively larger bodies ( correlated with larger wing

lengths) exhibit higher fitness qualities than do small-
bodied mosquitoes may contribute more to maintaining
bodied females. Larger Aedes triseriatus(
a mosquito population with large average wing length

may have a higher vectorial capacity than a population
are
frequently
more
1981), larger Aedes
Milby
fecundity(
sierriensis (
have higher survivorship rates( Nasci, 1986),

study largerAe.
females
Haramis 1983), larger
parous (
Culex tarsalis Coquillett have higher

and
Say)
Bock
Ludlow)
and
in this
and
parity
with a small average wing length( Nasci 1986).

It is clear from these results that small ( cans),
medium (
buckets), and large ( drums) containers all
to enter houses
produce Ae. aegypti capable of becoming parous
the present study, variations in wing length
for mosquito control whether by chemical, biological,
aegypti were more
likely
to blood feed.
During
and amplifying arboviruses than small individuals, and
of
individuals. Therefore, these containers must be targeted

Ae.
aegypti were
found,
and these were
or source reduction methodologies.
related to blood- feeding success. In addition, the wide

range of wing lengths found among Ae. aegypti
collected may be related to the size ofcontainers,
ACKNOWLEDGMENTS
quality
and quantity of food resources available( Shannon and

Putnam 1934),
and the
each container(
Mulla 1979, Terzan
in
Special thanks are due to Messrs A. Mohommed
Stahler 1949).
and Robin C. Persad for assistance in the laboratory and
density of pre- adults present

and
It is noteworthy that Ae. aegypti of all wing lengths

were collected at indoor and outdoor sites( TABLE 2).
At indoor
sites,
however, individuals
were
larger in
field.
REFERENCES
size
and more likely to be parous, indicating that they

obtained
successfully
suggested
distances, are
blood
Nasci ( 1986)
meals.
individuals may fly for
that small
short
not as persistent, or exhaust their reserves
Begon, M., J. L. Harper, and C. R. Townsend. 1986.
Ecology:
Individuals,
Populations
and
Communities. Blackwell Sc. Publ. Oxford.
sooner than larger Ae. aegypti females. In addition,

Klowden
et al.(
1988) demonstrated that large
aegypti that were
fed
on an optimal
diet
adult Ae.
were more
likely to engage in host seeking than small females from
Bock, M. E. and M. N. Milby. 1981. Seasonal variation

of wing length and egg raft size in Culex tarsalis.
Proc. Calif. Mosq. Vector Contr. Assoc. 49: 64-66.
poorly fed larvae. Data from the present study showed

that
female Ae.
smaller
aegypti collected as pupae
in
cans
had
wing lengths than parous host- seeking females,
suggesting that mortality prior to blood feeding was

higher in the smaller adults than in the larger individuals.
Brust, R. A. 1967. Weight and development time of

different stadia of mosquitoes reared at various
constant
temperatures.
Can. Entomol. 99: 986-
993.
However, in drums, buckets, and basins large females

were produced and were similar
host- seeking females
collected.
in size to the
These
parous
results suggest
that large containers produce females that are able to

enter
houses
and contact more
hosts. Therefore large-
Chadee, D.D. 1988. Landing periodicity ofthemosquito,

Aedes
aegypti ( L.)
in Trinidad in relation to the
timing of insecticides space spraying. Med. Vet.

Entomol. 2: 189- 192.
DECEMBER, 1993
108
1992. The life
Chadee, D. D.
of
table characteristics
laboratory population of Aedes

J. FloridaMosq. Contr. Assoc. 63: 68-
selected
aegypti(
L.).
Haramis, L. D.
1983.
Increased adult size correlated
with parity in Aedes triserratus. Mosq. News 43:

77- 79.
72.
Klowden, M. J., J. L. Blackmer, and G. M. Chambers.

Chadee, D. D.
incidence
field
and
and
P. S. Corbel.
diel
1987.
Seasonal
1988. Effects of larval nutrition on the host-seeking
in the
behavior ofadultAedes aegypti mosquitoes. J. Am.
patterns of oviposition
of the mosquito,
Aedes
aegypti(
L.)( Diptera:
Mosq. Contr. Assoc. 4: 73- 75.
Culicidae) in Trinidad, West Indies: a preliminary

study. Ann.
Trop. Med.
Christophers, S. R.
Parasitol. 81: 151- 161.
1960. Aedes
yellow fever mosquito.
aegypti (
L.). The
Mulla. M. S. 1979. Chemical ecology of mosquitoes

auto and transspecific regulating chemicals in nature.
Proc. Calif. Mosq. Contr. Assoc. 47: 65- 68.
Cambridge Univ. Press.

Nasci, R. S.
London, 737 pp.

Detinova, T. S. 1962. Age grouping methods in Diptera
of medical importance. W.H. O. Monogr. Ser. 47,
1986. The size of emerging and host

seeking Aedes aegypti and the relation of size to
blood- feeding success in the field. J. Am. Mosq.
Contr. Assoc. 2: 61- 62.
216 pp.
Fish, D. 1985. An analysis of adult size variation within
populations. Ecology of
Pp. 419-429 in Proceedings of a
Workshop( L. P. Lounibos, J. Rey, and J. H. Frank,
natural
mosquito
Shannon, R. C. and P. Putnam. 1934. The biology of

Stegomyia under laboratory conditions.
Proc.
Entomol. Soc. Wash. 36: 185- 216.
Mosquitoes.
eds.).
Florida Medical Entomol. Lab. Vero Beach,
Florida, 579 pp.
of
and
P. Barbosa. 1971.
survival, weights and pupation rates
four Aedes
aegypti ( L.). strains reared with a
standard technique.
and
F. J. Rohlf.
1980.
Biometry. The
principles and practice of statistics in biological
research. 2nd. Freeman, New York, 859 pp.
Greenough, N. C., T. M. Peters,

Comparative
Sokal, R. R.
J. Med. Entomol. 8: 502- 503.
Terzian, L. A. and N. Stahler. 1949. The effects of larval

population
density
on
some
laboratory
characteristics ofAnopheles quadrimaculatus Say.

J. Parasitol. 35: 487- 495.
DECEMBER, 1993
DAILY SURVIVORSHIP AND LIFE SPAN OF THE MOSQUITO
ERETMAPODITES QUINQUEVITTATUS THEOBALD (DIPTERA: CULICIDAE)

UNDER LABORATORY CONDITIONS COMPARED TO
AEDES AEGYPTI, AEDES ALBOPICTUS, AND AEDES BAHAMENSIS
A. M. Helleck1, W. K. Hartberg1, and D. Vodopichl
This laboratory study analyzes average daily survivorship and adult life span of Eretmapodites
Abstract:
quinquevittatus, Aedes aegypti, Aedes albopictus, and Aedes bahamensis. Experimental populations of 25 adult
males and 25 adult females were observed, as well as experimental pair groups of 25 pair cages containing one male/
one female each. Life spans of blood fed and non-blood fed populations of Er. quinquevittatus were compared to
those ofAe. aegypti, Ae. albopictus, andAe. bahamensis. Average daily survivorship for males of Er.quinquevittatus
was
significantly
that of all the Aedes
greater than
species
regardless
of
feeding
treatment.
Average daily
survivorship for females of Er. quinquevittatus was significantly greater than that of Ae. aegypti in the non-blood
fed treatment. Non-blood fed average daily survivorship for Ae. bahamensis was significantly greater than that of
the other two Aedes species, and not significantly different than that of non-blood fed females ofEr. quinquevittatus.
Survivorship forAe. aegypti was greater when males and females were blood fed, which conflicts with earlier reports.
Unexpectedly, survivorship and longevity of Er. quinquevittatus males was significantly greater than that of other
commonly studied mosquitoes.
Hylton 1969, Hartberg and Gerberg 1971) and because
INTRODUCTION
members of this genus can transmit pathogens( Bauer

Our study examined adult survivorship of
Eretmapodites quinquevittatus Theobald ( Diptera:
Culicidae),
a species native to subsahara
Madagascar that may transmit

Previous
and Gerberg 1971).
is easily
quinquevittatus
an excellentorganism
reared
Africa
studies
in the
and
Hartberg
arboviruses (
Er.
show that
laboratory and is
and Gerberg
forresearch( Hartberg
1971).
This study
for Er.
compared average
when
quinquevittatus
experimental populations or
analyzed
the effect of blood
survivorship for these
daily
adults
individual
feeding
species.
survivorship
held in
were
We
pairs.
on average
We
also
daily
compared adult
survivorship of Er. quinquevittatus to that of three

species, Aedes aegypti ( Liles and Delong 1960,
Christophers 1960, Gillett 1971),
Hylton 1969, Gubler 1970, Gao
1985,
and
Hawley
1988),
Weyer 1965).
because Aedes
and
Edwards 1941,
and
These
Hartberg
et al.
1984, Liu
are related genera
history
of
1Department
of
Baylor
Although this species is not endemic to
introduction with international trade and travel. Basic
data on life span, developmental needs, fecundity, etc.

are best gathered initially in the laboratory where they
may be essential to later field research. Furthermore,

preliminary studies indicate that males and females of
Er. quinquevittatus may be long- lived ( Hartberg and
Gerberg 1971), and longevity can be an important factor
in vector significance.

Cultures
were maintained at
25 C, 80 5 percent
RH, 14 h light, and 10 h dark. Populations ( 25 adult

males and 25 adult females) were held in cylindrical,
gallon cages( 4329. 5 ml) and pairs( one adult male- one
the
cages ( 690. 1 ml) covered with nylon netting. Mason
completed
jars( half-pint, 279. 3 ml) or beakers( 250 ml) with 150-
studies on
Eretmapodites have been
Biology,
1968).
North America there is always the potential for
adult female) were each held in cylindrical, pint-size
Faircloth 1983).
Our report is important because few

life
et al.
comparisons are relevant
and
1965, Gilotra and Shah 1967, Brottes et al. 1969, Serie

et al.
Aedes albopictus
Aedes bahamensis( Spielman
Eretmapodites
1928, Smithbum et al. 1948, Macnamara 1953, McIntosh

et al. 1961, Worth et al. 1961, Ardoin and Simpson
University,
Waco, Texas 76798, USA.
110
200
tap water were placed in
ml of
the population cages
DECEMBER, 1993
RESULTS
for oviposition. Small glass vials( 42.5 ml) with 40 ml

tap
of
inside the
water were placed
oviposition containers were

paper
towel for
To
populations
from the
were obtained
at
Blood Feeding and Population Density Effects on

Eretmapodites quinquevittatus
white-
eggs.
receiving
establish
All
pair cages.
lined with bleached,
Average daily survivorship and adult life span of

Er. quinquevittatus were calculated from observations
and pair groups, eggs
following
colonies maintained
of two density treatments: 16 experimental populations
the Medical Entomology Research Laboratory, Baylor
University: EQ- MIXED

ROCK
strain of
strain ofAe. aegypti,
the BU strain of Ae.
and
Survivorship for blood fed and non-blood fed
HC strain ofAe. albopictus,
females of Er. quinquevittatus did not differ( P>0. 05).
bahamensis. The
resulting pupae were
cylindrical,
pint- size
eggs were
Survivorship was also not different for females held in
sexed and assigned to
population ( large) or pair ( small) cages ( P> 0.05).
emergence
These
and two experimental pair groups( TABLE 1).
quinquevittatus,
cultured and
emerged(>_10/ cup).
Er.
adults
Survivorship for males held with blood fed and non-
adults were then assigned to
blood fed females was not different ( P> 0. 05).
cups
where
Days lived by
population and pair cages.
were used to calculate average
Survivorship was not different for males held in
these adults
daily survivorship
population or pair cages( P> 0.05).
and
mean life span.

The colony adults were provided with beakers lined
with bleached paper
towel for oviposition.
in the same beakers in
instar larvae
30
26
with>
cm x
250
ml
were
fed
30 eggs
6 cm),
eggs were reared
cm),
beakers,
of
slurry
half-pint
or
for the
blood to
in large pans
mason
data.
pint- size
Female survivorship was significantly different

among the four
jars. Larvae
Tetramin fish food. Pupae
top
adults
adult
and
quinquevittatus was significantly greater than the
survivorship of non-blood fed females of Ae. aegypti
PLSD P< 0.05). Survivorship ofnon- blood fed females

of Ae. aegypti was also significantly less than
survivorship of non- blood fed females ofAe. albopictus
Non- blood fed
and
production
anautogenous
meal
was
Twelve hours
after
production subsided another
blood
subsided.
egg
given.
blood
Subsequently,
every four to five days.

Dates of emergence and death
meals
than that of Ae. aegypti non-blood fed females ( PLSD

There were no significant differences in
P< 0.05).
survivorship when females were blood fed.
Male survivorship was significantly different among
were
provided
foreach adult.
for
the
were
group( 25
pair cups with
each) and recorded as the proportion of
also calculated
for males
each
and
day.
Mean life
Survivorship for Ae. bahamensis males held with nonblood fed females was significantly greater than the
survivorship of Ae. aegypti and Ae. albopictus males
span was
females of each population
and pair
group.
Effects of
contained with non- blood fed females( PLSD P< 0.05).

container
density
on adult
2- way ANOVA
with
transformed average
size,
life
blood
span were
replication
daily
feeding,
using
survivorships.
determine
Survivorship for Ae. aegypti males held with blood fed
and
determined
protected least significant differences tests(
to
Survivorship for
than for all three Aedes species whether held with

females blood fed or non-blood fed ( PLSD P< 0.05).
surviving
by
females was significantly greater than survivorship of
arcsine
Ae. aegypti males held with non- blood fed females
Fischer
PLSD)
were
which comparisons
among
differed.
Alpha and power for the
significantly
statistical tests were 0. 05 and 0.80,
respectively.
species
P< 0. 001).
Averagedaily survivorship was calculated
1 female
performed
species compared(
males of Er. quinquevittatus was significantly greater
the population
mosquito
four
documented
each population and pair
male/
Ae. bahamensis( PLSD P< 0. 05).
Survivorship of
blood fed Ae. aegypti females was significantly greater
only sugar. Blood meals

forbloodfedpopulations orpairs began when autogenous
populations or pairs received
egg
P< 0. 001).
were
anesthetized mice
females.
compared (
emergence cups
the cage provided
of
species
Survivorship of non- blood fed females of Er.
10/ cup).
cubes placed on
carbohydrate
to that ofAe. aegypti,Ae. albopictus, andAe. bahamensis
( TABLE 1). All comparisons weremade withpopulation
progeny from papers with

in small pans( 26 cm x 16 cm x 6
where adults emerged(?
provided
were reared
Progeny
while
transferred to cylindrical,
Sugar
to Nalgene pans where
continued to the pupal stage.
from egg papers

x
they were oviposited. First
were transferred
development
32 cm
which
Survivorship of Eretmapodites quinquevittatus and

Aedes spp. in Population Cages
Survivorship of Er. quinquevittatus was compared
Eggs hatched
PLSDP<0.05). Survivorship forAe. bahamensismales

held with non- blood fed females was significantly greater
than survivorship of Ae. bahamensis males held with
blood fed females( PLSD P< 0.05). Survivorship for Er.

quinquevittatus and
Ae.
albopictus, within species,
did
DECEMBER, 1993
TABLE 1.
Average
daily
survivorship(%)
111
and adult life span ( days) for Eretmapodites quinquevittatus, Aedes
aegypti, Aedes albopictus, and Aedes bahamensis held in populations( 25 males/ 25 females each) and
pairs (
male/
1 female
each- 25
pairs
in
group) in the
laboratory.
Standard deviations are in
parentheses.
Avg. Daily
Male
Eretmapodites
quinquevittatus
Populations Blood Fed

N= 10)
Populations Non- Blood Fed

N= 6)
Pairs Blood Fed
Pairs Non- Blood Fed
Aedes
aegypti
Sur(%)
Female
Mean Life Span

Male
Female
96.4
97. 8
37. 6( 31. 8)
96.6
97. 4
57. 4( 38. 1)
77. 0(40.4)
96. 3
95. 1
38. 5( 33. 9)
25. 9( 24.8)
96. 6
96. 3
30.6(37.0)
38. 7( 31. 8)
96.9
97. 7
60. 2( 24. 7)
57. 5( 35. 3)
94.9
96. 7
35. 3( 26. 7)
47. 0( 32.2)
95. 5
95. 7
48. 0(26. 3)
41. 4( 21. 2)
96.4
90. 5
40. 2( 34. 3)
12. 8( 8. 3)
95. 8
94.4
42. 8( 28.9)
22.2( 17.4)
96. 1
91. 9
24. 6( 26. 1)
14.7( 7. 5)
96. 8
96. 2
56. 7( 35. 1)
43. 0( 31. 1)
96. 2
85. 7
54. 0(27. 4)
61. 0( 10.5)
96. 5
95. 8
50. 0( 35. 6)
44.3( 25. 3)
96. 2
97. 0
47. 2( 24. 8)
49. 2( 33. 1)
96. 1
96. 0
42. 9( 28. 8)
59. 7( 30.9)
95. 7
95. 7
42.9( 28.4)
55. 9( 22. 5)
96. 6
97. 5
67. 2( 34. 7)
47. 5( 45. 2)
96.9
97. 5
51. 6( 40. 9)
53. 7( 40.2)
96. 8
96.9
62. 8( 37.9)
75. 7( 30.8)
97. 3
96.6
77. 6( 38. 1)
71. 6( 30.2)
83. 4
92. 1
8. 3( 3. 2)
19. 8( 9. 5)
85. 0
97. 5
8. 4( 4.2)
57. 2( 40.2)
91. 2
96.9
9. 7( 7. 0)
60.9( 39.4)
53. 1( 49. 7)
82. 6
86. 6
8. 1( 3. 4)
20.3( 4. 9)
77.6
88. 3
9. 0( 2.8)
17.4( 6.9)
77. 7
87. 0
9. 4( 2.9)
16.9( 5. 9)
albopictus
83. 8
97. 4
11. 9( 4. 8)
57.6(42. 7)
80. 9
96.7
11. 2( 4.6)
45.0(35. 8)
91. 0
96.8
15. 1( 6.6)
46.2( 41. 1)
90.2
96. 1
14. 8( 9. 3)
49.2( 26.7)
85. 3
95. 7
10. 2( 4. 9)
43. 7( 32. 5)
84. 1
95. 6
11. 2( 26. 7)
41. 0( 33. 7)
Aedes bahamensis
83. 8
97. 3
13. 7( 5. 3)
63. 6( 44.9)
90.0
96.9
20.4( 8. 4)
65. 3( 33. 1)
94. 1
97. 1
33. 0( 18. 6)
44.0(38. 2)
92. 3
96.2
19.8( 12. 3)
52.0(26.0)
95. 5
96.2
29. 4( 20.6)
61. 8( 24. 5)
92.6
96.2
20. 3( 11. 9)
61. 7( 24. 5)
Aedes
112
not
differ due to
feeding
DECEMBER, 1993
Edwards, F. W.
treatment.
1941.
Mosquitoes of the Ethiopian
Region III: Culicine adults and pupae. London:

DISCUSSION
Average
daily
quinquevittatus
the
other
did
survivorship for females of Er.

differ from the survivorship of
not
Aedes females,
quinquevittatus survived
only do the
British Museum( Nat. Hist.),
while
P. Zhao,
and
N. H. Cao.
1984.
Studies on the
Er.
males.
Not
longevity of adult Aedes ( S) albopictus ( Skuse):

the longevity of caged females under laboratory
longer
conditions. Acta Entomol. Sinica 27: 182- 188 ( in
males of Er. quinquevittatus survive
Aedes, but they also survive as long as

females of Er. quinquevittatus andAedes. Furthermore,
than males of
the ingestion of blood

a role
Gao, J. Z., Z. Y. Then., P. Y. Xue, J. P. Huang, J.
of
males
longer than Aedes
499 pp.
by females did not play as large

in survivorship as expected. Blood feeding or lack
Chinese with English summary).

Gillett, J. D.
1971.
Mosquitoes. Weidenfeld and
Nicholson Publishing Co., Inc., London, 274 pp.
thereofdid not affect survivorship ofEr. quinquevittatus

orAe. albopictus, but did
of Ae. aegypti and
significantly affect survivorship
Ae. bahamensis.
This report is the first life span or adult survivorship
Gilotra, S. K. and K. V. Shah. 1967. Laboratory studies

on transmission of Chikungunya by mosquitoes.
Am. J. Epidem. 86: 379- 385.
study of Ae. bahamensis. Survivorship for males and

females
of
this species was greater than that of Ae.
albopictus and
survived as
Ae.
long
aegypti.
as
Females
females
of
Er.
ofAe.
bahamensis
quinquevittatus.
If
Ae. bahamensis is an efficient vector, its longevity could
Gubler, D. J.
1970.
Comparison of reproductive
potentials of Aedes( Stegmoyia) albopictus Skuse
andAedes( stegomyia) polynesiensis Marks. Mosq.

News. 30: 201- 209.
make it a serious problem.

Our strain ofAe. aegypti behaved counter to reports
Hartberg, W. K. and W. H. Faircloth. 1983. Mitotic
in the literature( Christophers 1960, Gillett 1971). Our
chromosomes of the mosquito Eretmapodites quin-
males and
females
survived
longer
periods when under
quevittatus Theobald. Mosq. Syst. 15( 4): 325- 329.
the blood fed treatment, than when under the non- blood
fed treatment.
We have
no
explanation
for this
Hartberg, W. K. and E. J. Gerberg. 1971. Laboratory

Colonization of Aedes simpsoni ( Theobald) and
occurrence.
Eretmapodites quinquevittatus ( Theobald). Bull.
Wld. Hlth. Org. 45: 850- 852.

REFERENCES CITED
Ardoin, P. L. M. and D. I. H. Simpson. 1965. Relations
Hawley, W. A. 1988. The biology ofAedes albopictus.

J. Am. Mosq. Cont. Assoc. 4( suppl. 1): 40.
antigenique entre le virus Nyando- et deux virus

isoles
en
Ethiopie
partir
d' Eretmapodites. Bull. Soc. Path.
de
collectes
exot.
58: 574-
589.
Hylton, A. R.
1969.
Studies on longevity of adult
Eretmapoditeschrysogaster, Aedestogoi, andAedes

stegomyia) albopictus females. J. Med. Ent. 6:
147- 149.
Bauer, J. H. 1928. The transmission of yellow fever by

mosquitoes other than Aedes aegypti. Am. J. Trop.
Med 8: 261- 282.
and productivity of adult male and female Aedes
Brottes, H., A. Rickenbach, P. Bres, M. C. Williams, J.
J. Salaun,
and
Liles, J. N. and D. M. Delong. 1960. The longevity
L. Ferrara. 1969. A
new arbovirus
aegypti when reared separately and together on

three different diets. Ann. Ent. Soc. Am. 53: 277280.
prototype Okola virus YM-50- 64 isolated in the

Cameroons from
Eretmapodites
mosquitoes
chrysogaster.
human
new record-
Ann. Inst. Pasteur.
119: 543- 551.
Liu, Z. W., Y. J.
Zhang,
Population dynamics
and
of
Y. Z.
Aedes
Yang.
1985.
( Stegomyia)
albopictus ( Skuse) under laboratory conditions.

Acta Ent. Sinica 28: 274- 280 ( in Chinese with
Christophers, S. R.
1960.
Aedes
aegypti:
Its Life
English summary).
History, Bionomics, and Structure. Cambridge

Univ. Press Publishers, London.
Macnamara, F. N. 1953. The susceptibility
of chicks
to
DECEMBER, 1993
Semiliki forest
virus ( Kumba
Strain). Ann.
Trop.
Med. Parasit. 47: 9- 12.
113
Smithburn, K. C., A. J. Haddow, and J. D. Gillett. 1948.
Rift Valley fever. Isolation of the virus from wild

mosquitoes. Brit. J. of Exper. Path. 29: 107- 121.
McIntosh, B. M.,R. H. Kokemot, H. E. Paterson, and B.

De Meillon. 1961. Isolation
from four
of spondweni virus
species of culicine mosquitoes and a
report of two
laboratory infections with the virus. S.
Afr. Med. J. 35: 647- 650.
in
and
P. Neri. 1968. Studies on
Ethiopia
5.
Isolation
arthropod vectors.
877.
Aedes ( Howardina)
albonotatus (
Coquillet),
common domestic mosquito from the Bahamas.
Mosq. News. 25: 339- 343.
Serie, C., L. Andral, J. Casals, M. C. Williams, P.

Bres,
Spielman, A. and A. E. Weyer. 1965. Description of
of viral
Bull. Wld. Hith.
Worth, C. B., H. E. Patterson, and B. De Meillon. 1961.
yellow
fever
The incidence of arthropod-borne viruses in a
strains
from
population of culicine mosquitoes in Tongaland,
38: 873-
Union of South Africa. Am. J. Trop. Med. Hyg. 10:
Org.
583- 592.
DECEMBER, 1993
FLEAS FOUND ON MAMMALS IN SAN DIEGO COUNTY, CALIFORNIA
J. D. Lange
AB STRACT: As part of vector-borne disease surveillance activities, fleas were collected from sylvatic and domestic
mammals at 46 different localities throughout San Diego County from April, 1991, through December, 1992.
Twenty- four flea species( 12, 121 specimens) were collected from 26 of the 32 host taxa( 2, 397 specimens) examined.
Ten of these species have not been reported previously in the literature as occurring in San Diego County. A review
of unpublished flea surveys of this county yielded two additional species, Orchopeas leucopus and Pulex irritans,
which were not found during the present survey. Thirteen of the 26 flea species occurring in San Diego County have
been implicated as disease vectors. Oropsylla montana, the primary plague vector in the western United States,
comprised 62 percent of the specimens collected during this survey.
INTRODUCTION
activities, small mammals were sampled from April,

1991, through December, 1992, at 46 different localities
The importance
other
a number of
in California to determine the
relationships
between these
vectors and
domestic hosts. Surveys
situated at elevations ranging from 0 to 1, 575 m

throughout San Diego County. Eight of these localities
as vectors of plague and
diseases has been the impetus for
surveys
and
fleas
of
were each sampled monthly to determine seasonal

abundance offleas on specific rodents, while the majority
ecological
their sylvatic
of sylvatic mammals
have
of the other localities were sampled at least twice.
been conducted in the northern( Clover et al. 1989, Stark

and
Stewart
Schwan
et al.
and
and
Brennan 1957),
Evans 1941, Holdenried
and
Rutledge
as
1969, Jameson
Kinney
1979),
and southern (
Nelson 1983)
et
al.
1951,
Augustson 1943,
portions of the state, as well
in the Sierra Nevada Mountains ( Augustson 1942,
Murray
1971).
Fleas
also
have been
sylvatic and domestic mammals
collected
and
in
southern
12. 7 x 41 cm)( Tomahawk Live Trap Co., Tomahawk,

WI 54487).
Traps were baited with rolled oats and
placed in protected areas near active rodent burrows at
dusk and collected the following morning prior to 1000

hr.
The captured mammal was ushered into a clear
1957)
( Methoxyflurane)( Pitman- Moore, Inc., Mundelein, IL
et al.
1986). Flea
have been done in
et al.
surveys of
urban areas of
1938), Los Angeles( Schwan
60060).
A blood sample was drawn from selected
species for pathogen testing. The anesthetized animal
et
was combed for ectoparasites over a white porcelain pan
San Diego ( Augustson 1943, Prince
for approximately two minutes. Fleas were placed in

either two percent saline or 70 percent isopropyl alcohol,
The majority of these flea surveys have been either

duration, limited in number of hosts sampled, or
depending on whether pathogen testing was to be

conducted. The mammal was then placed in a trap, and
once revived, released in the vicinity of its capture site.
1985),
Eskey
and
1943).
of short
restricted to a
comparatively small geographical area.

This is particularly true for surveys of sylvatic mammals
carried out
present
of
7. 6 x 7. 6 x 25 cm) and 15 Tomahawk live traps( 12.7 x
California( Meyer and Holdenried 1949,
mammals
San Francisco(
Each locality was sampled using ten Sherman traps

(
plastic bag ( 3 ml x 38 x 61 cm) and anesthetized by

saturating the end of a cotton swab with Metofane
Ryckman 1971, Nelson
domestic
from
coexisting in interfacial
habitats in the San Francisco Bay area( Miles
al.
central
fleas
County
in
survey
southern
contributes to the
on sylvatic and
and
California.
domestic
identifies fleas
control purposes
during
Consequently,
Fleas and other ectoparasites were identified in the
the
laboratory as to species or subspecies, sex, stage( where
biological knowledge
appropriate), and number. Mammals were identified to
rodents
and their
in San Diego
flea-borne disease
species or subspecies based on the listing compiled by
possible
Bond( 1977). Eight domestic dogs from Barona and Las
outbreak.
Viejas Indian Reservations, a bat, and a deer shot at
hosts for
Camp
Pendleton
were also examined
for fleas.
All
ectoparasite and host data were recorded on a California

Mammal Collection and Ectoparasite Identification
As
part of plague and
Lyme disease
surveillance
Record Form.
1Vector Surveillance and Control Division, San Diego County Department of Health Services, San Diego, CA
92186, U. S. A.
115
DECEMBER, 1993
is characterized by coniferous forest.
RESULTS AND DISCUSSION
Orchopeas latens ranges from north- central

A
12, 122 flea
total of
identified from 26
species, was
during
following
California to San Diego County and parasitizes the gray
representing 24
Sciurus
Eads
1985).
A casual
33 host taxa
squirrel,
found
on the
recovery on S. b.( nudipes) was reported in the City of
of the mammals examined (
2, 396).
include
results
the more pertinent flea
of the
Fleas
the sampling period.
majority ( 99. 6%)
The
specimens,
were
San Diego ( Augustson 1943).
found in San Diego
species
et al.
During the present
survey, a male of 0. latens occurred on S. g. anthonyi

sampled at Green Valley Falls Campground( 1, 183 m),
for
unpublished records
griseus(
Cuyamaca Rancho State Park.
County.
This campground is
predominated by mixed woodland.
Malareus telchinus commonly occurs in most

western states including central and southern California
Ceratophyllid Fleas
The ground squirrel flea, Oropsylla montana, ranges

from Colorado
New Mexico
and
parts of Oregon and
west to
California
and
Washington( Hubbard 1947, Lewis
1988). In California, this cold- weather flea prefers
et al.
Spermophilus beecheyi( Linsdale 1946, Hubbard 1947,

Holdenried
Nelson
for
elevations(
Lang
Evans
United States ( Clover
western
Brennan 1957, Lewis et al. 1988). In San Diego County,

M. telchinus occurred on Peromyscus californicus
Del Mar ( Augustson 1943), and on
( insignis),
largest
majority being from S. b. nudipes(

also the dominant flea found at the higher
North America, the
dine
with
0.
s. sexdentatus
Riverside Counties ( Augustson 1943),
County
Park( 1, 575
record
the
and
while
and
San
were suggested to
specimens
0.
of
present
survey
This flea was
October
The majority
of
more common during the winter months.

Malareus sinomus also occurs in arid portions of
western North America, but prefers elevations between
the
700 and 2,000 m, with most records being from the

higher
ranges (
Haddow
et al.
1983).
It has not been
reported in Washington and only twice in Oregon
during
the
months
of
January.
western
states,
the deer
sporadic
mouse
flea,
distribution
with
was found naturally infected with

in New Mexico( Traub et al. 1983). Although 0.
Augustson 1943). It
leucopus was not encountered during the present survey,
be more common in the Southwest with southern

California records from Santa Barbara, Los Angeles,
and
Riverside Counties ( Augustson
1943).
An
unpublished San Diego County record gives a single

specimen from P. eremicus ( fraterculus) sampled in
Mataguay Creek ( 971 m), which is

characterized by grassland and chamise chaparral.
June, 1982,
at
Plague bacilli were found in specimens of M. sinomus
collected from three Peromyscus spp.( 11 specimens) in
three
Utah( Marchette et al. 1962). AlthoughM. sinomus may
from two Peromyscus eremicus( fraterculus)
not be an efficient vector among deer mice, it also may

be able to transmit the infection by mass infestation
an unpublished
examined
This flea was somewhat
in
Californiarecords being from Riverside County
specimens
fromNeotoma spp.( TABLE 1).
( Hubbard 1947, Lewis et al. 1988). This flea appears to
during
Hubbard 1947, Lewis et al. 1988, Hubbard 1949),
plague
survey, 70 percent of the specimens of M. telchinus

collected were from Peromyscus spp. and 22 percent
the
sexdentatus collected
Orchopeas leucopus, has
southern
may be an inefficient plague vector( Burroughs 1944,
from Neotoma lepida intermedia.
more common
through
In the
s.
were
Nelson 1983). Although M. telchinus
be
amplifying complex for epizootic plague

United States to include California and
Oregon ( Barnes 1982).
and
of fleas infesting a host thus increasing the chances of

infection ( Marchette et al. 1962). During the present
western
southern
Schwan
occurring in
1992). These
Nelson 1983). Orchopeas
its Neotoma hosts
m)(
Kartman and Prince 1956), it may effectively transmit

the infection via mass infestation, that is by large numbers
is from Palomar Mountain State
Schwan
m)(
sexdentatus and
Peromyscus spp., Palomar Mountain State Park( 1, 575
to have a
sexdentatus, appear
from Santa Barbara, Los Angeles,
are
a principal
elevations.
subspecies of the
southern California( Lewis, pers. commun.,
Diego
number of
the present
flea, Orchopeas
more common during the winter months( Jameson and
in California( Nelson 1980).
survey, with the

TABLE 1). It was
during
western
et al.
sylvatic
montana comprised the
north- south
et al.
Wills 1991).
and
its host, S. beecheyi, is the primary
collected
records
Surveys in
on arvicolids than Peromyscus spp. This flea was also
in the
Oropsylla
wood rat
1947).
is considered the most important sylvatic vector
while
In
and their carnivores ( Hubbard
Oregon ( Lewis et al. 1988) and San Francisco Bay
1986,
rodent involved with plague
fleas
spp.
( Miles et al. 1957) bothfoundM. telchinusmorecommon
plague
1989),
is the dominant flea found
but is also frequently found on

arvicolids, such as Microtus spp. and Clethrionomys
Peromyscus spp.,
1943,
higher
at the
et al.
montana
and
Hubbard 1947), with a preference for mesic niches in
these arid areas( Lewis et al. 1988). It usually occurs on
Oropsylla
host
on this
1951),
et al.
San Diego
in March, 1961,
County record reports

near
Julian( 1, 274
m) which
116
6)
sninosnw
DECEMBER, 1993
snyy
N
O
i6) sttllva snllvg
ci,
1) vuviui8.nn siydiapij
V
z
1) i Cuoyluv
0,
1)
suvou8lu
snau8
aviloq
s(
sninios
uiouioy,L
8
snoivao
vin vo snlo
oi
mm
thwoluoponyiiag
sisuaiiuvw ian.q snas,(wo.[ad
L) t3ai p 1 iouvs
to
r,
a)snpnvoquoi
spoiv8aw
o`
r,
e
a
L)
1 (
aiOJ
i ( oq snoswoad
oz(*
isuaydais snuuo snos( woad
as
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c,
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iiiagwv8 snivinoiurnu
1'
601) sninalaivrfsnoiwaia
snos(
snosLCwo1ad
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11
9)
su8su
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snosCwoiad
c)
In
en
en
Lb) silolovw sadWsnJ vwoloaN
a
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'
i7
t0) ddsgns eindai tnuoloaN
..
1 L.)
i
6S0`i) sad pnu i( ayoaaq sniiydauaads
K =
ra
t7
r
N
v
O
C
8V
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tZ
wi
_
i
1
y
2.
000
o
43
O
y
E-
C7
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Vg g
az
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Marchette
1962).
et al.
Peromyscus
During
M.
of
specimen
single
117
DECEMBER, 1993
the present survey, a
from
collected
was
sinomus
in June,
maniculatus gambelii examined
Pollitzer 1954). During the current survey, all specimens

of N.fasciatus were from R. rattus examined in urban
areas of San Diego County.
1991, at Lost Valley( 1, 031 m) which is characterized

by
mixed chaparral and coniferous
The deer
mouse
into
wagneri ophidius,
Cascade- Sierra Nevada Mountains
occurs west of the
and
Pulicid Fleas
forest.
flea, Aetheca
The
California ( Hubbard 1947).
southern
found to vary locally,

found on its normal host, P. maniculatus, in
A.
abundance of
wagneri was
rarely being
Lava Beds National Monument, Siskiyou
California( Stark and
northern
California
records are
Kinney
from Santa Barbara, Riverside,
Los Angeles( Augustson 1943),
10
report
fraterculus)
1988). The
be
scarce
present
flea
were
with
Most( 11)
the
also show that
involved with epizootic plague amplification in

California, Oregon, and western Nevada( B arees 1982).
w. ophidius to
Hoplopsyllus anomalus was the second most common
majority( 9
species encountered during the present survey( TABLE
of
which
m)
San Francisco
also
months of the year.
Echinophaga gallinacea is widely distributed

throughout the wanner regions of the world( Lewis et al.
ranges
from the
into Nevada
Bay
and
has been found
other
on
1988).
Although mainly a parasite of gallinaceous
birds, it also occurs on larger insectivores, large

carnivores, and rodents, such as ground squirrels( Lewis
1972).
Southern California records are from various
hosts in Kern, Santa Barbara, Los Angeles, and Riverside
The only southern California record is a

from Thomomys bottae, Los Angeles
Counties( Augustson 1943). San Diego County records

are from S. b. nudipes and a ring-neck pheasant,
During the present survey,
female of D. bluei were
Observatory
predominated
by
collected
from a T. b.
Campground ( 1, 477
Mountain ( TABLE
1).
This
Mus
Phasianus torquat, in Oceanside( Augustson 1943). E.
nigricans
gallinacea has been shown to transmit murine typhus
Palomar
under both laboratory and natural conditions( Hubbard
m),
is
Hubbard 1947).
occurs on
Rattus
on
other
occasionally
In Los Angeles County, N.
and
was the second most common
flea found
on
index( 2. 2) occurring
Schwan et al. 1985). In
the highest mean
in late spring and early summer(

the City of San Diego, it occurred
Augustson 1943),
on
Rattus
while an unpublished
1947).
In the present survey, most specimens occurred
on S. b. nudipes, followed by R. rattus( TABLE 2).
mixed woodland and grassland.
musculus,
with
a male and
campground
Nosopsyllus fasciatus commonly
Rattusrattus
The majority of the specimens occurred on S. b.
Hubbard 1947, Rutledge
Hubbard 1947).
fasciatus
2).
in San Diego County.
single specimen
and
1943), Kern
w. ophidius varies
A.
mixed chaparral
Thomomys spp., but
spp.
et al.
more common at the lower elevations during the warmer
rodents and sylvatic carnivores(
rodents (
in Alameda ( Meyer
et al.
Lewis
Arizona( Hubbard 1947). Its preferred hosts are gophers,
from
plague
by grassland and mixed chaparral. This

during November and December, 1992.
flea, Dactylopsylla bluei,
1979).
with
nudipes and averaged 2. 5 per host. This flea was also
south and east of
et al.
Rutledge et al. 1979).
is
locally, preferring
rare
1971,
and its S. beecheyi host, form a primary complex
specimens of this
from Lake Morena ( 934
These fmdings
The
Ryckman
in main-
County
found
was
1951,
Hoplopsyllus anomalus has been foundnaturally infected
Mataguay
at
a minor role
found A.
also
County,
m. gambelii.
characterized
flea
survey
warmer, drier months of the year ( Holdenried et al.
eremicus
P.
small rodent populations(
in San Diego
13) from P.
may play
common on S. beecheyi at lower elevations during the
( Evans et al. 1943), and Ventura Counties( Meyer and

Holdenried 1949). This flea, together with O. montana
six
in June, 1982,
wagneri
in
plague
from
specimens
examined
Creek. Aetheca
taming
Orange Counties
and
data). Unpublished San Diego
unpublished
records
County,
1969). Southern
In the United States, the ground squirrel flea,

Hoplopsyllus anomalus, ranges from southern Colorado
into eastern Oregon, western Nevada, and California
( Hubbard 1947, Lewis et al. 1988) where it is more
norvegicus
1987 survey
Cediopsylla inaequalis interrupta mainly occurs

west of the Cascade Mountains of Washington and
Oregon, with records also from Nevada, Idaho, and
California( Lewis et al. 1988). More frequent hosts are
rabbits and hares, particularly nest building Sylvilagus

spp. ( Lewis
et
al.
1988).
In southern California, it
occurred on Lepus californicus, Sylvilagus spp, a gray

fox, Urocyon cinereoargenteus californicus, and a
Although N.fasciatus
bobcat, Lynx rufus californicus, in Los Angeles County,

and on an opossum, Didelphis virginiana, at Point
Loma, San Diego County ( Augustson 1943).
is
may cause some human plague cases when the index
maintain
or
to
initiate
epizootics
high, it was unlikely to
Unpublished San Diego County records report it from

Sylvilagus auduboni ( sanctidiegi), City of San Diego
also
found it
each on
and on
common on
M. musculus( 1
S. beecheyi(
the disease in rat
R.
rattus and occurred once
examined),
nudipes)(
1).
populations
Neotoma lepida( 2),
for any length
of
time
and
Rancho Bernardo, from
a coyote,
Canis latrans
118
DECEMBER, 1993
zzz
8)
sun
runf s uvd
U
0
cu
06) snIWI
oo
snllvd
I) vivur8inf snuorway snapoaopp
0\
I)
I)
vuvnui8.nn snyd/
ap! Q
suaasvrau1a
iumuyavq sn8vpigS
enn Cl
a0
zz
i8apuauvs ruognpnv sn8vju/(
o\
en
O's
ra
v,
pnauuaq
snauroflva
snd7
0
y
8'
K,
i)!( aIMor I,toq snasiculoJad
r"
0
E
b91)
cv
P
U
00
swu8, sus snazuiofrJva snas,(

ruoJad
4.
0) ddsgns vprdaa vuIoJoaN
Lb)
sporavw
sadrasnf vruoloaN
a
0
000
8
ar
1(
6 SO` I)
sadpnu!(
ayaaagsnyydouuads
zU
y
x:
en
y
ea
4.
0
1
U
V
nz' ^ 3^'
zs
CI
00
8v/
C `
U
6
a
a a 414
cI '
i
4"
C'
r r
gg
Z' WC3WrY CI C) .
G/
3 3 o VI
s
a ySo
48
O- '.
Z {_
ri
s =
am.+
C
1 ,.
41 '.:
R.
localities. San Diego Zoo records two specimens from
San Diego Zoo. Cediopsylainaequalisinterrupta
two R. rattus, and five from a snow leopard, Panthera
shown to transmit plague or tularemia,
La Jolla,
clepticus),
rattus,
has
been
not
lagomorphs ( Lewis
survey,
and one specimen each on two
the present
uncia. Ctenocephalides f.felis has been shown to be

naturally infected with Rickettsia typhi, and may play a
significant role in transmitting this disease to humans
flea were collected from
( Irons et al. 1944). It was also suggested that some of the
foxi,
human cases of murine typhus occurring in Los Angeles

and Orange Counties were caused by cat fleas and their
both diseases have been
although
S.
1988).
et al.
most specimens of this
from
reported
During
a. sanctidiegi.
Another
flea, Euhoplopsylla
rabbit
occurs
southern
California being
cinerascens,
arizonae,
Riverside
San
Los Angeles
of this
specimens
flea
from Sylvilagus spp.,
were
during
collected
An
et al. 1970).
The capacity for C. f.felis to
transmit plague is probably negligible ( Lewis et al.
1988). Most( 11 of 17) specimens of C.f.fells collected
S.
during the present survey came from six of the eight
S.
and
on
( Adams
a.
dogs examined( TABLE 2).
Most
Rancho Bernardo.
auduboni ( sanctidiegi),
opossum hosts which were seropositive for R. typhi
with
from Sylvilagus
records
County,
County ( Augustson 1943).
Diego County record reports it
bachmani
unpublished
glacialis
in California ( Hubbard 1947),
primarily
survey
The dog flea, Ctenopcephalides canis, like C. f.
the current
felis, has a fairly general distribution throughout the
two from L. c.
with
western states, but is not as abundant as the latter
bennettii.
In North America, Pulex
be
associations and
P.
example,
and S. b. nudipes( Augustson 1943). The present survey
of other
found two specimens of C. canis on two of the dogs
most abundant species
examined. These dogs probably became infested from
a wide
also occurs on
variety
sylvatic canids because the dogs were not confined.
occurring on canids in the southwestern states( Hopla

1980). Pulex irritans, on the other hand, occurs mostly
humans( Hopla 1980),
on canids,
and swine(
Both fleas have been found
1985).
Leptopsyllid Fleas
Holland
on mule
Leptopsylla segnis commonly occurs on Rattus
deer in
California( Hopla1980). Unpublished San Diego County

records report P. simulans on
City
of
San Diego,
spp.
La Jolla,
from San Diego
and the other
County
Pulex
identified
were
and on
among Cynomys populations

Hopla 1980). Although P. irritans is presumably of no
important
plague vector
Lewis
et al.
1988).
In Los
City of San Diego, and on R. norvegicus, Chula Vista

(
Prince 1943).
An unpublished San Diego County
survey also found L. segnis to be the most common flea

on R. rattus in urban areas. Other unpublished records
for the City of San Diego report it as occurring once on
were
R. norvegicus, S. b. nudipes, N. 1. intermedia, and on an
from dogs in New Mexico ( Hopla 1980).
opossum. Although L. segnis rarely feeds on humans
as a vector
plague- positive
collected
musculus (
Smit
importance
1958),
M.
Diego County, it was found on R. rattus in Jamul and the
specimens
as simulans or
irritans by the late B. Nelson. Pulex simulans may be an
and
Angeles County, L. segnis was the most widespread and

abundant flea collected from R. rattus, being more
common from December through February with the
highest index being 4.4( Schwan et al. 1985). In San
16 of 28 opossums, mostly
on a coyote,
three gray foxes from different localities. Two female

P. irritans were found on two R. rattus examined at the
San Diego Zoo. These
Hubbard 1947). Ctenopcephalides canis occurs mostly
on sylvatic canids, especially Canis and Vulpes( Lewis

1988). In San Diego County, it was found on a human
both host
as to
variety of habitats( Hopla 1980). For

is primarily found on colonial
Smit 1958). It is the
mammals(
simulans
but
rodents,
simulans was reported to
Pulex irritans
more adaptable than
in the
119
DECEMBER, 1993
among
sylvatic rodents (
specimens
of this
flea
Pulex irritans also has been implicated in human

cases reported
1980). The
from
present
various parts of
plague
the world(
Hopla
survey found most specimens of P.

Pulex
dogs examined.
and is therefore probably not responsible for human

cases of murine typhus; it may vector this infection
between rats( Traub et al. 1978). This flea is not a vector
Pollitzer 1954).
The current survey found
simulans on two of the eight
of plague (
irritans
most specimens of L. segnis on R. rattus.

Peromyscopsylla hesperomys pacifica was found
was not collected.
The
found
cat
flea, Ctenocephalides f.fells, is generally

United States ( Hubbard 1947).
throughout the
to occurcommonly in the PacificNorthwestandnorthem
host is Felis domestica, it readily

domestic dogs, Can us familiaris, and is
California, and the eastern portion ofthese states lacking
occasionally found on other animals( Augustson 1943,

Hubbard 1947). Unpublished San Diego County records
survey in the same area found it uncommon( Lewis et al.
City of San Diego,
maniculatus( Hubbard 1947). An Oregon survey found
Although its
attacks
report
normal
it on 40
of 41 opossums
and on a coyote and three
in the
gray foxes from
various
distinct desert
conditions (
Hubbard 1947).
Another
1988). Preferred hosts are Peromyscus spp., particularly

it in August
and
September,
with a single specimen
120
being
in December( Lewis et al. 1988). Both

Californiarecords are from PalomarMountain.
Counties( Augustson 1943). During the present survey,
found on four Peromyscus spp.,
Perognathus spp.( TABLE 3). This flea occurred from
collected
southern
Fifteen
specimens were
Palomar Mountain State Park ( Schwan
1983),
while an
1981
unpublished
and
The
m).
found P. h.
specimens(
P.
of 44)
Nelson
October through April, with 90 percent ofthe specimens
being recovered from October through February.
Crestline
to be uncommon, with most
The nest flea, Catallagia luski, was described from

Peromyscus boylii ( rowleyi) collected at Palomar
Mountain State Park ( Schwan and Nelson 1983).
from Peromyscus spp., especially
Previously unidentified specimens from other California
San Diego
present
pacifica
41
most specimens of C. carteri were also found on two
record reports a
single specimen from P. eremicus( fraterculus),
900
DECEMBER, 1993
County
survey
also
insignis( TABLE 3). This flea was also

more abundant from August through the winter months
californicus
localities were also identified as C. luski( Schwan and

Based on these collections, this flea
Nelson 1983).
occurring in January.
Odontopsylla dentatus occurs from the Rocky
Mountains to the Pacific Coast States( Hubbard 1947).
Its preferred hosts are rabbits and hares and is also found
appears to occur above 1, 000 m in the western Great

Basin and in the southern Sierra Nevada, Transverse,
and Peninsular Mountain ranges of California( Schwan
on their predators,
weather flea since it was four times more abundant on P.
with peak numbers
and
Nelson 1983).
Catallagia luski may be a cold-
especially bobcat and coyote( Lewis

et al. 1988). Southern California records report it from
Sylvilagus spp. and agray fox, Los Angeles County, and
boylii ( rowleyi) during November than the previous
from L. c. richardsoni andN.fuscipes, Riverside
Catallagia species prefer cricetine and microtine rodents
Augustson 1943). An
record reports
with
a single specimen
Diego Zoo.
of
unpublished
it from S. a.
During
0. dentatus
were
San
sanctidiegi, Rancho
occurring
the present
found
on a
on
March ( Schwan
County
Diego County
R.
Utah,
southeastern
1977).
occurs
well as
Diego
California
1988), they are more often found on Spermophilus spp.
San
than other mouse fleas, and thus may play an important

role in transmitting plague to amplifying hosts such as
rattus,
survey, two specimens

S. b. cinerascens, Lake
to southern
east
and southern
Southern California
S. lateralis( Wilson, pers. commun., 1991). The current

survey found one specimen of C. luski on S. b. nudipes
collected in April, 1991, at Green Valley Falls

Campground( TABLE 3).
Atyphloceras m. multidentatus occurs in the Pacific
Nevada,
Arizona( Barnes
records
Coast States( Hubbard 1947, Lewis et al. 1988), being
et al.
a common winter flea found mostly on Microtus,
indicate that it
Peromyscus( Lewis 1974), and Neotoma( Hubbard 1947)

in the Pacific Northwest. In northern California, it was
mostly in coastal chaparral and oak woodland as

in typical desert habitat( Barnes et al. 1977). San
County
records are
from
nests of
N. fuscipes
also common during the winter in Plumas County

( Jameson
Neotoma spp. examined near Banner and

Julian, Ramona, Guatay, and the
City of San Diego
macrotis) or
Barnes
et al.
collected
1977). Most
A.
generally
agree
Lewis
et al.
that A.
1988),
nudatus
current
Kinney 1969). In southern California, A. m.

multidentatus was collected from P. eremicus, Los
and
Angeles County, and from a harvest mouse, Reithro-
dontomysmegalotis, San Bernardino County( Augustson

1943). This flea occurred on Peromyscus spp. at Palomar
findings
subspecies
Mountain State Park ( Schwan and Nelson 1983), and

was found naturally infected with plague, although
are
infrequently
al.
found on non- Neotoma hosts ( Barnes et

1977). The present survey found this flea to be more
common on
hosts
during
natural transmission of this disease remains to be

demonstrated( Lewis
the winter and spring months.
are
1988). Southern California records

from Santa Barbara, Los Angeles, and Riverside
et al.
The present survey
from October through January.

The kangaroo rat flea, Meringis cummingi, is fairly
record
1943, Lewis
1988).
the 17 specimens were collected from Neotoma spp.
Monterey County to southern California with one
from Nevada( Lewis, pers. commun. 1993). This

apparently cold- weather flea prefers pocket mice,
Perognathus spp., particularly californicus( Augustson
et al.
also found A. m. multidentatus uncommon. Most of
The rare flea, Carteretta carteri, has been recorded

from
Brennan 1957), although only six
from February through June in Siskiyou County( Stark
specimens of A. n. nudatus
during the present survey were from Neotoma
n. nudatus(
and
specimens were found on 205 P. maniculatus examined
spp.( TABLE 3). Although Peromyscustrueimartirensis

tends to occupy Neotoma lodges( Jameson and Peeters
1988) and should have thus yielded more specimens
of
Although most
( Hubbard 1947, Eads and Campos 1979, Lewis et al.
Hystrichopsyllid Fleas
The wood rat flea, Anomiopsyllus n. nudatus, ranges
southern
Nelson 1983).
Bernardo,
Morena.
from
and
common in southern Oregon, California, and Nevada

(
Lewis, pers. commun. 1993). In southern California, it

was the most common flea found on kangaroo rats in the
coastal areas of Santa Barbara and Los Angeles Counties
121
DECEMBER, 1993
i)
snalwofrlva a spo
Cyy
8
N
6)
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snyy
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i 6) snllve snnvd
tO
suaasveaula
1)
Z)
luvunpvq sn8nilnl'CS
suvinwls slll8v s(
wopodrq
cin
Si) sllviouuaf snalwofilva sngrou8oead
xvllvf fsnglvu8oead
C
Elk)
co
E
a) snpnvat8uol
spop8aui
skwoiuopoegnaJ
r;
L) slsuaautvw lapel snas,(woead
1)
lCa7Moa
miCoq snadwoead
Cl
i
yy)
lsuagdais snpuua snas(
woead
w;
Co
08)
nlaqum8 snvlauvw
snaswwad
CS
601) snlnaearoef snalwaea
snas
Cwoead
o
vi
o
0c
91)
slulsur
b0)' ddsgns vpldal
o.
Lb)
swaavw
vwo
z
oj
snalwof lva snaswoead
oal l
sadiasnf vwoJoaNN
en
4.
4.
ci.
3i
dU
6"
0
tsR
r(
6S0` i)
sadrpnu!
Kagaaaq
snlrgdouuadg
0.
N. N
to
y
i
F3
1 .. 1,
4.
c-
a .E .
O O p
g.
L.1sod
4.
332 ",
122
Augustson 1943).
Unpublished San Diego
County
DECEMBER, 1993
311- 318.
records give one specimen from Dipodomys agilis

cabezonae)
During
examined
in March, 1961,
near
Julian.
the present
cummingi were
survey, only two specimens of M.

collected, both from Lake Morena, with
Augustson, G. F. 1942. Ectoparasite- host records from

the Sierran region of east-central California. Bull.
S. Calif. Acad. Sci. 40: 147- 157.
one being from D. a. simulans and the other from P. m.

gambelii.
Augustson, G. F.
1943.
Preliminary records and
discussion of some species of Siphonaptera from

Ischnopsyllid Fleas
the Pacific Southwest. Bull. S. Calif. Acad. Sci. 42:
The bat flea, Myodopsylla
gentilis, ranges
from
the Rocky Mountains to the Pacific Ocean ( Hubbard

1947), with preferred hosts being Myotis spp.( Hubbard
1947, Lewis
flea
et al.
1988). It is
endemic to the
1988).
the most common
Pacific Northwest ( Lewis
It is apparently less
California,
with a single record
occulatus,
Riverside
common
in
bat
et al.
southern
being reported from M.

County ( Augustson 1943). The
69- 89.
Barnes, A. M., V. J. Tipton, and J. A. Wildie. 1977. The

subfamily Anomiopsyllinae ( Hystrichopsyllidae:
Siphonaptera). I.
A revision of the genus
Anomiopsyllus Baker. Great Basin Nat. 37: 138206.
survey found six specimens( 1 male, 5 females)

from a Myotis c. californicus, found in
August, 1992, near Fallbrook.
Barnes, A. M.
Mammals Not
Bond, S. I. 1977. An annotated list of the mammals of
current
of
M.
not
gentilis
Yielding
Fleas
The following taxa examined during the

survey did
yield fleas: Tamias m. merriami ( 1 specimen);
1982.
Surveillance and control of
bubonic plague in the United States. Symp. Zoo.

Soc. Lond. 50: 237- 270.
San Diego County, California. Trans. San Diego

Soc. Nat. Hist. 18: 229- 247.
Perognathuspenicillatusaugustirostris( 2); Chaetodipus

baileyi hueyi( 2); Perognathus longimembris pacificus
1); Chaetodipus spinatus rufescens ( 1); Thomomys
bottae sanctidiegi( 1); and Rattus norvegicus( 2).
Burroughs, A. L. 1944. The flea Malareus telchinum,

avectorofPasteurellapestis. Proc. Soc. Exp. Biol.
Med. 55: 10- 11.
Clover, J. R., T. D. Hofstra, B. G. Kuluris, M. T.

Acknowledgments
Schroeder, B. C. Nelson, A. M. Barnes, and R. G.

Botzler.
wish
to thank the
confirmed or identified
following
individuals
who
fleas collected during this survey:
Dr. Robert E. Lewis, Department
Entomology, Iowa
State University, Ames, Iowa; Dr. Edwardo G. Campos
of
Plague Branch, Centers for Disease Control,

Fort Collins, Colorado, and Barbara Wilson,
Environmental Management Branch, California
Department of Health Services. I also thank Janet Ortiz,
retired),
1989.
Serologic evidence of Yersenia

pestis infection in small mammals and bears from
a temperate rain forest of north coastal California.
J. Wildl. Dis. 25: 52- 60.
Eads, R. B. and E. G. Campos. 1979. Description of a

species of Catallagia ( Siphonaptera:
Hystrichopsyllidae: Neopsyllinae). J. Med.
Entomol. 16: 291- 294.
new
Program Manager, for her full support during the course

of
this survey, the
Surveillance Program staff for their

initiative, Rene Rada who helped with the
and Gerri Leviston who typed the tables.
efforts and
revisions,
Lastly,
I dedicate this
work to
the late Jim
Shoemake,
Eads, R. B., B. C. Nelson, G. O. Maupin, and A. M.

Barnes. 1985. Orchopeas fleas ( Siphonaptera:
Ceratophyllidae) of the western gray squirrel,

Sciurus griseus. J. Med. Entomol. 22: 630- 636.
Program Supervisor, who helped to make this survey

possible.
Eskey, C. R. 1938. Flea infestation of domestic rats in

San Francisco, California. Publ. Hlth. Rpt. 53: 948REFERENCES CITED
Adams, W. H., R. W. Emmons, and J. E. Brooks. 1970.

The changing ecology of murine( endemic) typhus
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951.
Evans, F. C., C. M. Wheeler, and J. R. Douglas. 1943.

Sylvatic plague studies. III. An epizootic of plague
among
ground
squirrels (
Citellus beecheyl) in
Kern
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Holland, G. P. 1985. The fleas
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of the Pacific Northwest. Oregon State Univ. Press,
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Hubbard, C. A. 1947. Fleas of western North America.
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wild animals in Utah. IV. A wild rodent
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Hubbard, C. A.
Marchette, N. J., J. B. Bushman, D. P. Parker, and E. E.
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Jawetz.
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plague
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IV.
Inapparent latent sylvatic plague in ground squirrels

Irons, J. V., S. W. Bohls, D. C. Thurman, Jr.,
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Probable role of the cat
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murine typhus.
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Meyer, K. F. and R. Holdenried. 1949. Rodents and

fleas in a plague epizootic in a rural area of
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H. J. Peeters. 1988. California
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California, with special reference to plague. Am. J.
Trop. Med. Hyg. 6: 752-760.
Calif. Press, Berkeley, Calif., and Univ. Calif.
Murray, K. F. 1971. Epizootic plague in California,
Press, Ltd., London, England, 403 pp.
1965- 1968. Unpub. Rpt. Calif. Dept. Health, Bur.

Kartman, L.
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F. M. Prince.
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Vector Contr., 76 pp.
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Nelson, B. C. 1980. Plague

roles
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Schwan, T. G., D. Thompson, and B. C. Nelson. 1985.
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Fleas on roof rats in six areas of Los Angeles
Proc. Vert. Pest.
County, California: their potential role in the
of sylvatic
ecology in California.
Conf. 9: 89- 96.
plague
rodents
transmission of plague and murine typhus to
Nelson, B. C., M. B. Maddon., and A.Tilzer. 1986. The

interface among domestic/ wild
fleas, pets, and man in urban plague
ecology
in Los Angeles County, California. Proc. Vert. Pest
Conf. 12: 88- 96.
complexities at the
rodents,
Pollitzer, R. 1954. Plague. W.H. O. Monogr. Ser. No.

22, Geneva, Switzerland, 698
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of fleas on rats collected
in
humans. Am. J. Trop. Med. Hyg. 34: 372- 379.

Smit, F. G. A. M.
1958. A preliminary note on the
occurrence of Pulex irritans Linnaeus and Pulex

simulans Baker in North America. J. Parasit. 44:
523- 526.
Stark, H. E. and A. R. Kinney. 1969. Abundance of

rodents and fleas as related to plague in Lava Beds
National Monument, California. J. Med. Entomol.
6: 287- 294.
states west of the 102nd meridian and their relation

to the
dissemination
of plague.
Publ. Hlth. Rpt. 58:
700.
study of rodent and burrow flea populations. Proc.
Rutledge, L. C., M. A. Moussa, B. L. Zeller, and M. A.

Lawson.
1979.
Field
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Fort Hunter Liggett,

California. J. Med. Entomol. 15: 452- 458.
vectors of sylvatic plague at
Ryckman, R. E 1971. Plague vector

The role of climatic factors in
studies.
Part II.
determining seasonal
fluctuations of flea species associated with the

California ground squirrel. J. Med. Entomol. 8:
541- 549.
Catallagia luski ( Siphonaptera: Hystrichopsyllidae), a new
Soc. Exper. Biol. Med. 47: 140- 142.
Traub, R., C. L. Wisseman, Jr., and A. Farhang- Azad.

1978.
The ecology
of murine
typhus - a critical
review. Trop. Dis. Bull. 75: 237- 317.
Traub, R., M. Rothschild, and J. F. Haddow. 1983.
The Rothschild collection of fleas. The family

Ceratophyllidae: key to the genera and host
relationships, with notes on their evolution,
zoogeography and medical importance. Univ. Press,
Schwan, T. G. and B. C. Nelson. 1983. Description of
revised
Stewart, M. A. and F. C. Evans. 1941. A comparative
flea from California
mice, and a
key tamale Catallagia from North America.
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Cambridge, U. K. Distrib. by Academic Press, Inc.

London), 228 pp.
Wheeler, C. M. and J. R. Douglas. 1941. Transmission

studies of sylvatic plague. Proc. Soc. Exper. Biol.
and
Med. 47: 65- 66.
DECEMBER, 1993
ELEVATED DOSAGES OF BACILLUS THURINGIENSIS VAR.

ISRAELENSIS FAIL TO EXTEND CONTROL OF CULEX LARVAE
M. S. Mullal, J. D. Chaney1, and J. Rodcharoenl
ABSTRACT: Vectobac 12AS, a liquid formulation containing the microbial agent Bacillus thuringiensis var.
israelensis, has proven to be an effective larvicide for mosquito abatement. Like other B. t.i. formulations, the
duration of larvicidal activity is relatively short, usually lasting no more than a week. The studies reported here
examined the possibility of extending the period of efficacy for the control of Culex mosquitoes by increasing the
dosage to five and ten times the minimum effective rate. Dosages of 0.2, 1. 0, and 2. 0 kg/ ha of this B. t.i. formulation
were evaluated against natural mixed populations of Culex quinquefasciatus, Culex stigmatosoma, and Culex
tarsalis occurring in field ponds at two locations in southern California. The results showed that the excessive
Vectobac 12AS dosages did not significantly extend the duration of effectiveness for controlling mosquito larvae.
Pertinent to the outcome of these studies were two important natural regulating factors which also affected the
abundance of mosquito populations in the ponds: 1) a progressive reduction in ovipositional attractancy of the
breeding site and 2) an increasing incidence of predation by aquatic macro-invertebrates. As a result of these
interacting factors, the larval populations following treatment with the three dosages of Vectobac 12AS never
recovered, demonstrating that larvicides with a margin of safety to nontarget biota can achieve desirable results using
minimum effective dosages, negating the justification for using higher rates to extend control.
and repeated treatments. In view of the anticipated rise
INTRODUCTION
in cost due to the use of greater amounts of material per

Although
more
registered
ten
than
years
ago,
Bacillus thuringiensis var. israelensis( B. t. i.) continues
application, the resulting fewer required treatments

should offset this concern.
to be one of the most effective and dependable microbial

larvicides available for use in mosquito control programs.
Presently, due to the lack of the availability ofalternative

chemical and microbial agents,
being
made
to improve the
continuing
efforts are
Over the 1991- 1992 breeding seasons, four field
B. t. i. in
tests were conducted in experimental ponds at two
from,
Riverside campus, and the Aquatic and Vector Control
in
Research Facility in the Coachella Valley. The ponds at

these locations were nearly identical in size, 27 and 30
m2, respectively. Aside from the wanner conditions in
effectiveness of
southern California sites: the University of California,
mosquito control.
With
a good selection of products to choose
various commercial
B. t.i. formulations
operational control programs

provide
long- lasting
that assessed
control
various
control of mosquitoes.
In
studies
B. t. i. formulations, the duration
biweekly)
or
impoundment
and
of
and
commercial
it
was
longevity
B. t.i.
ten times the
low desert
region),
the major
Darwazeh 1985,
covered with various grasses ( primarily Bermuda)
extending over their sides and bottoms. The Riverside
than the
ponds contain dirt bottoms encompassed by wooden
desirable to investigate the
board siding and are kept devoid of vegetation ( Mulla
product.
is
much greater
higher dosages using
The
purpose of
this study
rates of application,
five
Entomology, University
of
et al. 1982).
To enhance mosquito oviposition, 2 kg of organic
and
material ( either chicken lay mash or rabbit pellets,
the
depending on availability) was added to each pond prior

to flooding.
Larval populations were monitored by
longevity of control, thus, reducing the need for frequent
of
Valley (
cost of treatment,
and
minimum effective dosage, will extend
Department
Coachella
vegetation. The ponds at the Coachella site are naturally
Mulla
of
determine if higher
the
difference between these two sites was presence of
control of asynchronous
Since the
1990).
and surveillance
cost of material,
efficacy
for the
mosquitoes(
Lacey
monitoring,
was to
but generally these do not
has been short, necessitating frequent applications
weekly
Lacey
are used now
California, Riverside, CA 92521, U.S. A.
126
using
400
capacity dipper. Five dip
ml
pond were taken, one

sample
located
observed
Only 3rd and 4th-instar counts were used in these
samples per
from each corner plus an additional
where the mosquito
to be concentrated.
immatures
Counts
DECEMBER, 1993
calculations because early instars, especially 1st, do not
were
provide a good measure of activity of the agent as they

may not have been exposed for sufficient time.
of sampled
immatures were divided into three categories: early( 1st

and
2nd- instars), late( 3rd
and
4th- instars),
and pupae.
Dip samples were taken before treatment, two to three

days
after,
and
minimum of
species
monitored
using
composition,
water
temperature
minimum- maximum
thermometer.
Treatments using the B. t. i. liquid formulation

Vectobac 12AS ( 1200 ITU/mg- Abbott Laboratories,
N. Chicago, IL), were initiated before the first cohort of
larvae began pupating. Three rates were used

four tests conducted: 0.2 ( minimum effective
mosquito
in the
rate),
1. 0,
and
2. 0
untreated controls.
used
in
until all
each test.
kg/ ha along with their respective

Depending on the number of ponds
four
available, three or
of
Percent reduction was

Mulla
et
al.
replicates per
1971)
treatment
the water temperature over a 28 day evaluation period

ranged from a mean minimum of 21. 8 C to a mean
maximum of
31. 3 C.
High densities of larvae were
present in all ponds prior to treatment, averaging 50 to

70 late instars
per
1).
dip ( Fig.
The control ponds
exhibited a stable larval population profile over the
course of the experiment, averaging 60 late instars per
dip initially and gradually dropping to 44 per dip a

month later.
Two days after treatment, an immediate drop in

mosquito densities was observed in all treatments. The
were
lowest dosage of0.2 kg/ haproduced 95 percent reduction

of larvae, whereas, 100 percent control was achieved at
B. t. i. activity had disappeared.
the 1. 0 and 2.0 kg/ha rates. Low numbers of late instars

began reappearing in the treatment ponds, regardless of
dosage, seven days after treatment. Overall, the reduced
larval densities persisted for the remainder of the test,
calculated
which
using Mulla' s formula
compensates
population changes reflected
for
in the untreated
natural
ponds.
The comparative efficacy of the treatments was evaluated
statistically by using
located at the Riverside facility in August 1991, where
populations were monitored
Larval
indications
The first trial was conducted in field test ponds
was collected on each
identified. Pond
and
basis
post- treatment
weekly
50 late instar larvae
sampling date
was
on
To determine
thereafter.
analysis of variance (
ANOVA).
with the exception of two mild resurgences in the

middle rate,
1 kg/ ha, treated
00.2
a 1.
ponds.
A statistical
kg/ ha
0 kg/ ha
80
2. 0 kg/ ha
Check
60
40
M
ven
O
20
bq
0
0
14
21
28
Days Post-Treatment
Figure 1.
Efficacy of various rates of Vectobac 12AS against Culex larvae at Riverside ponds( AugustSeptember 1991).
between the
comparison
the
of the B. t.i. application. Prior to treatment, the initial
significant difference
early ins tar densities averaged 82 larvae per dip then

dropped significantly in response to the Vectobac 12AS
post- treatment
highest and lowest rates showed no
densities
of
in efficacy. Both these treatments were significantly

different from the control population( ANOVA, P< 0.05)
Unlike the
results
for the highest
variability. There
and
lowest
only three per dip. Not until after day ten ( three days
after the addition of a secondary supplement of organic
material to increase oviposition) did the numbers of
rates,
0 kg/ ha) exhibited
the ponds treated at the middle rate( 1.

unexpected
application( all rates) and by day seven were averaging
post- treatment assessment period.
throughout the
was a slight resurgence
early instars begin to rise, an indication that the effect of

the treatment was wearing off. The decline in larvicidal
in
Fig. 1) reaching
than
those seen in
higher
that
were
levels
significantly
0. 001 and P
P
<
ANOVA,
rates
(
treatment
other
the
larval densities
days 10
on
had
the high
due to the
was reduced
in the
into these
B. t.i.
dip
per
dip)
middle rate pond,
per
counts
by
highest
was
following
highest
rates.
This
Throughout the assessment period of this
the high
control,
averaging
progressed,
these and
develop,
to
continued
days 10 and 21 (
resurgence seen at
experiment, the mosquito species composition of the
untreated ponds at Riverside was monitored using 4thinstar larvae to ensure accurate determinations. Initially
( samples taken eight days postflooding) Culex
quinquefasciatus
early
Fig.
and Culex stigmatosoma were the
predominant species, making up 56 and 40 percent,
respectively, with the remaining 4percent, Culextarsalis

(
Fig.
2).
As time progressed, populations of both Cx.
quinquefasciatus and Cx. stigmatosoma declined, while
those of Cx. tarsalis steadily increased. By day 15 postflooding, Cx. tarsalis had become the dominant species.
Subsequently, on day 22, Cx. quinquefasciatus
completely disappeared. At the end of the test period, 35
1).
Because the
lowest
of
warranted.
the
ponds remained
low for the
experiment, a second treatment was not
With few
there was
contributed
were
due to
extended
Vectobac 12AS treatments,
other
oviposition
factors
activity
consider.
invertebrates
other
little
decline in
evidence
that
present
predators
in the
ponds,
significantly
to the extended reduction. If high populations
of predators were present, the

check ponds would
not the case (
Fig.
1).
larval
populations
in the
have declined rapidly, which was

However, a drop in oviposition
in early
activity was indirectly confirmed by a reduction
instar larvae in the check ponds. Initial samples averaged
100 young larvae per dip but dropped quickly, declining

markedly two days post- treatment to an average of 38
dip
leveling
for the
omitted).
entirely
of
Cx. tarsalis( 99%)
with an occasional
Cx. stigmatosoma( 1%).
In the second trial, conducted at the Coachella
of the
and
almost
Valley facility in late October 1991, the water

temperatures during the course of the test ranged from
important to
larvae
days postflooding, the larval composition consisted
larvicidal
densities
such as predation and a

were
late instars in both the
Although it appears that the sustained reduced
population
activity
populations of
highest rate
and the
duration
per
and
kg/ha.
lowest
and
similarly for the normal

flow ponds in the
and
subsequent unaffected cohorts
producing the
This
days
two
the
As time
dip.
in this
concentration
treatment showed considerably less
38 early instars
in both the lowest
more effective than the minimum effective rate of 0. 2
by
ponds.
complete reduction of
was produced
averaging 4
occurred
middle
indicated
to ineffective levels.
While nearly
instar presence
flow
in the
apparent
treatment.
activity, based on the observed increase of early instars,
flowing
flushing and dilution action of
the
water,
additional
rates(
replicated ponds
amounts of water
treatment
already
see
excessive percolation rates
Consequently,
the
21 (
and
clearly demonstrated that higher rates of B. t.i. were no
0. 01, respectively).
Two of the three
treatment
127
DECEMBER, 1993
Such
off
rest of
a
27 early instars
the assessment period ( data
to an average of
significant
impossible for the late instar

their original
drop
would
make
it
populations to recover to
levels.
in the early instar counts of the

treatment ponds were important in assessing the efficacy
Changes
observed
a mean minimum of 15. 9 C to a mean maximum of

27. 6 C. Pretreatment late instar counts averaged 73
larvae per dip. Assessment of this test lasted only two

weeks due to the rapid decline of larvae. Three days
following treatment, all rates of application, 0.2, 1. 0,

and 2.0 kg/ ha, yielded excellent control of 97, 100, and
100 percent, respectively( Fig. 3). The larval populations
in the untreated control ponds, immediately began to
decline and by seven days post-treatment, the check
samples were averaging only six late instars per dip, not
significantly different from the three B. t.i. treatments.
As time progressed, the densities continued to drop and
on day 14 the experiment was terminated.
Only the three day post- treatment larval densities in

the treatment ponds for all three rates were significantly
lower than those in the control ponds ( ANOVA, P
0. 01).
Shortly thereafter ( day 7) the control pond
counts dropped to levels similar to those in the treated

ponds
making it impossible to determine if B. t.i.-induced
128
DECEMBER, 1993
100
Ir
0.
60
40
g.
20
0
8
10
15
22
29
Days Post-Flooding
Figure 2.
Culex tarsalis
Ej Cx. stigmatosoma
Cx. quinquefaciatus
35
Succession of mosquito species( 4th-ins tar) in control ponds dueing first experiment at Riverside
August- September 1991).
100
80
0-
0. 2 kg/ ha
0-
1. 0 kg/ ha
2. 0 kg/ ha
i
60
Check
Y
b
M
40
q+
a
hiiiip,.
g 20
a
0
0
14
Days Post- Treatment

Figure 3.
Efficacy
of various rates of Vectobac 12AS against Culex larvae at Coachella Valley ponds( October
1991).
129
DECEMBER, 1993
larvicidal activity
There were
was still a
factor.
reasons
several
temperatures were
13. 1
and 20.2 C, respectively.
for the dramatic
Because of the shortness of the season, Vectobac 12AS
larval densities
was applied as soon as 3rd and 4th- instars began
reduction and continued suppression of
in the check ponds. Observations regarding the presence
appearing in the ponds. Pretreatment samples for all
of egg rafts and earlylarvalinstarsindicatedthatmosquito
ponds were similar, averaging 12 larvae ( late instars)
first few
primarily
to the onset of inclement weather rather than to a drop in
per dip( Fig. 4).

Very good control was achieved by day three,
where the 0.2, 1. 0, and 2. 0 kg/ ha rates produced 94, 97,
By day
and 99 percent larval reductions, respectively. Similar
important regulating
the presence of large numbers of
to the previous tests, the resulting reduced population

densities remained unchanged for the duration of the
oviposition
days
had declined markedly
of this test.
This reduction
the ovipositional attractancy
became
seven, predation also
factor as
evidenced
hydrophilid
and
by
during
the
was attributed
the
of
ponds.
an
dytiscid beetle larvae( data
omitted).
Changes in the larval composition monitored in the

Coachella
Valley ponds during this experiment showed
a similar trend observed
Initially,
ponds).
treatment,
6%), but
prevailed
Cx.
two
in the first
species
in the
two
weeks
Cx.
tarsalis
larvae
94%)
and
in November 1991
three weeks after treatment.
Statistical analysis of the results for trial three
proved to be unreliable because of the high degree of
ponds.
In the third trial located

started
Riverside
only Cx. tarsalis
quinquefasciatus(
after
trial (
were present prior to
test, showing no signs of recovery. However, the larval

densities in the control ponds continued to climb during
the first week, peaking at seven days post- treatment
with an average of 41 per dip, then rapidly declining to
insignificant levels, signaling the end of the assessment,
at the
under
Riverside
considerably
facility
cooler
conditions) the mean minimum and maximum water
variability of the data collected from the check pond

populations compounded by the inadequate number of
repetitions( only two ofthe three control ponds produced
50
0 0.2
o- 1.
kg/ ha
0 kg/ ha
40
9 2.0
kg/ ha
CD
Check
A-
30
M
n
w
20
10 :`
14
21
Figure 4.
Efficacy of various rates of Vectobac 12AS against Culex larvae at Riverside ponds( November
1991).
130
Even
sufficient mosquito populations).
so,
it is apparent
minimum of 19. 3 C to a mean maximum of 28.0 C.

The results of this trial are illustrated in Figure 5, where
Fig. 4) that all of the application rates evaluated achieved

the same level of control, suppressing the larval
populations to
high densities
nearly
zero
levels
in the
Cx. tarsalis was the only species encountered during the
as compared to the
during
course of this experiment.
the
Similar to the previous trials, the three Vectobac
The low densities of late instars present at 14 and 21
12AS rates of 0.2, 1. 0, and 2.0 kg/ha produced excellent

initial control three days after application yielding 99,
100, and 100 percent reductions, respectively. During
observed
check ponds
post- treatment period.
days post- treatment in both the treated ponds,
declining
the
result
of
populations
decreased
in the
as well as
this initial period, the late instar populations in the
check ponds, were the
oviposition
This
activity.
untreated ponds were still increasing, reaching a mean

density of 15 larvae per dip. By day seven, larvae
was
declining early instar counts in the

control ponds which by 14 days post- treatment averaged
0. 1 larvae per dip( egg rafts were absent), producing
by
confirmed
the
the rapid decline of late instars

never
became
and the
low
seen
irrespective of rate applied. As time progressed, the low
in Figure 4. Predation
factor due to the lateness
water
gradually began reappearing in the treatment ponds
larval populations showed only slight recovery,
of the season
remaining relatively unchanged for the rest of the one
temperatures, which suppressed their
month assessment period, which was very similar to the

trends seen in previous trials. A comparison of the post-
population growth.
Three
Cx.
and
DECEMBER, 1993
Cx.
species,
but because
of the
Cx. tarsalis,
treatment samples taken during the first week showed
in the third trial
that the larval densities in each of the treated ponds was
quinquefasciatus,
stigmatosoma were monitored
extremely low larval densities
Cx.
trials,
quinquefasciatus,
which
initially
was
dominant, eventually became replaced by Cx. tarsalis.

The shift in species composition of the ponds from
Cx. quinquefasciatus
result of a number of
depends
Cx. tarsalis
to
apparently the
Oviposition behavior
factors.
significantly lower than those in the control ponds
and
variability between ponds, a percentage

breakdown was not attempted. Again, as in the previous
considerable
was
ANOVA, P< 0.01).
However, the larval densities in
the control ponds declined steadily after three days posttreatment, and by day 14 the differences between the
treated and untreated populations had disappeared. All
pond densities were similar during the remaining two

weeks of assessment.
Other observations influencing mosquito densities
suitability of the oviposition site, and for

Cx. quinquefasciatus it is directly related to specific
in the fourth trial were important to consider. Early
water
ponds prior to treatment, continued to remain so
on the
conditions,
quality
organic
especially the
decomposition( Kaul
et al.
presence of
1977, Suleman
instar densities( 1st and 2nds), which were similar in all
and
throughout the assessment period, except for a temporary
oviposition
decline in the treated ponds three days post- treatment.
attractiveness of enriched pond water occurs shortly
This sudden suppression resulting from the B. t.i.
usually lasts a brief period of time

Mulla 1974, Mulla 1990, Beehler and
treatment was short-lived; and by day seven the densities
Shinn 1981).
after
This
flooding
Fanara
and
period of
heightened
and
Mulla 1993). Culex tarsalis is

conditions caused
by
strongly attracted to
abundant fermentation but rely on
not
generally associated with unpolluted habitats

Reisen and Meyer 1990). Once the relatively small
other cues
amount of supplemental organic material

ponds
to
had degraded, the
ponds
became
in
our
field
more attractive
ovipositing Cx. tarsalis adults.

A fourth trial was initiated in late March of 1992
the
of the young larvae had returned to their original level,

an indication that the effectiveness of the applications
was wearing off( data omitted).
In this trial, good oviposition activity was noted in

the check ponds throughout the course of the test,
eventually declining in the fourth week, based on both

egg raft numbers and early instar densities ( data not
included).
Because of the continuous oviposition
at
activity, one would expect the mosquito populations to
Coachella facility to substantiate the results obtained
eventually return to their original densities after the
earlier (
October 1991)
this site.
Because
the
treatments' effects had worn off,but no such resurgence
rapidly declining numbers of larvae in the control ponds

in the previous evaluation, it was difficult to tell how
was realized, as indicated in Figure 5. The inability of
much reduction of the
at
of
larval population in the treatment
ponds was due to natural phenomena and what proportion

could
be attributed to the efficacy of the Vectobac 12AS
applications( see
day
Fig. 3).
late instar populations to recover was due to the

abundance
of
predators.
Quickly establishing
themselves, various hydrophilid and dytiscid beetle
larvae were a common sight in dip samples three days

post-treatment. After two weeks, belostomatids and
Higher water temperatures prevailed during the 28-
dragonfly naiads had also become prevalent. Together
duration
the activity of these groups of predacious insects became
of this experiment
ranging from
a mean
131
DECEMBER, 1993
25
20
0-
0. 2 kg/ha
a-
1. 0 kg/ ha
2. 0 kg/ ha
a.
Check
15
A
4
1M
10
Z
00
g
Q
11.'.---
28
21
14
Efficacy of various rates of Vectobac 12AS against Culex larvae at Coachella Valley ponds( March-
Figure 5.
April 1992).
important factor in suppressing mosquito production

in the Coachella ponds as documented previously by
an
Walton
et al.(
The
and
M. S. Mulla.
1974.
Population
dynamics of Culextarsalis Coquillet and Culiseta
inornata ( Williston) as related to flooding and
1990).
results of
Fanara, D. M.
these studies,
under
the
conditions
temperature of ponds. Mosq. News 34: 98- 104.
reported here, demonstrated repeatedly that there is no

in using doses above the minimum effective
for a given habitat and situation) in order to extend
duration of control. Applications at minimum dosages
advantage
rates(
in conjunction
with
Kaul, H. N., B. L. Wattal, and P. Singh. 1977. Chemical

characteristics of Culexpipiensfatigans breeding
waters in areas around Delhi. J. Commun. Dis. 9:
8- 21.
the action of natural limiting factors,
appear to be sufficient to produce the desired extended

control of
Culex mosquitoes found in
ponds
filled
with
Lacey,
L. A.
and
C. M.
Lacey.
1990.
The medical
importance of riceland mosquitoes and their
nonpolluted water.
control using alternatives to chemical insecticides.

J. Am. Mosq. Contr. Assoc. Supp. 2: 1- 93.
REFERENCES CITED
Mulla, M. S. 1990. Activity, field efficacy and use of

Beehler, J. W.
and
oviposition
inhabiting
M. S. Mulla.
patterns
in
Temporal
Bacillus thuringiensis H- 14 against mosquitoes.
mosquitoes
Pp 134- 160 in H. de Barjac and D. J. Sutherland
1993.
Culex
managed marsh ecosystems.
Proc.
eds.),
Bacterial control of Mosquitoes and
Papers Calif. Mosq. Vector Contr. Assoc. 60: 85-
blackflies. Rutgers Univ. Press, New Brunswick,
S6.
NJ, 349 pp.
132
Mulla, M. S.
and
H. A. Darwazeh. 1985.
formulations
Efficacy
of
DECEMBER, 1993
Reisen, W. K. and R. P. Meyer. 1990. Attractiveness of
of
Bacillus thuringiensis H- 14
selected oviposition substrates for gravid Culex
against mosquito
larvae. Bull. Soc. Vector Ecol.
tarsalis and Culexquinquefasciatus in California.
10: 14- 19.
J. Am. Mosq. Contr. Assoc. 6: 244-250.
Mulla, M. S., B. A. Federici, and H. A. Darwazeh. 1982.

Larvicidal activity of Bacillus thuringiensis
serotype H-
14 against stagnant- water mosquitoes
Suleman, M. and M. Shirin. 1981. Laboratory studies

on oviposition behavior ofCulexquinquefasciatus
Say( Diptera: Culicidae): Choice of oviposition
Environ.
medium and oviposition cycle. Bull. Ent. Res.

71: 361- 369.
Mulla, M. S., R. L. Norland, D. M. Fanara, H. A.

Darwazeh, andD. W. McKean. 1971. Control of
Walton, W. E., N. S. Tietze, and M. S. Mulla. 1990.

Ecology of Culex tarsalis( Diptera: Culicidae):
factors influencing larval abundance in
in recreational lakes. J. Econ.
mesocosms in southern California. J. Med. Ent.
and
its
effects on nontarget organisms.
Ent. 11: 788- 795.
chironomid midges
Ent. 64: 300- 307.
27: 57- 67.
DECEMBER, 1993
THE ORIENTAL LATRINE FLY, CHRYSOMYA MEGACEPHALA

FABRICIUS 1794) ( DIPTERA: CALLIPHORIDAE),
AS AN INVADING BLOW FLY OF PUBLIC HEALTH IMPORTANCE

A. R. Olsenl, T. H. Sidebottom2, and S. G. Bennett3
ABSTRACT: The Oriental latrine fly, Chrysomya megacephala( Fabricius)( Diptera: Calliphoridae), is a blow fly
pest of public health importance that has invaded many new localities over the past two decades. It is a carrier of
numerous food-borne pathogens, including Shigella and Salmonella, as well as infective stages of intestinal
parasites. The distribution of this blow fly has expanded from its original range in the Orient and Australasia to
localities in Africa, South America and, most recently, North America. Its biology, systematics and public health
importance are reviewed for vector control and public health entomologists in the localities that it has recently
invaded. Monitoring the spread of C. megacephala is necessary to minimize its impact on sanitation and health in
the new territories that it invades. Food manufacturing and preparation establishments are at special risk from
increased insanitation attributable to the Oriental latrine fly and must practice good sanitation in order to prevent the
contamination of foods by this pest.

SYSTEMATICS AND MORPHOLOGY
INTRODUCTION
The Oriental latrine
fly
that has recently
World
throughout the
Within the
fly
1988)
taming
regions
of the earth.
the Indian bazaar fly. The species type was designated
has invaded
by Coquillett 1910 from Chrysomya regalis Robineau-
and
it
North America,
and
requires
and
its
of the
invading
with
literature( Zumpt 1965).
species.
The
blow
fly
is
personal communication).
is known
about this
monitoring its
encourage additional
new environments.
Dear 1985a) as a facultative parasite of wounds
always available
Kitching 1976).
invader( J. C.
Egg; white, 1. 4- 1. 5 mm. Chorion smooth, without
review of what
sculpturing. Median strip narrow, running almost the

full length of the egg. Micropylar pit smooth, without
chitinous fringing( Kitching 1976).
study of this insect as it

This review covers the
to
Mature larva( after Erzinclioglu 1990); white, 14 to
published
17 mm long. Cephalopharyngeal skeleton with mouth-
spreads
Oriental latrine fly and contains
unpublished
observations
for Southern California.
1U. S. Food&
fly
is necessary in order to continue

in the U. S. and other areas, and to
studies and reports of the
Chrysomya bezziana, the Old
filth fly that is only occasionally involved in myiasis
reports con-
fly
spread
A full
are
a recent
Mariluis,
Peris 1984).
World screw- worm fly, is an obligate agent of myiasis

of livestock ( Ferrar 1987) while C. megacephala is a
systematics and
the
and
and Chrysomya bezziana Villeneuve 1914 were often

assigned to Chrysomya dux( Eschscholtz 1822) in the
Effective monitoring of an
entomologists and public health
to be familiar
Mariluis
taxonomic synonymy is provided by Zumpt who also

notes that up until 1914 both Chrysomya megacephala
Greenberg
in the Nearctic
range
in regional journals that are not
previously
Desvoidy 1830 (
has
documenting the movement
expands
areas where this
records
fly
this information for the Oriental latrine
scattered
in the
Calliphoridae), the Oriental latrine fly, is also known as
other regions.
professionals
bionomics
from the Old
warmer
cited the necessity of
invasion
Chrysomya megacephala( Fabricius 1794)( Diptera:
range
Baumgartner( 1988)
of this species as
Region and
disease- carrying blow

its
in Southern California in three
established
coastal counties.
decades, this filth
past two
Africa, South America,

become
is
expanded
and
collection
hook tooth much longer than depth of base; oral sclerite

small and pigmented at base, unpigmented at the tapering
anterior end; dental sclerite weak; liguloid arch narrow
Drug Administration, 50 United Nations Plaza Federal Office Bldg., Rm. 526, San Francisco, CA
94102, USA.
2U. S. Food&
Drug Administration, Division of Field Science, Room 12- 39, 5600 Fishers Lane, Rockville, MD
20857, USA.
Grove, CA 92643, USA.
3Orange County Vector Control District, 13001 Garden Grove Blvd., Garden
134
but well- pigmented;
windows
and ventral cornua; angle
Anterior
cornua wide.
processes.
on
2- 8,
on
9,
spiracle with
spinal
dorsal
lower third of the eye. Females dichoptic with eyes

separated, upper facets not enlarged. The male genitalia
10- 13 fingerlike
are illustrated by various authors ( Hardy 1981, Prins
bands ( segments 2- 12)
1982, Kurahashi 1991) as are the genitalia of both sexes
complete
laterally but present dorsally and ventrally

incomplete dorsally on 10- 12; bands on 6- 12 cleft
absent
ventrally; bands on 2- 8
Posterior spinal bands (
dorsally
Spines
than
on
and ventral
Posterior spiracle with unpigmentedecdysial
Anterior
scar.
usually present
between dorsal
on
6- 10,
6- 11)
Identification of adults to the genus level can be
band,
anterior pleural
subfamily Chrysomyinae ( Peris 1992).
incomplete
dorsally
Papilla P2
ventrally. Most spines with two or

1991, Meng et al. 1966).
and
analytical key ( after Kurahashi 1991) may be used to
Holloway
closer to
P1 than to P3.
Body
lacks
differentiate C. megacephala adults from other

Chrysomya species that are potential invaders into the
fleshy
1- 11.
projections or papilla on segments
The genus is
defined by a broadly triangular thoracic squama with

long dorsal hairs combined with the presence of dorsal
setulae on the stem vein of the wing. The following
11 but faint
ventrally.
but fewer laterally
complete on
Kano and Shinoga 1965, Thomas 1951).

accomplished using a key to world genera of the
present all round anal region
three teeth each (
lateral
with
segments
DECEMBER, 1993
An
United States.
earlier description of the larva ( Patton and Cushing

1934)
does not include key characters of the
cephalopharyngeal skeleton.
The
puparium
spiracles and
which
are
by
may be
recognized
by
Other geographic areas will require keys that include

all the indigenous species of the area. Keys are available
for identification of Chrysomya species from the Old
World ( Zumpt 1965, Senior-White et al. 1940), Asia
Kurahashi 1967, Singh et al. 1979, Tumrasvin et al.
the posterior
the patterns of the bands of spines
retained
from the larval
stage (
Kitching
1976).
The
adult
is
blow
a robust
fly,
8 to 12
body
1979), South America ( Mariluis 1981a) ( in Spanish)
blue. Frons black with the lower

part of the head( buccae) orange. Mesonotum with
two
short longitudinal vittae or dark stripes
anteriorly.
Anterior spiracle dark brown to black with a black
and Australasia( James 1971, Kitching 1976). Keys for
mm;
color metallic greenish
pre spiracular spine( stigmatic
Abdominal
bristle)( Kurahashi 1991).
segments black- banded on posterior margins.
Wing hyaline with the base of the radial vein ciliated

dorsally before the humeral crossvein.
identifying adults and larvae of the major public health

(
pest species of every zoogeographical area are available
Greenberg 1971). Finally, forensic keys often include
Chrysomya megacephala and other Chrysomya spp.

found on carrion and cadavers( Smith 1986).
Lower( thoracic)
DISTRIBUTION
squama large, triangular, brownish, white basally, haired

above with
Male
long
portion and a
1.
2.
setulae.
eye with upper
Legs black. Male holoptic.
facets larger
distinct line
of
than
facets
demarcation
Prothoracic
spiracle white
Prothoracic
spiracle
on
lower
across
the
There is speculation concerning the origins and

dispersal routes of C. megacephala. The origin of C.
megacephala was once placed in Papua New Guinea
dark
Legs
metallic black;
parafacials( buccae) yellow to orange, at least anteriorly; wings hyaline
in both
front femur
sexes;
of male with no conspicuous white
hairs
Legs testaceous yellow in part; parafacials orange, densely yellow dusted; Front femur of
male with
3.
4.
dense
erect white
Prespiracular
seta present
Prespiracular
seta absent
hairs
dorsally
Chrysomya varipes( Macquart)

Chrysomya rufifacies( Macquart)
Chrysomya albiceps( Wiedemann)
Body stout, but rather elongate, submetallic dark blue; thoracic squama largely blackish
Chrysomya pacifica Kurahashi
Body stout and rounded, metallic greenish blue; thoracic squama largely brownish
Chrysomya
megacephala(
Fabricius)
DECEMBER, 1993
based on the
finding of a"
form there
natural"
and
in the
Pacific islands( Kurahashi 1982, Mihara and Kurahashi

1991). It was later determined that the supposed"
form
was a separate
Kurahashi 1991)
is
from C.
species
and the true origin of
natural"
C. megacephala
recent Nearctic records discussed below.
The first confirmed North American record of C.
The Oriental latrine fly is indigenous to the Oriental
megacephala was from peninsular Baja California,
Australasian Regions ( Bezzi 1927, Dear 1985b,
Mexico(
James 1971),
Greenberg
Park
and penetrates the
peninsula(
Palearctic in Afghanistan
1971), Japan( Kurahashi 1967),
Kano 1963).
and
warmer areas of the
It is found
Indian Ocean
on
Hawaii(
1940)
and
a fishing port barely 150 km from the United States

border( Bennett 1989). Both Mexican localities are on
Malaysian
the Gulf of California where it is likely that fishing and
far
as
east
Samoa( Malloch 1930),
other maritime vessels
earlier (
1960)
inclusions
of these two countries
range except
specimen
Prior to 1988, the genus Chrysomya Robineau-
one record of a single
adult male
flies, and Cole ( 1969) reported that the genus did not
1988).
Oriental latrine
by
fly
Shewell' s ( 1987)
also
rufifacies( Macquart), introduced to Texas and Oklahoma

(
Baumgartner
and
Greenberg
1984).
A single male
Thecontemporary patternofdispersal
trap along the Texas- Mexico border( Wells 1991) but
of this
first
this record is apparently similar to the earlier interception
an area
recorded from Puerto Rico and not from an established
fly is established( Kurahashi 1978, Baez et al.

and the fly then spreads inland
population. Although isolated specimens of Chrysomya
a new
territory
consists of
handle trade from
Prins 1982). In late 1977, it
1978, C.
North America.
occurrence ofa related screw- worm species, Chrysomya
to other
modern expansions of
1981, Gagne 1981)
continent
western
specimen of C. megacephala was recorded from a wind
records near seaports that
the
in
fly ( Gagne 1981,
dispersal
C. megacephala into
where
occur
treatment of the Nearctic blow flies mentions only the
Baez 1990). Air transportation
contributes to the
Greenberg
1961
1984).
Greenberg
of the
trade (
Hall ( 1948) does not include the modern
Chrysomya flies in his manual of North American blow
and
spread
1955).
Prior to
range.
outside this
regions was made possible

maritime
Desvoidy 1830 sensu Coquillett 1910 was notknown to

occur in California.
It was not found during a
comprehensive survey of California blow flies( James
known from
was not
intercepted in Puerto Rico in
Baumgartner
The
for
in the
transportation
Mexico.
also
but Kurahashi ( 1978), working with an

version of Park' s study, questions the
1977, C. megacephala
will provide
opportunities for the fly along the entire west coast of
as
occurring in Fiji( Kurahashi 1981). Park( 1977) includes

Egypt and Iran within the natural range of C.
megacephala
Baumgartner 1988). It was
Korea
and
Kurahashi 1979, Reid 1953). It is generally
Hardy
Greenberg 1988,
later found on the Mexican mainland in Puerto Penasco,
islands in the
and on the
distributed in Oceania ( James 1947)
of
reported no C. megacephala from that country

Greenberg and Szyzka 1984, Baumgartner and
Greenberg 1985). TABLE 1 summarizes the known
distribution of C. megacephala as of 1993, including the
megacephala,
yet unknown.
and
135
was
found
on
spp. have been collected in the New World since the last
the African
century ( Baumgartner
in Ghana and in Senegal( Kurahashi 1978). In

megacephala
Africa( Prins 1979)
was
where
found in
it is now
localities( Braack 1991). The
Greenberg
1984),
those
South
and prior to the late 1980' s in the Nearctic do not
in inland
represent established populations ( Baumgartner and
coastal
widespread
and
collected prior to the 1970' s in the Neotropical Region
it was found in
Greenberg 1985, Wells and Greenberg 1992).
Canary Islands( Baez et al. 1981). At about the same

time, the fly was found in Brazil but the actual invasion
Examinations of repository collections at California

Academy of Sciences ( CAS), Los Angeles County
Museum of Natural History( LACMNH), U.S. National
Museum of Natural History ( NMNH) ( Smithsonian
Institution), and San Diego Museum of Natural History
same year
the
of
Brazil
occurred earlier (
do Prado
and
GuimarAes
1982). Brazilian dipterists postulate that C. megacephala

was transported there
refugees
from Angola
during
in 1975- 1976 ( GuimarAes
an exodus of
et al.
1979, Peris
1987),
predating the first published records from

subsaharan Africa. In South America, it subsequently
spread
and
to Argentina( Mariluis 1981 b),
Paraguay(
Barrios
Peris 1983), Venezuela( Baumgartner 1988),
Ecuador( Olsen,
and
1992). The Ecuador invasion
SDMNH) found no specimens of C. megacephala

collected in California prior to 1988. The NMNH has
specimens of C. megacephala collected in Fort
Lauderdale, Florida in June, 1990.

The first established population of C. megacephala
by
in the continental United States was discovered on
C. megacephala is apparently recent as it was not noted

from collections madeinthisareain 1978- 1980( Mariluis
August 6, 1988 at Scholl Canyon, a landfill site in the
1981b).
et al.
The Oriental latrine
Peru( Laurence 1981) but
fly
was said
subsequent
blow
foothills of the San Gabriel mountains north of
to occur in
metropolitan Los Angeles ( CDFA 1988). The Scholl
fly
Canyon record is from
surveys
emergence traps that were
being
136
DECEMBER, 1993
TABLE 1. Current distribution of Chrysomya megacephala by zoogeographic regions.
ORIENTAL
OCEANIA
AUSTRALIAN
PALEARCTIC ETHIOPIAN NEOTROPICAL NEARCTIC
China
Australia
Carolines
Afghanistan
Angola?
India
N. Zealand
Fiji
Iran?
Canary
Indonesia
Papua/ N.G.
Gilberts
Japan
Egypt?
Ecuador
Malaysia
Guam
Korea
Ghana
Paraguay
Philippines
Hawaii
Malagasy
Peru?
Sri Lanka
Marianas
Mauritius
Venezuela
Taiwan
Marshalls
Senegal
Thailand
Palau
Seychelles
Viet Nam
Samoa
South Africa
I.
Argentina
Mexico
Brazil
U.S. A.
Solomons
Tonga
Vanuatu
indicates that
latrine
few
fly activity ( Poorbaugh 1989) and

breeding population of the Oriental
already established by 1988. Within a
to monitor
used
fly
was
weeks, numerous adult males and
collected
1990),
in
and
Los Angeles( Olsen
urban
C.
Diego were collected at a commercial fish packing

establishment that receives large amounts of product by
megacephala
Southern California
has
since
females
and
been
truck from the Los Angeles area. It is likely that the San
Diego flies were transported south from Los Angeles in
were
Sidebottom
view of their absence from the nearby border
in
communities. Voucher specimens of the authors' recent
collected
Altadena,
collections in California are deposited at the San Diego
Anaheim, Brea, Burbank, Cypress, Fullerton, Garden
Museum ofNatural History and the California Academy
Grove, Huntington Beach,
of Sciences.
the
communities
Long
of
Beach, Orange,
Pasadena, San Diego, and San Marcos. These collection

localities
bring
California
three coastal
Angeles, Orange,
counties (
Orange
The
immatures
counties
year
likely
have
examination
dumpsters)
Ysidro
and
of
20
was made
Otay
the fly would find the intervening desert a formidable
in the U.S. is
and air traffic from those localities is very limited. The

apparent absence of C. megacephala from likely border
have
fly
occurred
in August,
crossings confirms the unlikelihood of the California
the border with
sites ( meat
colonization
originating from
western
Mexico.
An
alternate route into California is by air transport from

Hawaii, a recognized staging area for insect invasions
market
July and August of 1991
in San
into California ( Wilton 1961). Hawaii is within the
border crossing
of San Diego. No adult or larval
Hawaii to California invasion route is not unreasonable
Mesa, the
communities just south
may have
Mexico, an
megacephala
attractive
in
An overland
barrier to overland migration into the U.S. via California,
or shortly before.
To determine whether C.
across
uncertain.
and Gill 1989), but from the known localities in Mexico,
1988
invaded California
are
from 1988 through 1993.
permanent establishment of this
to
authors
in California
migration from Mexico has been postulated ( Dowell
in Los Angeles and
and adults
every
confirmed and most
arrived
San Diego) into the known
and
distribution range of C. megacephala. The

collected
The route or routes by which C. megacephala
Los
traditional range of the species ( James 1947) and the
principal
C. megacephala were found at these localities along the

California- Mexico border. All specimens from San
Baumgartner 1988). Near the Scholl Canyon site there

an international airport at Burbank, California, that
is
DECEMBER, 1993
daily flights from Hawaii, and

recorded locality for C. megacephala.
receives
Pacific invasion
mutes that
include
megacephala
have been
Shanghai
Burbank is a
Other
for C.
Honolulu
route
to
Esser 1991a). The post- emergence sex ratios of adults

captured in the wild are biased towards females. This
pan-
postulated
was first noted in India and attributed to habitat selection

(
Illingworth 1926) and one from Hong

E. Easton,
personal
Kong to California
communication), both aided by
where they were more easily captured by researchers.
shipping and transportation. An analysis of the

karotypes of Chrysomya spp. from Brazil indicated that
megacephala
Espin
traveled there from Japan ( Azeredo-
Pavan 1983).
These
Other studies attribute sex bias to a differential attraction
of females to baits used in traps, either for feeding

(
Avancini 1986, Mariluis
and
Schnack 1985)
or
the chief
oviposition( Spradbery 1979, Das and Dasgupta 1982b).

Females are differentially attracted to various substances
depending on the stageof ovarian development( Avancini
to accepting a migration route across the
and Linhares 1988). Sex bias due to differential responses
and
reports
postulate of a
Pacific invasion
There is
for further study, but for
a need
objection
Roy and Dasgupta 1980). Males were thought to prefer

shaded habitats while females occurred in open habitats
modern
C.
137
Pacific to California for C.

invasion did
not occur
support
for California.
route
now,
megacephala
decades
the
is why the
to trap baits between the sexes is a common occurrence

in blow flies ( Muirhead-Thomson 1968).
ago.
Another
possibility is that differential susceptibility to the fly

BIOLOGY
pathogen Staphylococcus muscae may cause early dieoff of males as soon as 15 days after emergence( Mariluis
The life history
of
C. megacephala is described by
Wijesundara( 1957a, 1957b). The life
cycle
Schnack 1985, 1989).
and
from egg to
Whatever the reason, the
observed larger proportion of C. megacephala females
adult is
is statistically significant ( Roy and Dasgupta 1975,
or
approximately 8 to 10 days at room temperature,

6, 415 accumulated degree hours ( Goff 1992), but
Mariluis et al. 1990).
may take up to 13 days( Prins 1982). The egg stage is

9 to 10 hours duration in most cases but may last up to
The newly-emerged female reaches sexual maturity

in one to three weeks ( Esser 1991a) although mating
15 hours before hatching( Esser 199la). The larval stage
activity may begin on the third day after emergence.

The female is capable of ovipositing 150 to 392 eggs
during her life span. A single female may oviposit up to
consists
three instars and the larval period lasts
of
days ( Prins 1982).
two to six
nonfeeding
to pupate( Bohart
order
and
Mature larvae
from the
stage and migrate
feeding
Gressitt 1951).
enter a
site
The
100 eggs in a three minute period ( Subraimanian and
in
Mohanan
non-
1980b).
The usual breeding medium is
lasts up to one day at room

feeding
temperature. The duration of the second and third instar
excrement( Norris 1965, Goff et al. 1991) but females
larval
that are suitable for larval development. These include
prepupal
prolonged
numbers of
while
on
ratio
days to
complete
Goff 1990). This
will feed and oviposit on a wide variety ofother substrates
larval
to
density and
density. Large
per weight unit of available
periods of
larvae under low density
over seven
and
larvae
in inverse
in larval developmental
result
less,
is partly dependent
stages
may be
stage
five days
conditions
garbage( Kano 1958), carcasses( Goff et al. 1986, Early
food
seafoods( Olsen and Sidebottom 1990), fresh red meat
or
may take
development( Goodbrod
appears
to be
body
size and weight at pupation
along
food
larvae that do
source
Greenberg
The
is
exhausted
do
before
not survive to adulthood
1991).
pupal stage
longevity
not reach this plateau
for both
is usually five to six days. Adult

sexes is 47 to 90 days at room
temperature. The total egg- to- egg life cycle is 18 to 19
days,( Subramanian
to
20
and
Mohanan 1980b), allowing up
generations per year under optimal conditions.
In
adults
laboratory
rearings, the sex ratio of emergent
is nearly 1: 1( Subramanian
and
Mohanan 1980b,
Dasgupta 1982a), chicken meat ( Linhares
and cadavers(
Goff
et al.
the same site ( Esser 1990).
with
increased mortality. Beyond the plateau of 45 percent

of average body size, larvae can develop into viable
adults while
and
1991). Dried, cured and
salted fish is acceptable for breeding( Haines and Rees

1989). Group oviposition is common, with the eggs of
one female attracting additional females to oviposit on
strategy
larvae from a
limited food resource. The fastermaturing larvae exhibit

reduced
Das
1981)
a survival
to produce the maximum number of
Goff 1986), foods ( Bohart and Gressitt 1951),
and
Females are known to
oviposit on open wounds and sores of animals where the
larvae become
facultative
agents of myiasis
( Subramanian and Mohanan 1980a). RoyandDasgupta

(
1971) demonstrated that larvae will produce myiasis
on unbroken skin of living animals if the eggs are placed

on the skin artificially, but females cannot be induced to
oviposit on unbroken skin even though they will lay
eggs on open wounds and sores. The Oriental latrine fly
is therefore primarily saprophagous because of female
preferences for oviposition sites.
The Oriental latrine fly is heliophilic ( Linhares

1981,
Mariluis
and
Schnack
1985),
exophilous
138
1971)
Greenberg
and
synanthropicinbehavior.
fly,
it is
1951,
attracted
Roy
and
1990). It is
indoors to food( Bohan
Dasgupta 1975, Olsen
also attracted to
excrement ( Jettmar
both indoors
summarizes
been
and
where
1940, Kano 1958)
Gressitt
Sidebottom
( Schnack
it breeds in
et all.
1989). Negative values indicate an
inability to survive in close association with human
and to corpses
Goff 1991).
SI) for blow flies( Nourteva 1963) where, on a scale of
100 to + 100, a calculated value of zero is neutral
settlements
and
positive
values
reflect
progressive
degrees of synanthropy. A survey in India using baited
the attractive substances and sites that have
traps determined the SI of C. megacephala to be+ 91. 6
and outdoors (
in the literature for C.
The degree
different
latrines
and
developed to calculate a relative index of synanthropy

(
TABLE 2
reported
in human
to hemi-
eusynanthropic
Although basically an outdoors
DECEMBER, 1993
of
synanthropy,
settlements, varies
geographical
areas.
or success at
for C.
Roy and Dasgupta 1980), indicating the fly was adapted
megacephala.
megacephala
measure
to survival in or near human settlements.
surviving
in
has been
Similar
surveys found SI values ranging from+ 75. 2 in Brazil

(
Ferreira 1978, Linhares 1981) to+ 32. 8 in Argentina
TABLE 2. Observations of attractive sites for adult Chrysomya megacephala.
ATTRACTION
LOCALI'1' 1ES OBSERVED
Foodstuffs
general
Brazil, Guam, Malaysia
eggs
U.S. A.
fish
Brazil, India, Malaysia, Palau, S. Africa, Sri Lanka, U.S. A., Venezuela
fruit
China, Fiji, Guam, India, U. S. A.
meat
Brazil, Guam, Hawaii, Japan, Malaysia, Sri Lanka, U.S. A.
melons
China
shellfish
Guam, Solomon I., U.S. A.
sweets
China, Guam, Sri Lanka
Structures
houses
China, Guam, Japan, Palau
markets
China, Ghana, Malaysia, Senegal, Sri Lanka, U.S. A.
latrines/ toilets
Guam, India, Malaysia, Palau
restaurants
Mexico, U.S. A.
ships
S. Africa, mid- Pacific
slaughterhouses
Sri Lanka
Other
carrion
Australia, Brazil, Guam, Hawaii, Malaysia, Papua/ New Guinea, S. Africa, Sri Lanka
corpses
Guam, Hawaii, New Zealand, Sri Lanka
feces
China, Brazil, Guam, S. Africa, U.S. A.
garbage cans
China, Hawaii, Malaysia, U. S. A.
landfills
Malaysia,
Paraguay, U. S. A.
Mariluis et
less
with
of
al.
1990). These calculations
mathematical observations of
C. megacephala for
been reported for C. megacephala. Cluster swarms of
are consistent
adults occur in Okinawa under the boughs of shade trees
the preference
habitats from
urban and village
( Bohart
1991) to
apartment
complexes
in Hawaii
Gressitt 1951).
( Bennett 1989).
Similar swarms have been
In the Pacific islands aerial nuptial
swarms of males have been described ( Olsen and
Wilton 1961).
we
and
seen in California, numbering thousands of individuals
Guimaraes 1982) to New Zealand
Brazil( do Prado&
Holloway
139
DECEMBER, 1993
In the temperate climate of Los Angeles, California,
Sidebottom 1990). Fly species with sexually dimorphic
larval activity
holoptic males, such as C. megacephala, often exhibit
have observed a cessation
during
indicate that the
pupal stage
Chrysomya
species,
in
puparia
temperatures as
of cooler
C.
areas,
Das
is
prolonged
is the
In
with
Chrysomya megacephala can compete with and
This may
displace indigenous flies of public health importance
during periods
case with a related
Wiedemann), in
central
were infested with C. megacephala contained few house
bimodal
flies, Musca domestica L. (Muscidae), or greenbottle
per
flies, Phaenicia sericata( Meigen)( Calliphoridae). After
exhibit
two
population peaks
the containers were cleaned and C. megacephala
1982a). These fluctuations may

to the availability of suitable food and
and Dasgupta
response
Baumgartner and Greenberg 1984). In some Orange

County collection sites, for example, dumpsters that
tropical and subtropical
populations
megacephala
fluctuations
seasonal
protected sites.
albiceps(
Asia( Sychevskaya 1981).
be in
this type of aerial nuptial swarming( Oldroyd 1964).
the coolermonths from November through March
but have found
year(
of adult and
removed, the other species returned to the sites.
oviposition sites. Temperate climates result in a unimodal

fluctuation regulated by temperature(
FOOD SANITATION
Roy and Dasgupta
1975).
There
are
natural
of
a number
enemies
The association of the Oriental latrine fly with the

transmission of food- borne disease is firmly established.
of the
Oriental latrine fly. Ants, especially Solenopsis geminata
In 1938, an outbreak of dysentery in China was linked

to a population peak of this fly that occurred in the
F.) and Pheidole megacephala( F.), are majorpredators

of the
larvae( Tullis
McCook
was
pupae near a
and
Goff 1987). Solenopsis xyloni

and
absence of M. domestica and other disease carriers
California.
( Jettmar 1940). At the same time adult Oriental latrine
predaceous
histerid beetles
flies in China were found to harbor and transmit bacteria
flesh
larvae ( Diptera:
carrying
dumpster in Orange
observed
The larvae fall prey to
Coleoptera: Histeridae)
and
off
live larvae
County,
fly
Sarcophagidae) that hunt prey in the offal and carrion

The larva of a
where the Oriental latrine fly breeds.
closely related species, Chrysomya rufifacies, may prey
on C. megacephala and other blow fly maggots under
crowded conditions(
cited
previously
life
Wells
et al. 1988b, Sulaiman et al. 1989), including infective

ova ofAscaris lumbricoides and Trichuris sp. Pathogens
and parasites including the above as well as Giardia and
the waspNasoniabrevicornis( Ashmead)(
Pteromalidae) ( Hardy 1981). A
al.
1981).
We
and other
observed predation
C. megacephala in Los Angeles

where
both
species are
Pacific island
spider,
Vespula
of
Nephila
by
V.
ones.
fly
longevity ( Ho 1990). Vertebrate
phoretic
on
adults where excessive numbers
feed opportunistically
1981),
On the
Fabricius) ( Araneae:
on adults(
Public health
Cushing 1934), transmitter ofintestinal pathogens( Hardy
Orange Counties,
Scopoli) ( Acarina: Macrochelidae) is
reduce
Down 1946).
contaminator of foods ( Illingworth 1926, Patton and
Tetragnathidae), capturing numerous adults in its large

aerial webs. The mite Macrocheles muscaedomesticae
Oriental latrine
and
et
we observed the giant wood
maculata (
Harris
entomologists characterize this species as an important
germanica on
recently introduced
Palau,
megacephala(
preys
blow flies( Akre
and
tapeworms have been isolated from feces of C.
Hymenoptera:
yellowjacket,
Fabricius)( Hymenoptera: Vespidae),
C. megacephala
et al. 1988a) and a variety of other food-borne pathogens

Greenberg 1973). Intestinal parasites are transmitted
mechanically by this fly( Monzon et al. 1991, Sulaiman
among C. megacephala larvae, even under

dense crowding. The pupa and larva are parasitized by
on adult
Subsequent studies have linked C.
1992). In the
Greenberg
not observed
germanica(
Chow 1940).
megacephala with enteropathogenic bacteria( Sulaiman
studies, cannibalism was
and
history
responsible for dysentery and other enteric diseases

(
may
insectivores
Gressitt 1954).
There are two types of swarming behavior that have
and
disease ( Kurahashi
and
Banu 1989),
and
more dangerous than the house fly in these respects

(
Bohart and Gressitt 1951, Greenberg 1988, Monzon et

al. 1991).
In order to avoid the transmission of food- borne

pathogens and human parasites, food manufacturers
must use a HACCP( Hazard Analysis Critical Control

Point) system to prevent contamination from these
organisms(
Gorham 1989). Critical control points for
this purpose include testing of raw materials, biocidal

steps
that kill microorganisms and exclusion from food
140
or food contact surfaces ofpotential carriers ofpathogens
Because
Oriental latrine fly is more

dangerous than house flies as a carrier of disease
Greenberg 1988), the presence of this fly in a food
and parasites.
handling
the
increases the probability that
area
be transported from
will
very dangerous to
places
pathogens
they may
places where
not
be
their potential for
where
becoming hazard is greatly amplified. The potential

for transmission of food- borne pathogens
by C.
a
megacephala
Bohan
and
is considerable for
a number of reasons
Gressitt 1951). It readily
by
when attracted
various
food
buildings
enters
products and
is
also
DECEMBER, 1993
legal action by authorities even though actual

transmission of a specific pathogen at the time of
observation
is
not
demonstrated.
This is because
unacceptable increases in the risk that transmission of
pathogens will occur as long as the flies are allowed to

freely enter andexit places where food is handled.
Even in the absence of direct observation, evidence
that a food was not protected from fly activity may be

detected. There are a number of analytical techniques
for detecting cryptic insect filth in food products( Boese

and
Bandler 1990).
These " light filth"
extraction
techniques use principles of physical chemistry to
attracted to reservoirs of pathogens and other
separate insects and insect fragments from food products.
items that
unsavory
environments where it
Once extracted, the insects and insect fragments are
it easily becomes
transmitting its full
micromorphology of adult fragments of C. megacephala
As
thrives.
in
mired
occur
in the
a rather
food
urban
clumsy
bacterial load in the
animal,
dies there,
and
The TABLE 2 list
process.
observations of the attraction of C. megacephala to

products, structures and pathogen reservoirs
its
to
potential
for
transmitting
pathogens
from
food products. The HACCP objective is to
megacephala
from facilities in
of
food
illustrates
identified
to
determine their etiology.
The
provides taxonomic information that is sufficient to
identify
these
fragments ( Kano
and
Sato 1951).
The
presence of microscopic insect filth in food is
reservoirs
objectionable even without a direct link to pathogenicity
C.
but specific identification of insect filth, from blow flies
the
or other insects, is an important step towards evaluating
exclude
order to prevent
transmission of pathogens from these unsavory types of
the relative risk to consumers( Kurtz and Harris 1961).
reservoirs to food or food contact surfaces, especially at

those points after the last critical biocidal
step in a food
CONCLUSION
handling process( Gorham 1991).

Avancini
1992)
and
Ueta ( 1990)
megacephala to reduce
from
and
evaluated several pesticides
et
control of
al.
There are ample reasons for concern over the spread
C.
of the Oriental latrine fly across the warmer regions of

the world. Not only is this blow fly a major public health
losses in the poultry
avian parasites carried
by
industry
fly. Studies of
the food industry
the
Chrysomya blow fly control in

demonstrated moderate success for
but there are
Sulaiman
for the
certain pesticides,
serious concerns over the pesticide residues
that remain after treatment( Walker and Donegan
1988).
A concern with any chemical control is the

ability of live
C. megacephala larvae to retain pesticide residues in
their tissue(
Gunatilake and Goff 1989). For the present,
it is necessary
to
measures to protect
rely
on prevention
foods from
and exclusion
contamination
by these
flies. Remedies thatdo not requirepesticide applications

on or near
control(
foods
are the optimal
long- term
solution to
Esser 1991b).
adults and their
fragments
were
identified from
imported frozen shrimp and other seafoods by the Los

Angeles laboratory of the U. S. Food and Drug
Administration( FDA)( Olsen et al. 1992). In 1989, a
Los Angeles market was cited for health code violations
by local
authorities,
among them
being
the presence of
blow flies inside the market on exposed fresh produce,

meat and
baked
the potential to successfully compete with and displace

indigenous blow flies in urban settings. Public health
entomologists and other officials must be aware of the
habits, appearance, and biology of this pest species,

particularly in geographic localities where the Oriental
latrine fly is recently introduced or localities that are

vulnerable
to invasion
by
this filth
fly.
Adequate
preventive and control measures are only possible if the

pest is recognized and its potential impact understood.
The Oriental latrine fly is not subject to quarantine,

eradication, or other statutory and administrative barriers
that apply to its close relatives, the screw- worm flies.
Long before C. megacephala arrived in California,

dead
pest in terms of transmission of disease, but it also has
goods.
Over 25 live
adult
flies, mostly
Once established in a locality, local public health officials

have little recourse but to monitor its spread and
encourage measures that will limit the availability of

breeding sites, exclude the species from buildings where
foods are being processed, and remove potential
reservoirs of pathogens that could be visited by this
disease- carrying pest.

Acknowledgments
C. megacephala females, were observed inside the

market.
Observations
such as
these lead to corrective
Thanks to James J. Madenjian, Daniel F. Gross,
DECEMBER, 1993
Nicholas J. Richter
and
J. Richard Gorham(
David Faulkner ( SDMNH)
and
141
FDA),
Baez, M.,G. Ortega and H. Kurahashi. 1981. Inunigration
R. L. Berstein, San
of the Oriental Latrine Fly, Chrysomyamegacephala

Fabricius) and the Afrotropical Filth Fly, Ch.
chloropyga( Wiedemann), into the Canary Islands
all
Francisco State
University, for technical comments

and to Johnson Toribiong and Angelina Smaserui Olsen,
Kerallang Natural History Institute ( Palau), for
Diptera, Calliphoridae). Kontyu, 49: 712- 714.
specimens and background information on C.

megacephala
in the tropics.
We
also thank
Juan C.
Mariluis, Instituto Nacional de Microbiologia" Carlos
Barrios, B. B. and S. V. Peris. 1983. Chrysomya mega-
cephala( Fabr., 1784) en Paraguay. Eos 59: 17.
G. Malbran"( Argentina); Juan A. Schnack, Universidad

Nacional de La Plata( Argentina); Hiromu Kurahashi,
Dept.
of
Medical
National Institute
Entomology,
Health ( Tokyo, Japan); Emmett Easton,
University
IL); Gilbert L. Challet
and the
1988.
Spread of introduced
Chrysomya blowflies ( Diptera, Calliphoridae) in
of
the Neotropics with records new to Venezuela.
East Asia, visiting professor ( Macau); Donald L.

Baumgartner, Northwest Mosquito Abatement District
Wheeling,
Baumgartner, D. L.
of
Operations
Biotropica 20: 167- 168.
Baumgartner, D. L. and B. Greenberg. 1984. The genus
Staff of Orange County Vector Control District; Roberto
Chrysomya ( Diptera: Calliphoridae) in the New
B. Monzon, Mahidol
World. J. Med. Entomol. 21: 105- 113.
University ( Thailand);
Demei
Otobed, entomologist for the Republic ofPalau; Vincent

Lee
and
Paul H. Arnaud ( CAS), Norman
NMNH), Steve Kutcher (

megacephala
who
in California),
Woodley
first discovered C.
and
the many
other
colleagues and coworkers whose encouragement and
Baumgartner, D. L. and B. Greenberg. 1985. Distribution

and medical ecology of the blow flies ( Diptera:
Calliphoridae) of Peru. Ann. Entomol. Soc. Am.
78: 565- 587.
support helped with the development of this review.
Bennett, S. G. 1989. New fly species invades southern

California. Vector Ecol. Newsl. 20(4): 15.
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DECEMBER, 1993
PRELIMINARY RESEARCH ON BACILLUS THURINGIENSIS VAR. ISRAELENSIS

USE FOR MOSQUITO CONTROL IN NORTHEAST GREECE
K. Ouzounisl and A. Samanidou- Voyadjoglou2
ABSTRACT: Northeastern Greece contains four of the eleven large wetlands of the country that are protected by
the Ramsar Convention. Malaria was once a serious health problem, but intensive antimalarial measures eradicated
the disease in 1965. DDT residues are still traceable in many components of the food chain. The destruction caused
in the sensitive ecosystems by the chemicals and plans for tourist promotion of landscapes prompted us to adopt
control measures that would be less harmful to the environment. A pilot study using biological control with Bacillus
thuringiensis var. israelensis( Bti) began in the Summer of 1992. Results revealed 95- 100 percent mortality in the
2nd and 3rd larval instars 24 hours following the application of Bti in concentrations of 250 gr/ha. In a parallel study
on mosquito species, 27 species in five genera were found in the area.
INTRODUCTION
Bacillus thuringiensis var. israelensis( B ti) for mosquito

control was explored. Bti was selected because of its
The
coastal
by
characterized
about
region
of
Northeast
Greece,
an abundance of wetlands, stretches
200 km along the sea and 10 km inland. Two large

the Nestos and Evros, and numerous streams
( Thomas and Ellar 1983, Margalit and Dean 1985). The
Several lakes, marshlands, and lagoons
against diseases transmitted by mosquito vectors while

protecting the diverse biological organisms inhabiting
rivers,
traverse the area.

also exist,
45
Km2),
the most important of which are Lake Vistonis
lagoons in the Nestos
and
150 Km2). Another factor contributing

extensive
1960s for the
irrigation
agricultural
system
the wetlands and promoting tourism in the area. Also,
Evros Delta
it was deemed necessary to develop a new list of

mosquito species inhabiting the area because the existing
information( e.g. Pantazis 1935, Livadas and Spangos
entire area has a considerable potential
development, but this is
to the wetlands
in the
constructed
development
the area. The
1940) was considered outdated after the tremendous
for tourist related
environmental alterations of the area since the second
by
prohibited
primary aim of this research was to protect the public
15 Km2),
the complex of the Lakes Metrikou(
and numerous
is the
selective activity against mosquito larvae and its

successful use in many countries for more than a decade
of
the tremendous
World War.
mosquito populations that emerge from numerous

sites created
in the
was once the most serious public
health
temporary breeding
permanent or
wetlands.
Malaria
problem
in this
part of Greece with thousands of
human
especially DDT
Tests of Bti were conducted in five natural breeding

places: a coastal marsh( I), a draining stream( II), a lake
in the woods with stagnant water( III), temporary lakes
overflowing after heavy rain( IV), and a rice field( V).
for
The size of the treated habitats and their water
annually ( Livadas and Spangos 1940).

Chemical insecticides, including chlorinated hydrocases reported
carbons and
were
the
malaria.
Since 1972, the
prohibited,
in mother'
et al.
organophosphates,
first intensive
vector
and
control
use of DDT
but high levels
measures
in Greece has been
Two different preparations of Bti were used, derived
indices( Ouzounis
from a dry powder formulation ( potency 2,500 ITU/

mg), depending on the vegetation type of the treated
1985, Ouzounis 1990). Also, mosquitoes developed
resistance to most of the chemical
characteristics are summarized in TABLE 1.
detected
of residues are still
s milk and other biological
Test of Bti
insecticides
used
habitat. These included a water solution( 250 g Bti/ 101
using
rate of 0.9 Kg Bti/25 Kg dried sand/0.7 1 seed oil. The
WHO 1986).
H2O) and a mixture with fine granule sand( 1 mm) at a
In this preliminary
work,
the
possibility
of
1Democritus University of Thrace, 681 00 Alexandroupolis, GREECE.

2Athens School
of
Public Health, Athens, GREECE.
148
TABLE 1.
DECEMBER, 1993
Physical characteristics of the five habitats at the time of Bti treatment.
Habitats Created
Physical Characteristics
m2
surface size/
depth/ cm
II
III
20000
200
100
500
5000
40- 60
30
50
10- 15
10- 20
larvae/ 1 H2O
pH
air temperature
D. O. mg/ 1
D. O. percent
mS/ cm
conductivity
salinity/
150
20
50
10- 15
10- 20
8. 30
8. 32
7. 62
8. 81
8. 49
30
30
18. 5
19. 1
32
27. 5
25
17. 4
23. 8
30.2
10. 5
4. 9
1. 8
10. 6
9. 1
133
62
29
130
120
5. 65
5. 09
0. 75
8. 61
0.32
3. 5
3. 1
0. 5
5. 4
0. 1
temperature
water
IV
00
Habitat I: coastal marsh
II: draining stream

III: lake in the woods
IV: temporary lakes

V: rice field
first
preparation
habitat IV),
in the
second one
abundant.
was applied
The
July- August
habitats
other
was a
1991
and
located in villages covering a total area of 10,000 ha.
where vegetation was
Mosquitoes were collected with an aspirator from the
the time of the
darkest sites of the dwellings, e.g. corners of the ceilings,

where they were resting during the day. Collections
were made at monthly intervals during 1991- 1992.
Some nighttime collections were also performed during
the summer in 1991 and 1992 by using a human bait
described by Becker and Ludowig( 1989).
Taxonomic determinations were made by using
total of six applications
during
June 1992. The proximity
May-
habitats treated was approximately 1- 4 Km

from dwellings and other inhabited areas.
of mosquito
The larvae
their natural
used
in the tests
breeding
adult mosquitoes. These were stables or sheepfolds
vegetation, and the
material was prepared at
There
application.
in the temporary lakes
lacking
which were
were collected
places prior
from
to Bti treatment,
transferred into enamel pans, and separated
by
instar
keys of Du Bose and Curtin( 1965) and Betzios( 1989).
and genus. They were then transferred to floating sentinel

breeding places. Twenty- five larvae of 2nd
and 3rd or early 4th instar were introduced into each
sentinel cage immediately after Bti treatment. Three to
cages at the
five cages,
spaced
larval habitat,
30 m apart, were introduced into each
depending
Containers of equal
filled
with
in TABLE
an equal number of
Larval
1.
density
and
V,
and
of
Bti
these habitats is
Anopheline larvae
used
of
the
in the tests conducted
Aedes larvae
tests conducted in the other
of
prior to
4th instar larvae. No differences were observed in the
same
Maculipennis complex were
Study
Both formulations of Bti were highly effective against
habitat
in a protected place near the treated habitats
containing
to serve as control.
in habitats I
introduction of the sentinel cages in the treated habitats.
the sentinel cages half
2nd and 3rd instar larvae with mortality of 95- 100

percent. Mortality ranged from 80- 90 percent in early
treatment and
given
the treated surface area.
volume, with
from the
were used
in the
habitats.
sites were selected
effectiveness ofB ti in the five treatedhabitats. Relatively

high mortality of about 11. 2 percent was achieved in all
the control tests, which was probably due to
environmental changes during transportation and
maintenance of larvae in the control containers.
Mosquito Species
Sixteen
Bti Efficacy
Larval mortality was recorded 24 hours after
larvae,
water
were placed
on
Mosquito Species
for the collection
of
The results regarding the mosquito species occurring
DECEMBER, 1993
in the
July
area are presented
and
August, the
collections was
in TABLES 2, 3,
and
most prevalent species
Anopheles
maculipennis
4.
incidence of all mosquito species, and especially ofAn.

maculipennis, was reduced. This was probably due to
the fact that irrigation canals and draining streams had
dried up after crops had been harvested. Several marshes
During
in daytime
followed
by
Anopheles sacharovi. The latter species is found mainly

in
brackish swamp areas. Culicine mosquitoes

rarely found in dwellings. In September the
and swamps in the area dried because of high summer
coastal
were
TABLE 2:
149
temperatures and drought. By the end ofMarch, mosquito
Cumulative list of anopheline mosquito species. Daytime
collections from dwellings at monthly intervals during 19911992.
Species
Relative abundance*
Anopheles
maculipennis
Anopheles
sacharovi
Anopheles
superpictus
Meigen
abundant
Favre
abundant
Grassi
moderate
Anopheles plumbeus Stephens
rare
Anopheles
claviger
rare
Anopheles
algeriensis
abundant:>
Meigen
Theobald
rare
50 mosquitoes in average in 15 min.
moderate: 25- 50 mosquitoes in average in 15 min.

rare:<
25 mosquitoes in average in 15 min.
TABLE 3.
Cumulative list of Culicine mosquito species. Daytime
collections from dwellings at monthly intervals during

1991- 1992.
Mosquito species
Aedes caspius*
Aedes cantans( Meigen)
Aedes vexans( Meigen)
Aedes geniculatus( Olivier)
Aedes annulipes( Meigen)
Aedes pulchritarsis( Rondani)
Culex pipiens( Linnaeus)

Culex modestus( Ficalbi)
Culex laticinctus( Edwards)
Culex univittatus( Theobald)
Culex hortensis( Ficalbi)
Culex impudicus( Ficalbi)
Culex mimeticus( Noe)
Culex territans( Walker)
Culiseta longiareolata( Macquart)
Culiseta fumipennis( Stephens)
Culiseta annulata( Schrank)

Coquillettidia richiardi( Ficabli)
rare;<
25
mosquitoes
in
average
in 15
min.
150
TABLE 4.
DECEMBER, 1993
Cumulative list of nighttime collected mosquito species with
human bait during summer 1991 and 1992.

Mosquito
Relative abundance
species
Aedes
caspius(
Pallas)
Aedes
vexans(
Meigen)
Aedes
cantans(
Aedes
geniculatus(
Aedes
echinus(
abundant*
moderate**
Meigen)
rare***
Olivier)
moderate**
Edwards)
rare***
Culex pipiens( Linnaeus)

Culex
Anopheles
abundant*
Ficabli)
modestus(
abundant*
Meigen)
maculipennis(
rare***
50 mosquitoes in 15 min.
25- 50 mosquitoes in 15 min.

25 mosquitoes in 15 min.
densities began to increase. Anopheline species became

abundant
in the
July( TABLES
middle of
Human baits
were
used
for
and
outdoor
3).
evening
5.
Aedes vexans was a common species, frequently

collected from human bait especially in inland
areas.
collections performed in different inhabited places

between 8: 00 PM and 11: 00 PM at 30 min. intervals.
Acknowledgments
Results are shown in TABLE 4. Aedes caspius was the

predominant
human
biting
Culex pipiens, Culex

also
very
species
modestus and
frequently collected
in the
coastal areas.
Aedes
in the
This project was subsidized by a grant from the
vexans were
District of East Macedonia and Thrace. The cooperation
whole area of our
of Dr. N. Becker with the Bti application is greatly
study.
appreciated.
CONCLUSIONS
REFERENCES CITED
1.
The experimental application of B ti was limited

to small scale field trials because it is not yet
registered
for
use
in Greece.
Becker, N. and H. W. Ludwig. 1989. Microbiologische
Stechmuckenbekampfung. BiologieinUnsererZeit
4: 10- 111.
2.
Anopheles maculipennis was the predominant

anopheline species
densities
maximum
in the area
and attained
during July
and
August.
Betzios, V.
1989.
Arthropods of public Health.
Agrotechniki", Athens, 260 pp. (In Greek).
This period coincides with intensive irrigation

in the
3.
Du Bose, W. P. and T. J. Curtin. 1965. Identification

keys to the adult and larval mosquitoes of the
area.
Anopheles
sacharovi
followed
An.
Mediterranean area. J. Med. Ent. 1: 349- 355.
maculipennis in frequency during July and

August. Both
species were rare
in
winter.
Livadas, G. and J. Sphangos. 1940. Malaria in Greece.

Vol. I. "
4.
Pyros" press Ltd, Athens, 248 pp.
Aedes caspius was the predominant culicine

species
in
frequently
coastal areas and the species most

collected
from human baits. Culex
Livadas, G. and J. Sphangos. 1940. Malaria in Greece.
Vol. II. "Pyros" press Ltd, Athens, 300 pp.
pipiens also prevailed in large numbers in the

area.
Margalit, J.
and
D. Dean. 1985. The story
of
Bacillus
DECEMBER, 1993
thuringiensis var.
israelensis. J. Am.
Mosq. Cont.
151
1578.
Assoc. 1: 1- 7.
Pantazis, G. 1935. La faune des Culicides de Greece.
Ouzounis, K.
1990.
Biocides in
mother' s
milk.
VI Congr. Int. Ent. Madrid, pp. 911- 936.
Conference on chemicals in the environment. Univ.

Aege. Molivos, Lesbos, Greece. Sept. 3- 6. pp.
503- 508.
of
Thomas, W. E. and D. J. Ellar. 1983. Mechanism of

action of Bacillus thuringiensis var. israelensis
insecticidal- endotoxin. FEBS Letters 154: 362-
Ouzounis K., K. Oxynos, I. Gebefugi,
and
M. Bahadir.
368.
1985. Vorkommen chlorierter Kohlenwasserstoffe

in
ausgewaehlten
chenlands (
Umweltproben Nordgire-
Xanthi). Chemosphere 14 ( 10): 1571-
WHO. 1986. Resistance of vectors and reservoirs of

disease to pesticide T.R. S. No 737. Geneva, 87 pp.
BULL. SOC. VECTOR ECOL, 18( 2): 152- 159
DECEMBER, 1993
MOSQUITO UTILIZATION OF RESTING SITES AT

AN URBAN RESIDENCE IN SOUTHERN CALIFORNIA
E. T. Schreiber1, 2, W. E. Waltonl t' 3, and M. S. Mullal
ABSTRACT: We examined the distribution of adult resting mosquitoes at 30 sites surrounding a residence in the
urban Los Angeles area. The distribution of adult resting mosquitoes remained remarkably constant throughout the
five month( June- October) study; resting adult mosquitoes consistently preferred sites on vertical surfaces above
decaying vegetation. The number of mosquitoes at resting sites was greatest at midday( 1100- 1300 PST) and was
less at sampling periods in the morning( 0700-0900 PST) and late afternoon( 1400- 1600 PST). The abundance of
resting adult mosquitoes at this residence was greatest in June and was lowest in August. Resting mosquitoes were
predominantly males( 61% of the population) and the preponderance of males in AFS Sweeper samples may reflect
the proximity of a larval developmental site to the residence. Biotic factors, such as predators, non- culicine dipterans
chironomids), or other mosquito species did not explain the observed distribution of the dominant mosquito, Culex
quinquefasciatus, and the distributions of three less abundant species. Differences among sampling sites for abiotic
factors, such as light intensity, ambient air temperature, and relative humidity did not satisfactorily explain the
observed distributions of resting mosquitoes; however, the importance of within- site differences ( microscale
differences) in these factors may have been underestimated.
INTRODUCTION
of bright sunlight and/ or on relatively open, low

vegetation, Schreiber et al.( 1989) postulated that light
Studies
mosquitoes
of
in
the distribution
nature are
intensity was an important determinant of mosquito
of
resting adult
limited. Service( 1969, 1971)
believed that the location
of
hosts
distribution, especially for Culex quinquefasciatus Say,

in urban, residential yards. Light intensity was also
was not responsible
for the patchy distribution of bloodfed mosquitoes in

Britain, but that the selection of resting sites was strongly
influenced
by
Previous
shade.
Bidlingmayer( 1969)
and
studies
by
Service ( 1969)
Edman
and
noted to be of significance for Cx. tarsalis, Culex

stigmatosoma Dyar, An. freeborni, Culiseta inornata
Williston), and Culiseta incidens ( Thomas) in the
selection of red boxes in Fresno, California( Gjullin et
showed that
bloodfed
adults may fly considerable distances before

daytime resting sites were located.
Abiotic factors are thought to have a greater
al. 1963).
The distribution of resting mosquitoes around urban

residences is highly clumped. Reisen et at.( 1988) found
influence on the distribution

than do
that the distributions of several mosquitoes around
of resting adult mosquitoes

biotic factors. Although Harwood and Halthill
1960) found that
arachnid predators
urban residences within Los Angeles County fit a
deterred Culex
negative
binomial
model.
An analysis of the spatial
tarsalisCoq. fromresting, Schreiberet al.( 1989) reported
distribution of resting adult mosquitoes at two urban
that predators did not
residences showed that mosquitoes were aggregated
mosquitoes.
intensity
As
significantly influence resting

biotic interactions, light
compared to
was of greater
importance in the
selection of
resting sites by Cx. tarsalis and Anopheles freeborni

Aitken adults in California( Harwood and Halthill 1969).
Because mosquitoes were never found resting in areas
and the areas ofconcentration were temporally consistent

(
Schreiber
et al.
1989).
Furthermore, Schreiber et al.
1989) found that populations of resting mosquitoes

around urban residences were predominantly males.
Here we report on a five-month study, which was
tDepartment of Entomology, University of California, Riverside, CA 92521, U.S. A.
2Current address: Florida A&M University, John A. Mulrennan Sr. Research Laboratory, 4000 Frankford Ave.,
Panama City, FL 32405, U.S. A.
3Current address: College ofLife Sciences and the Department ofZoology, University ofMaryland, College Park,
MD 20742, U.S. A.
factors regulating the
distribution
spatial
and species
F tests were calculated. Goodness of fit tests for linear
to examine
and quadratic trends were also run for the time of day
mosquitoes
effect after inspection of the variance- covariance
composition of resting adult mosquitoes and
the differences in the distribution of resting
during daylight hours.

influence resting
B y understanding the factors that
mosquito abundance and
this work may then enable better
landscape
via
statistical significance of comparisons and approximate
to elucidate the
residence,
urban
at one
undertaken
To determine whether biotic or abiotic factors
distribution,
significantly influenced resting site preferences of
management strategies
by
and
architecture
matrices.
mosquitoes, we examined the relationship of mosquito
adult
reducing
abundance to the other biota captured in AFS Sweeper

samples and to several abiotic factors. For each mosquito
harborage.
mosquito
153
DECEMBER, 1993
species in the collections, five biotic variables were
examined statistically: the presence- absence of

conspecifics, other mosquito species, chironomids,
Adult resting mosquitoes were collected semimonthly from June to October, 1989, at 30 sites
in Irvine, California.
a residence
surrounding
The
predators ( spiders
detailed description
of the
air temperature, and relative humidity were measured
included trees,
i. e.,
sites
Due to a preponderance of zeros in samples, the
structures
data for the biotic factors were transformed to 4X+1
The 30 sampling
divided into five discrete habitats: ( 1)
above
and
under raised patios,
walls,
and under the eaves of the house).

were
flowers,
shrubs, grass, mulch,
man- made structures (
2)
finding if the sexes differentiated among resting sites.

Samples from each site were categorized by the sex of
the mosquitoes captured: ( 1) males only; ( 2) females
only; and( 3) both sexes.
Each resting site was sampled for 90 seconds with

AFS Sweeper( Meyer et al. 1983). Potential resting
sites
decaying vegetation( i.e., compost, mulched beds);
man- made structures without vegetation( i. e., patios,
3) flowers(
eaves, walls);(
backyard
vegetation (
above unmulched
beds);( 4)
trees and shrubs); and (
yard vegetation ( trees and shrubs) (
Fig.
from
were taken
ground level to
m and
the ground surface at three sites
Fig.
vegetation(
Twice
5) front
Vertical
1).
stratification of mosquitoes was also studied.
m above
and other flying
prior to sampling each site. We were also interested in
vegetation.
an
ants),
Three abiotic factors were
each of the 30 sampling sites. Light intensity, ambient
its
site and
study
and
insects.
measured with a hand- held meter at a central position in
richly landscaped and the verdure was

fastidiously maintained. Schreiber et al.( 1989) provide
residence was
a more
nondipteran)
Samples
from 4
m to
in the backyard
Steele and Tonic 1980) and then analyzed by ANOVA

SAS Institute 1982).
Least square
GLM
LS) means were calculated for each variable and
procedure,
Duncan' s Multiple Range Test was used to compare LS

means. Vertical stratification of resting adultmosquitoes
was analyzed by t- test( Sokal and Rohlf 1981). In order
to examine the relationship between mosquito abundance

and the abiotic factors, mosquito abundance was
regressed on each abiotic factor.
1).
month, all of the
during each
the residence were sampled at
resting sites at
three time intervals:
Changes in faunal similarity for each habitat were
examined using the index of interassociation ( Pielou
Twenty-
1974). The chi- square statistic was used to test if species
four hours separated the collections for each time interval.
assorted nonrandomly among resting sites. If species

assort independently, then the index of interassociation
0700- 0900, 1100- 1300,
and
1400- 1600 PST.
The order at which the three intervals were

at random
chosen
for
each
period,
sampling
with
samplings
sequential
eliminating
sampled was
thus,
temporal
will not differ significantly from the critical value of the

chi- square test.
bias.
A
repeated measure
ANOVA
was used to test
RESULTS
for
differences in the average number of mosquitoes

collected
Because
among resting
deleted from the

mosquitoes
to
in
beds
on most
analysis.
analysis.
day)
Culex quinquefasciatus was the most abundant
in the flowers
mosquito species and comprised 63 percent of the total
The
Habitat
and the
within- subject
was
catch(
TABLE 1).
number of
soma(
13%),
dates, this habitat

average
a habitat was transformed to
between- subject factor. Two

time of
habitats).
categories (
mosquitoes were not collected
above unmulched
prior
site
was
log+
examined
value
as
main effects( month and
interaction terms
factors. Pillai' s trace
were
tested as
was used to test the
and
Culex
tarsalis(
18%), Cx. stigmato-
Cs. incidens( 6%) were relatively rare.
The relative abundance of the mosquito species changed
very little among the time periods ( averaged across

Culex quinquefasciatus comprised 41 percent
of the individuals collected in June and was Z 66 percent
months).
of the mosquitoes collected thereafter(
TABLE 2).
154
BULL SOC. VECTOR ECOL.
DECEMBER, 1993
D)
14;.; .
0
0
'
FRONT
CC ;
ofSe. . ....
8>
1:3
C
o
0 CS
CD :
z as
rij a
crs 45
E o
tri co . >
a)
E ....
It15i:'
N-1
0)
u)
o
Mitgion
I
N
N
0
0
0
0
1>
as
ON:
3
gtirg!'
IA.
Lk.
v.......-
v...;
laiiii0'"
v.
i
c...
is..
c
c.;:
7*
0
0.
1
Figure 1.
40L" .Cn
i: k.
BAC K
Location ofstratified
sample samples
sites surrounding a residence in Irvine, California( A-Z and FF, QQ, ZZ, WW).
Vertically
were taken
from 0 to 1
m and
4 to 6
m at sites
D, E,
and
F.
155
DECEMBER, 1993
TABLE 1.
Percentages of resting mosquito species and males collected at three discrete time intervals
at the Irvine, California site in 1989.
Species
Culex
Culex
Culex
Culiseta
quinquefasciatus
tarsalis
stigmatosoma
incidens
Time
N1
period
Males
0700-0900
131
63. 4
57. 3
18. 3
16. 0
8. 4
1100- 1300
168
61. 3
64. 9
17. 9
12. 5
4.8
1400- 1600
104
58. 7
65. 4
19. 2
8. 7
6. 7
Total
403
61. 3
62. 5
18. 4
12. 7
6.4
Total number collected.
Percentages of resting mosquito species and males collected monthly at the Irvine, California
TABLE 2.
site in 1989.
Species
Culex
Culex
Culex
Culiseta
Males
quinquefasciatus
tarsalis
stigmatosoma
incidens
June
64. 8
40. 7
27. 1
18. 6
13. 6
July
84. 2
65. 6
14.4
8. 9
11. 1
August
61. 7
74. 2
17. 7
8. 1
0. 0
September
58. 1
65. 1
14. 3
19.0
October
51. 9
78. 6
17. 9
3. 6
Time
period
The
percentage of males
was, on average,
of males
in
in AFS Sweeper
samples
61percent( TABLE 1). The proportion
samples
did
not change
However,
appreciably across
between 62
1. 6
0
diel collections( F1. 12= 4.65, P= 0.046). The abundance

ofresting adult mosquitoes in midday( 1100- 1300 PST)
collections was, on average, greater than in either
in
morning or late afternoon sampling periods for every

month except September( Fig. 2).
in
In general, abiotic factors ( measured at a central
collections declined through the summer but was always
position at each sampling site) did not explain the

observed distribution of resting mosquitoes. Except for
the time periods.
84
percent and
percent of the
June through August.
greater than that
The
males were
individuals
proportion
collected
of males
for females( TABLE 2).
Resting adult mosquitoes were vertically stratified.

Significantly more individuals were present in the lower
stratum(
4-6
m:
0- 1
m) than were present
3. 678, P < 0.002).
is =
Males
P < 0.07); however,

in the
were collected
and
mosquitoes at
ts=
resting
among
0. 024).
in
the slope
2. 541,
regressions for mosquito abundance versus abiotic
factors did not differ significantly from zero.

The species composition at resting sites within each
patterns of mosquito abundance
of the five habitat categories was unpredictable( TABLE
sites
ts:
P> 0.05).
number of
declined from June
aspirator samples
months(
P < 0. 01),
estimates for 95 percent ( 37 of 39 regressions) of the
until
August and increased thereafter( Fig. 2). The number of

mosquitoes
related to mosquito abundance (
sex
upper stratum(
diel
both temperature and relative humidity were significantly
more
equivalent numbers of each
the Irvine site. The average
were evident at
stratum
were
females in the lower stratum(
abundant than
Seasonal
in the higher
late afternoon ( 1400- 1600) samples in August when
differed significantly
F4, 14= 3. 93, P=
Pillars trace: Approx.
significant quadratic
trend was evident for
3).
In more than half of the time periods, the index of
interassociation did not differ significantly from that

expected if the species composition was determined by
chance alone. Whereas, the indices of interassociation
suggest that species were not assorting independently

among resting
sites
during the early afternoon( except in
156
DECEMBER, 1993
June
July
5
Aug
8
Sept
a\
Oct
_
1
07: 00- 09: 00
11: 00- 13: 00
14: 00- 16: 00
Time of Day
Figure 2.
The average number of mosquitoes collected in AFS Sweeper samples from all habitats from June until
October 1989.
August), the majority
of
morning
and
late
E, and F) and on vegetation ( ivy) near the mulch pile
afternoon
comparisons suggest that any particular mosquito species
site A).
was not differentiating among resting sites. However,

in
order to meet the assumptions of the chi- square test,
the numbers of individuals of the three rare species in

samples
had to be
pooled
Particular resting
in
most
instances.
sites were preferred
by resting
adult mosquitoes and maintained their attractiveness to
DISCUSSION
The nonrandom vertical and horizontal distributions
of adult mosquitoes surrounding an Irvine residence

suggested that mosquitoes distinguish among potential
resting sites. Resting adult mosquitoes clearly preferred
the resting mosquito population. While the number of
vertical structures that were near the soil, were shaded,
individuals
and might have been comparatively humid. For example,
collected
significantly
during the
mosquitoes at
was
in AFS Sweeper samples changed
resting
five
sites
months,
the distribution of
the greatest concentration of resting mosquitoes was
the residence
found on the vertical walls S 1 m above decaying
surrounding
surprisingly constant throughout our study(
The greatest numbers
Fig. 3).
vegetation( site B). The abundance ofresting mosquitoes
of resting mosquitoes occurred on
in AFS Sweeper samples from this site remained high
shaded, vertical surfaces: (

pile( site
1)
a wall above a
B);( 2) in flower pots that
decaying leaf litter( site G);( 3)

a mulched
bed(
site
FF);
leaf mulch
contained a
layer
of
the base of a trellis above
4)
until the compost pile was removed in October by the

homeowner.
Resting adult mosquitoes were also
concentrated on flower pots that contained decaying
under the eaves of the
vegetation, on a shaded, vertical surface above a mulched
front entryway( site S). Resting mosquitoes were also

collected from three large trees in the backyard( sites D,
flower bed and, to a lesser extent, on a sheltered vertical
and(
surface
beneath the eaves of the residence. The backyard
DECEMBER, 1993
157
TABLE 3. The Index of Interassociation for resting mosquitoes calculated by month

and time period.
Month
Time
June
0700- 0900
10. 52**
1100- 1300
81. 03***
1400- 1600
6. 02*
July
August
September
October
d. f.t
period
2$
0700- 0900
1100- 1300
1400- 1600
0700- 0900
1. 32 NS
1100- 1300
3. 69 NS
1400- 1600
1. 40 NS
0700- 0900
5. 99 NS
1100- 1300
6. 86*
1400- 1600
0. 33 NS
0700-0900
1100- 1300
1400- 1600
3. 93 NS
10. 94**
3. 75 NS
1. 07 NS
13. 17**
0. 48 NS
td.f.were first pooled by number of species so that not more than one of the expected
frequencies was less than five.
Goodness
of
fit test
where
NS
P 50. 01; *** =
0. 005 <
means not significant at
0. 001 <
P 50.005.
P> 0.05;*=
0. 01< P 50.05,
H0: species are independently
distributed.
was more shaded than
the front yard and the
vegetation
in the backyard contained more resting mosquitoes
metropolitan area(
Schreiber
et al.
1989). Although the
than
distribution of all mosquito species among the resting
apparently prefer particular sites

and move among resting sites during daylight hours;
movement among resting sites is not restricted to
attractiveness to mosquitoes. The species composition
did the
vegetation
Adult
in front
of the
house.
sites suggests that resting sites differ predictably in their
mosquitoes
crepuscular
periods.
We
sampled
at any particular resting site can change dramatically

from
one sample to the
next.
The apparent lack of
adult
partitioning of sites among the species suggests that
mosquitoes without replacement yet subsequent samples
resting sites are not limiting, and these species are not
competing for resting sites around this urban residence.
at time periods within each month and
unambiguously
resting
among
months
show that mosquitoes congregated at
the same resting sites(
Fig. 3). The
The resting sites examined in our study may have been
in June and
underutilized or may not have had the highest

concentration of resting adults at sites within the Irvine
between 1100- 1300 PST during four of the five months.
neighborhood. Additional larger-scale, intensive studies
The diel differences in AFS Sweeper
are required to resolve such questions.
mosquitoes were collected
greatest numbers of
from resting
sites
catches suggest
that adult mosquitoes were moving among resting
sites
Males and females also did not differentiate among
during the day.

Resting sites do not appear to be strongly partitioned
resting sites. Males were more abundant in lower stata

than were females, but the majority of individuals
among the five mosquito species. Culexquinquefasciatus

was the dominant species collected at the Irvine residence
collected at the
this study) and at other residences in the Los Angeles
Irvine
residence were males.
The
preponderance of males in the resting population may

indicate
developmental
site was
in
close
proximity to
DECEMBER, 1993
158
1-
O
o
a)
1. 4
v o
o)
13
to
to
a)
>,
CD
0) ao
a)
a)
f0
E C
Z.
E
as
y l..
CD
ti
Qtola
CIEBEND
I
l
0
Potemo
I
0
Lt)
0
V
u7
LO
cn
a)
a)
LL.
O)
gg
1
D)
O_
el
t -
Mr
f-
z
J
CO
C7
r._Y
r m
yiff::::rS:i:; S:::=:::
s: b:>:.:
Fir:2. 2:: ir :
O -
- tA
0I
U)
r
O
V
It)
Co -
ig5n1?j.
ioqumN a5B.)aAV
Figure 3.
Abundance of adult resting mosquitoes collected from June until October 1989 at sampling sites
surrounding a residence in Irvine, California. The numbers of individuals collected at each site were
summed across time periods
in
each week and then averaged
for
samples taken within each month.
Schreiber
the residence.
percent of
159
DECEMBER, 1993
et al. (
the individuals
1989) found
85
that
collected around residences
breeding/ developmental
freeway drains were males.
near mosquito
sites, such as
Although abiotic and biotic factors failed to explain

the clumped distribution
resting
adult mosquitoes, a
single measurement for each abiotic
factor at a particular
of
sampling site may have been insufficient to resolve any
Gjullin, C. M., T. D. Mulhern, and R. C. Husbands.
1963. The daily resting cycle of several species of
mosquitoes. Mosq. News 23: 203- 210.

Harwood, R. F. and J. E. Halfhill. 1960. Mammalian
burrows and vegetation as summer resting sites of

the mosquitoes Culex tarsalis and Anopheles
freeborni.
Mosq. News 20: 174- 178.
microscale preferences exhibited by the mosquitoes.

Measurements
at a central
location
maybe sufficient to
discern large- scale differences in abiotic factors among
sampling
sites,
but future
efforts should concentrate on
the measurement of microscale differences in
factors
within, and
sites.
among, resting
abiotic
Gardening by
Meyer, R. P., W. K. Reisen, B. R. Hill, and V. M.
1983. The " AFS Sweeper" a battery
Martinez.
powered backpack mechanical aspirator for

collecting
adult mosquitoes.
Mosq. News 73:
246- 250.
the homeowner ( i.e., removal of the compost in the

B) unmistakably influenced the
Pielou, E. C. 1974. Population and Community Ecology:
distribution of adult resting mosquitoes. The distribution
Principles and Methods. Gordon and Breach Publ.
of sites
vicinity
and
diel
and
movements of adult mosquitoes around the
Inc. New York, NY, 424 pp.
Irvine residence suggest that surface temperatures and

edaphic
factors may play
pivotal roles
in resting
site
To
understand
the behavior of adult mosquitoes in
have
urban settings, we
mosquito
Reisen, W. K.,R. P. Meyer, V. M. Martinez, O. Gonzales,

J. J. Spoeheland,
selection.
distribution,
several residences (
detailed
emphasized
abundance, and
Schreiber
studies of
biting
habits
at
1989, this study).
et al.
and
J. E.
Hazelrigg.
1988.
Mosquito abundance in suburban communities
in Orange and Los Angeles counties, California.
Proc. Calif. Mosq. Vector Contr. Assoc. 56:

75- 85.
The distribution of resting mosquitoes around urban

is apparently predictable and, therefore, has
component yet the factor( s) used
deterministic
strong
residences
SAS Institute. 1982. SAS Procedures Guide. 5th ed.
SAS Institute, Cary, NC, 584 pp.
by individuals to select resting sites remains to be

The
determined.
population
at
resting
Schreiber, E. T., J. P. Webb, Jr., J. E. Hazelrigg, and M.
remarkably
at individual
associated with urban residential areas in Los
Angeles Basin, California. Bull. Soc. Vector Ecol.
species composition of the
the
Irvine
residence
was
constant even though species composition

sites around the residence was much
better understanding
of
less
predictable.
the mosquito ecology/ behavior
S. Mulla.
1989. Bionomics of adult mosquitoes
14: 301- 318.
in these functional units( sensu Urban et al. 1987) may

permit the more effective
modeling
disease transmission
and
populations
and management of
pestiferous
in the neighborhoods
and,
mosquito
eventually, in the
Service, M. W. 1969. Observations on the ecology of

some Britain mosquitoes. Bull. Entomol. Res. 59:
111- 194.
entire metropolitan area.
Service, M. W.
1971.
The daytime distribution of
mosquitoes resting in vegetation. J. Med. Entomol.
Acknowledgments
8: 271- 278.
This research was supported by the University of

California Special Funds for Mosquito Research and the
Orange County Vector Control District. Thanks to G. L.
Sokal, R. R. and F. J. Rohlf. 1981. Biometry. W. H.

Freeman and Co. San Francisco, CA, 859 pp.
Challet for the use of his residence and Dr. J. A. Gruwell

for
assistance
Steele, A. G. D. and J. H. Tome. 1980. Principles and
in the field.
Procedures
REFERENCES CITED
Edman, S. D.
and
of
W. L. Bidlingmayer.
blood- engorged
capacity
News 29: 386-392.
of
Statistics.
McGraw Hill Book
Company, New York, NY, 632 pp.

1969. Flight
Urban, D. L., R. V. O' Neill, and H. H. Shugart, Sr.
Mosq.
1987. Landscape Ecology Bioscience 37: 119-
mosquitoes.
127.
DECEMBER, 1993
FIELD TRIALS OF A FIZZY TABLET WITH BACILLUS THURINGIENSIS

SUBSP. ISRAELENSIS IN FOREST SPRING PONDS IN DENMARK
O. Skovmandl and A. G. Eriksen'
ABSTRACT: A fizzy tablet consisting of a spray-dried powder ofBacillus thuringiensis subsp. israelensis( Bti) was
constructed to obtain a product for mosquito larval control that was easy to store, carry, and use. The formulation
was tested in the early spring of 1991 in 10 small, temporary spring ponds containing mosquito populations ofAedes
cataphylla and Aedes cantans. Half the tablet dose per 5 m2 killed 85 to 88 percent of 2nd to 3rd-instars. A full tablet
dose per 5 m2 killed 93 to 100 percent of 2nd to 3rd and early 4th-instar larvae in ponds without dense vegetation,
and 75 to 82 percent of the same instars in ponds with dense vegetation. In ponds tested later in the season and
consisting nearly of only 4th-instar larvae, the effect was 73 to 79 percent. Dense vegetation or a large amount of
dead leaves on the surface prevented the particles from spreading. The tablets are thus most usable in breeding sites
without these obstacles, such as water reservoirs, garden pools, swimming pools, and small streams. B ti does not kill
late 4th-instar larvae because they do not ingest it.
INTRODUCTION
Products based on Bacillus thuringiensis subsp.

israelensis ( Bti) have been
mosquito and
black
DeBarjac 1978, Davidson
for the
used
larvae for
fly
Sweeney
and
control
of
Product Formulation
10
years
The fizzy tablets were produced by mixing Bti
1983).
The
primary powder, detergents, inert powder, and citric

acid with calcium hydrogencarbonate to allow the
more than
insecticidal property of these preparations is due mainly

to the proteinaceous crystal, the delta- endotoxin
spreading of the tablet ingredients. Preliminary trials in
during the sporulation phase of the bacterium.
ponds and streams assessed various formulations that
produced
The
production of
B ti is based
process,
a concentration
followed
by
either
formulation
of the
spray- drying
and
and
crystals
formulation
of the concentrated cream.
obtained are similar to traditional
fermentation
on a
spores
The
or a
products
flowable concentrates
resulted in the tablets disintegrating in less than a minute

and dispersion over approximately 2 m2 but that did not
disintegrate during transport. Thepercentage ofprimary

Bti powder was adjusted to construct tablets with a
potency of approximately 750 ITU/mg.
and wettable powders and can be applied with

conventional equipment.
also
formulated by the
be
sand granule" that can

water
applied
by helicopter
over
that is covered with vegetation.
Mosquito larvae
ponds and in
et
The Bti spray- dried powder is
applicator with sand and oil as a
al.
1991).
To treat these
small water
are not
very
convenient.
easy to carry, anddisperse well.
the
sites,
would need no equipment
traditional
A better product
for
application,
showed that the tablets could
described for
a similar
Preliminary
lations in
field trials
be easily
tablet ( Becker et
18 to 160
m2)
fizzy
1991).
al.
were used to compare
in
In this report, we investigated the possibilities of

as a
complete dissolution of the tablet and it was serially

diluted.
( 1984),
In a protocol similar to McLaughlin et al.

20 4th-instar larvae of Aedes aegypti were
added to each cup, with three cups per concentration and

seven concentrations per sample.
IPS82 lyophilized
powder( Institut Pasteur) was used as a standard. LCso

values were computed by probit analysis.
Such a use has also been
small water ponds(
Bti formulated
be
Laboratory experiments
using buckets
used in water
containers.
The toxicity ofa tablet was determined by dissolving

it in 21 of tap water at room temperature with a magnetic
stirrer. A sample was taken from this container after
breed in very
drinking or garden waterreservoirs( Becker
formulations
ideally
often
Bioassay of Toxicity
Ponds
formu-
Eleven temporary water ponds were selected in a
forest.
forest in North Zealand, Denmark, based on their size

( below 200 m2) and the presence of more than two
mosquito larvae per dip with a half liter dipper. The
applying
tablet to meet these criteria.
Novo Nordisk Plant Protection Division, Product Development, 2880 Bagsvaerd, Denmark
between 6 C in
temperature of the ponds varied
12 C in
mornings to
161
DECEMBER, 1993
some afternoons
during
some
disintegrating tablet particles to move unhindered on
April,
the water surface. When this movement was prevented
by grass or dead leaves, the effect was below 90 percent.
1991.
In one pond, one tablet per 5 m2 was used. This resulted

in a control of 83 percent( TABLE 1), but in all other
Sampling
Samples
were taken with a
from the edges,
10 to 20
dipper 30
samples per pond,
according to
Samples were taken before and 24 hours
pond size.
cm and
treatment. The sampled invertebrates were
ponds,
two tablets were used.
In ponds with larvae
earlier than 4th- instar and without too many physical
after
barriers, the control effect was 93 to 100 percent ( 4
to
ponds, TABLE 2). In ponds with more physical barriers,
returned
the ponds after counting, but one to two samples from all
Danish Pest Infestation
the effect
was
75 to 82
percent (
TABLE 2).
In two
Laboratory to confirm species identification. During the
ponds tested at the end of April, many larvae were in the

4th- instar and many pupated during the 24 hour test
counting that occurred in the field, we did not discriminate

between the two dominate mosquito species ( Aedes
percent( TABLE 2).
ponds
were brought to
the
period.
The control of the 4th- ins tar was 73 to 79
cantans andAedes cataphylla), but they were classified

Nontarget Invertebrates
according to larval instar.
Some ponds also contained large populations of
water cricket, Asellus aquaticus, the beetle Haliplus sp.,
Treatment
Ten
ponds were
tablets per
or
m2.
dosed
For their
dead leaves in the
in
use
ponds with
water surface,
dense
dividing
The
and an unidentified oligochaete worm. None of these
grass
groups were affected by the treatments. The larvae of

the chaoborid Monoclonyx sp. was seen in two ponds
before and after treatment but only a few per pond. They
survived for some days after being brought into the
laboratory after treatment, and were then discarded. A
tablets were broken
in half to obtain a better distribution. One pond

as a control.
two
with one( one case) or
was used
area of the ponds were calculated
drawing
scale
by
the pond into simple
of
figures and adding up the total of their areas.

Tablets were distributed manually as evenly as possible.
geometric
few chironomids were also found and in pre- and

post-treated ponds and did not appear to be affected.
The effect of the treatment in a pond was calculated by

DISCUSSION
summing the number of larvae before and after treatment.
The difference divided by the pretreatment number and

multiplied by 100 is shown in the tables.
Fizzy tablets were formulated to obtain a product

that could be used in small water areas such as temporary
ponds in forests or near coasts, pools, or storage
RESULTS
containers for drinking or rain water. The tablets were

not formulated to obtain sustained release, and
Toxicity
The
the tablets confirmed a
accordingly, they could be used in place of flowable
750 ITU/ mg. This demonstrated that the

process did not cause a deterioration in the
concentrate or wettable powder formulations but not
the primary
prolonged effect. The tablets can be distributed manually
laboratory bioassay
of ca
potency
production
activity
of
of
powder.
instead of briquettes and other products made for a
without any special equipment.
The dosage selected for the ponds corresponded to
Mosquito Species
Aedes
ponds
and
found.
in
cataphylla and
only
In the
few
Ae.
cantans
other mosquito
beginning
of
all
the guidelines used for Skeetal FC( 600 ITU/mg, 21/ ha).
were
This dosage resulted in excellent control except when
were
the water surface was subdivided into compartments by

grass or dead leaves leaving some compartments
untreated, or when many late fourth instar were present.
dominated in
species
April, the Aedes spp.
the second to third- instar; Ae. cataphylla were
mostly
in the third and Ae. cantans mostly in the second. At the

end of the month, only 3rd and 4th- instar larvae and
pupae were
Monoclonyx sp., a
found in two ponds.
found. A few larvae
nonbiting chaoborid, were

Chironomid larvae were found in
of
The
must be ingested to have any effect, these larvae will

pupate and survive.
The effect was measured after just 24 hours. The
several ponds.
effect of Bti will most probably increase in the next 24

hours as in laboratory bioassays( data not shown), but
Field Effects
results obtained
dosage, larval instar,
Late fourth instar larvae do not feed, and because Bti
in the
and
ponds
depended
on
the possibilities for the
within 24 hours most other regulating factors may be

ignored. This was confumed by counting in one untreated
162
TABLE 1.
DECEMBER, 1993
Temporary ponds, circular with depth of 10 to 30 cm, area ca 115 m2, were treated with 25 Bti tablets
or ca 1 tablet per 4.5 m2, which is approximately half dosage. The overall result is a reduction of the
larvae population by 83 percent. The raw data are shown to illustrate the inter-sample variation.
Instar
Pretreatment
Sample No.
2.
3.
Post- Treatment
4.
All
3.
0
1
11
10
13
Total
44
59
0. 7
10
Avg./ Sample
6
1
12
3. 7
All
0
0
1
4.
5
6
0.6
1
11
4. 9
Reduction (%)
TABLE 2.
2.
0. 1
0. 5
0. 2
88
85
0. 8
60
83
The table shows the number of larvae caught in three larval stages and the total per pond. The first four
ponds had a water surface without grass or other obstacles and the next three had a water surface full
of grass or dead leaves. These seven lakes were treated early in April when there were very few 4thinstar larvae. The latter two ponds were treated late in April and were dominated by 4th-instar larvae
and pupae( not shown).
Late 4th-instar larvae and pupae do not eat and are therefore not influenced
by the presence of Bti.

Samples
Per
Pond
Pond
Pretreatment
Post- Treatment
2.
3.
4.
All
Reduction
2.
3.
4.
All
(%)
Clear
10
28
168
23
219
13
14
94
Water
10
22
21
47
94
Surface
10
44
25
76
93
Early
20
27
100
Grass
10
44
21
74
and
12
78
77
14
169
15
Leaves
10
26
20
33
79
47
48
10
44
44
13
82
23a
40
76
12
20
75
10
10
79
12
12
73
Early
Clear
Water
Surface
Late
aA sample taken 30 cm from the edge of the pond and the corresponding one taken 1 m from the edge, contained
11
and
6, respectively. The
rest of the samples on the pond contained
to
2.
DECEMBER, 1993
pond
very
close to one of the
test ponds ( data not
163
Bull. Soc. Vector Ecol. 13: 85- 93.
shown). Therefore, the numbers given here may be

Becker, N., S. Djakarta, A. Kaiser, 0. Zulhasril, and H.
regarded as conservative.
Several other invertebrates were also quite common,
including Asellus aquaticus, the beetle Haliplus sp,

an oligochaete worm.
None of these
confirming the already
W.
Ludwig.
1991.
Efficacy of a new tablet
and
formulation of an asporogenous strain of Bacillus
were affected, thus
thuringiensis against larvae ofAedes aegypti. Bull.
well established environmental
Soc. Vector Ecol. 16: 176- 182.
safety of Bti( Ali 1981, Becker 1988). The overall effect

on all ponds treated with two tablets per
control of
95
percent
for 2nd
and
m2
was a
3rd- instar larvae
and
DeBarjac, H. 1978. Dees bacteries contre les moustiques.

Recherches 9: 911- 913.
85 percent for 4th- instar.
Davidson E. W. and A. W. Sweeney. 1983. Microbial

control of Vectors: a decade of progress. J. Med.
REFERENCES CITED
Ali, A. 1981. Bti( ABG 6108)
some nontarget aquatic
Ent. 20: 235- 247.
against chironomids and
invertebrates.
J. Invert.
Pathol. 38: 264- 272.
McLaughlin, R. E., H. T. Dulmage, R. T. Alls, T. L.
Couch, R. I. Rose, and P. L. Versoi. 1984. U. S.
standard bioassay for the potency assessment of

Bacillus thuringiensis serotype H- 14 against
Becker, N. 1988. Mosquito
control
in West
Germany.
mosquito
larvae. Bull. Ent. Soc. Am. 30: 26- 29.
DECEMBER, 1993
OCCURRENCE AND ABUNDANCE OF CYANOBACTERIA IN A BRACKISH

MARSHLAND AND THEIR INGESTIBILITY BY MOSQUITO LARVAE
I. Thiery I, G. Sinegre2, and N. Tandeau de Marsac3

ABSTRACT: A survey of unicellular and filamentous cyanobacteria inhabiting a brackish marshland, located in
the south of France and known to be a mosquito breeding site, was performed over a one- year period and the
abundance of the unicellular strains was compared with the unicellular green algal population that might compete
for the same ecological niche. From each water sample, cyanobacteria were characterized on the basis of their
morphological properties using strains previously purified from the same location as references( Thiery et al. 1991).
The highest densities of unicellular cyanobacteria coincided with the peak season of mosquito larvae which usually
extends from April to October. Each water sample was analyzed under laboratory conditions to determine the degree
of ingestion and digestion of unicellular cyanobacteria by larvae of Culex pipiens and Anopheles gambiae
mosquitoes. Although efficiently ingested and digested by mosquito larvae, cyanobacteria did not appear to be
preferentially selected as a food item compared to other microorganisms present in the natural water samples. This
study showed that unicellular cyanobacteria belonging to the genera Synechococcus and Synechocystis are suitable
candidates for mosquito control, in particular those which, according to the criteria used in this work, are identical
to Synechococcus strain PCC 8905 and Synechocystis strain PCC 8906.
characteristics required to colonize certain habitats.
INTRODUCTION
Consequently, cyanobacteria were isolated anew from

mosquito breeding sites with the aim of introducing
Cyanobacteria are oxygen- evolving photosynthetic

widely distributed in mosquito habitats and
known to be found in the digestive tract of mosquito
prokaryotes
larvae ( Gophen
Khawaled
Bacillus toxins in strains that might be more appropriate
for biological control( Khawaled et al. 1989, Thiery et

al. 1991).
Gophen 1986, Marten 1986,
and
1989). These microorganisms
In 1988 and 1989, unicellular and filamentous
colonize
strains were isolated from a brackish marshland located
the upper layers of bodies of water and could

potentially
on the French Mediterranean coast, which constitutes a
et al.
be engineered to deliver Bacillus toxins in the

areas of mosquito
larvae
feeding
permanent breeding site for Coquilletidia richardii,
over prolonged periods of
Aedes detritus, Aedes caspius, and sometimes Culex
time. Indeed, Bacillus thuringiensis var. israelensis and
pipiens(
Bacillus
to control
currently being
but their limited persistence
unicellular strains were assessed for their ability to be

ingested and digested by larvae of Cx. pipiens and
in the field requires frequent applications( Silapanuntakul
Anopheles gambiae mosquitoes. Due to their easy and
sphaericus are
used
mosquito larva populations,
1983, Davidson
Thiery et al.
1991). After their purification, the
1984, Ohana et al. 1987). In
rapid growth, their tolerance to larvicides and their
an attempt to overcome this problem, toxin genes of
capacity to be efficiently ingested and digested by
et al.
either
B.
were
introduced into
et al.
sphaericus or
B. thuringiensis
var.
israelensis
unicellular cyanobacteria.
recombinant organisms
show
larvicidal
The
activities
Tandeau de Marsac et al. 1987, Angsuthanasombat and

Panyim 1989, Chungjatupornchai 1990).
these recipient cyanobacterial strains
maintained under
ten years and

natural
laboratory
for
or might not possess
larvae,
the
two
unicellular
strains
Synechococcus PCC 8905 and Synechocystis PCC 8906

were selected as the most suitable recipients of bacterial
endotoxin genes.
In this study, the abundance of cyanobacterial

strains morphologically identical to Synechococcus
more than
strain PCC 8905 and Synechocystis strain PCC 8906
in
previously isolated from the same brackish marshland,
the physiological
was compared with that of other cyanobacteria and
conditions
may have lost their ability to
habitats
However,
have been
mosquito
propagate
1Unit6 des Bacteries Entomopathog8nes, Institut Pasteur, 25 rue du Docteur Roux, 75724 Paris Cedex 15, FRANCE.
2Entente Interdepartementale de DBmoustication, BP 6036, 34030 Montpellier Cedex 1, FRANCE.
3Unit6
de Physiologie Microbienne, Institut Pasteur, 28
rue
du Docteur Roux, 75724 Paris Cedex 15, FRANCE.
DECEMBER, 1993
notably from April to October, the
green algae, and
high larval
season of
constitute a nuisance
the degrees
density during
fed
mosquito species
properties.
The cell density of the
filamentous cyanobacteria could not be estimated due to
digestibility of
algae by two
the absence of the formation of discrete colonies or in
on natural water samples were
filaments on the surface of the agar plates. Nevertheless
ingestibility
of
morphological
addition,
cyanobacteria
unicellular
whose cells displayed identical pigmentation and
which mosquitoes
in the south of France. In
165
and
the case of motile strains due to a rapid spreading of the
green
and
the presence and relative amount or the absence of these
compared.
cyanobacteria were noted.
Cells from each colony of green algae were
MATERIALS AND METIIODS
examined under light microscopy, but no attempts were
Sampling
Methods
From
February
made to distinguish the different genera. Abundance

1990
from
samples were collected
Fos/ mer( France).

was collected
immediately
marshland near
Water Sample Analyses
locations, three near the
withdrawn
of
500
analysed
for
Aliquots
mixed.
ml
Water samples were analyzed for physico- chemical
were
parameters that might influence cyanobacterial growth.
physico-
The concentration ( mg/ liter) of nitrate, nitrite,
The remaining 500

flasks, kept in the dark,
ammonium, sulfate, carbonate, and chloride ions was
in less than 24 hours to the Institut
magnesium ions) was evaluated as described ( Rodier
chemical parameters( see
and transported
estimates for the green algae are therefore reported only

for the group as a whole.
water
from one edge. These four samples
and
in
brackish
each of four
were pooled
ml was stored
1991,
February
Every two months, a 250 ml sample
from
edges and one at
until
and
below).
sterile plastic
determined; and the total water hardness( calcium and

1966).
Pasteur, Paris, France.

Water
samples were
ASN- 10
with
hundred l
medium (
medium solidified with
All
Bacto-Agar( Difco).
w/ v)
under
low
photon
flux
density(
plates were
10
Resistivity of the water samples was measured
Rodier 1966).
vegetation),
as
Biotic ( mosquito larvae, fish, and

well
as
abiotic
parameters ( water
were
temperature, pH, and depth) were noted. Precipitation
percent
and day- night variations of temperature were provided

by the nearest meteorological station located in Istres
( France), ca. 10 km from the sampling spot.
of these samples
of each
aliquots
ASN- 10
plated on
3)
serially diluted ( up to 10Thiery et al. 1991). One
incubated
mol/ m2/ s) at
25 C
for three weeks.

Characterization
Cells from
cellular
each
strains)
filamentous
of
or
Feeding Experiment
Twenty-five early 4th- instar larvae of Cx. pipiens
Strains
colony
forming
unit (
filamentous
patches (
strains) were characterized
CFU)(
or
uni-
according to
according to Rippka
and pigmentation,
1979). For this

Optiphot
collected and rinsed in deionized water, crushed in

ASN- 10 medium in a Potter grinder, then centrifuged
et al.
purpose, photomicrographs were taken
contrast
phase
under
illumination
with
photo- microscope equipped with a
55 mm diameter) with 10 ml of each water sample.

After 60 minutes, ten larvae of each species were
morphology( presence or absence of a sheath, gas

vesicles, or baeocytes; cell shape; cell diameter or
cell
width)
strain Montpellier) andAn. gambiae( strain G3), reared

at the Institut Pasteur Paris, were placed in a Petri dish
pseudo-
and treated as
described
by Thiery
et al. (
1991).
One
Nikon
hundred l aliquots of serial dilutions of the treated
Plan 100
larvae and of the water samples, which they were in,
properties of the
were plated onto ASN- 10 agar plates. Incubation, cell
cultured cells were then compared with those of the
counts, observations under light microscopy, and
DL
oil
immersion
cyanobacterial
mosquito
same
Nodar- Lopez
unpub.
strains
logical features
or
were
purified
and
unpub.
that
of
or
from the
by
Thiery
characterized
data),
and
to
the
photomicrographs were performed as mentioned above
see" Sampling Methods"). The viable cell counts were

determined for the cell suspensions prior( a) larva feeding.
genus
The number of cells that survived in the larval guts and
and whose morpho-
the number( b) remaining in the water samples after the

60 minutes of the feeding experiment were also
belonging
Synechocystis
Synechococcus PCC 8905

will
site
Rippka (
Cells
data).
The
previously
breeding
and
Synechococcus
objective.
the
unicellular
strains
Synechocystis PCC 8906
determined.
The number of ingested cells ( c) was
be hereafter designated Synechococcus " type
calculated as the difference between the viable cell
Synechocystis" type 8906," respectively. The
counts determined for the cell suspensions prior to and
in
after the 60 minutes of larva feeding( Thiery et al. 1991).
CFU
This value was then converted to a percentage of ingested
8905"
or
abundance of the unicellular cyanobacterial strains
the water samples were estimated
by
counting
all
166
cells
100
for
x(
b-
to a-
each water sample
1- b/ a). The
The degree
c.
according to the formula:
number of digested cells(

of
digestion
d) is
equal
was estimated
from
the number of cells surviving in the larval guts at 60

minutes after the
beginning
of the experiment and was
ingested
expressed as a percentage of the
percentage of digested cells was calculated
the
formula: 100
b-
x( a-
The
cells.
according to
c/ a- b).
DECEMBER, 1993
during the same year-long study period, except in

December, and showed two peaks of high abundance
( 800 CFU/ml) in April and August, 1990.
Figure 2B presents the abundance of all the other
unicellular cyanobacteria found during the period of the

study. A strain belonging to the genus Synechococcus
with a cell size of 1. 3 m, which could thus correspond
to Synechococcus strain PCC 8975 isolated in June
1989, was present during the whole year in low abundance

RESULTS
with a peak value (
600
100 CFU/ml in June 1990)
occurring between the two peaks of Synechocystis" type

Population
Cyanobacteria
From
Dynamics
and
in
morphology
identical to those
PCC 8905
or
1991,
and
colonies of
pigmentation
Synechococcus
1990( 800 200 CFU/ ml), had the same size as the latter
but could easily be distinguished from it because it
of either
Figure 1
Synechococcus"
but was
peak
shows photomicrographs of
type
February to
8905"
density
from 850
nor in 1989 samplings of the same marsh, which belongs

to the genus Pleurocapsa, was discovered to be present
200 25 CFU/ ml during the
only in August and December 1990, and at a density
varied
from April to October, 1990(
season
150
contained phycoerythrin that gave the cells a blackish
pigmentation. Finally, a strain found neither in the 1988
10 CFU/ ml in December, 1990,
always greater than
larval
October, 1990, the second from February to October,

1990, and the third, which was only present in August
strain
each sample.
CFU/ ml in
8906." Three new Synechocystis strains were found in
Synechocystis strain PCC 8906 were found
the purified strains and of the new isolates. The

of
of
this breeding site. The first was present from June to
February 1990 to February
cyanobacteria whose
were
Abundance
and
Green Algae
2A). Synechocystis " type 8906"
was
also
Fig.
recovered
lower than 10 CFU/ ml( data
not shown).
Although the
total number of unicellular green-algae was ten times
tit
Ail
Figure 1.
'
fin
Yrr15WY
Photomicrographs of the axenic strains Synechococcus PCC 8905( 1) and Synechocystis PCC 8906( 3)
and of the respective homologous strains( 2) and( 4) present in the water samples collected from the
brackish
marshland
located
near
Fos/ mer( France)( bar represents 5
pm).
DECEMBER, 1993
167
1000
Synechococcus" type 8905"( 1 m)
A Synechocystis" type 8906"( 2.3 m)
CI
800-
4111k A
600-
400
200
2/ 90
4/ 90
6/ 90
8/ 90
10/ 90
12/ 90
2/ 91
Months
1500 -
Synechococcus"
type
8975"( 13 m)
Synechocystis( 2 m)
Q Synechocystis( 3- 4 m)
a+
gin
1000
41
Synechocystis PE+( 3- 4 m)
t
3
i
500
I.
2/ 90
4/ 90
6/ 90
8/ 90
11166.
10/ 90
12/ 90
2/ 91
Months
Mosquito Larva Season
Figure 2.
Population dynamics of unicellular cyanobacteria in the brackish marshland located near Fos/mer
France) for the year 1990 with emphasis during the mosquito larva season.( A) Synechococcus strain
type 8905" and Synechocystis strain" type 8906." ( B) Synechococcus strain" type 8975" and newly
found Synechocystis strains. CFU: colony forming units. When calculable, the density variation of
Synechococcus and Synechocystis
was
included between 9- 17
percent and
13- 53
percent,
respectively.
168
DECEMBER, 1993
higher than the total number of unicellular cyanobacteria,
The
six
different filamentous
1988 and 1989
date, in 1990 ( TABLE 1).

filamentous
strains
were recovered, on at
In
in cell
always found in the guts of both mosquito larva species
isolated in
one
addition,
belonging
cyanobacteria
Oscillatoria,( 4- 5
least
Different strains of unicellular green algae were
latter.
occurred at the same period than the
they
number of green algae available in the water sample
new
fluctuated from 65 to 93 and the degrees of digestion
genus
from 81 to 99 percent depending on the larva species and
one
to the
width) was present
in every
the water samples( data not shown).
sampling except in December. Although the cell density

of the filamentous cyanobacteria was not quantified, it
has been noticed that the new Oscillatoria
strain
strain
TABLE 1). The percentages of ingestion of the total
sampling
All of the unicellular cyanobacteria were recovered

in the larval guts of at least one of the two mosquito
strain and the
species tested. Cells of Synechocystis" type 8906" were
LPP group B 1, morphologically identical to the

PCC 8980, were the most abundant filamentous
found in the larval guts of the two mosquito species
during
cyanobacteria
the
year and
in
particular
throughout the year- long test period, while those of
Synechococcus" type 8905" were recovered only from
during
the larval season.
For
water samples collected from April to October 1990

the
comparison,
in the brackish
recovered
evaluated(
different
unicellular
TABLE 1).
marshland
At least
cell types were
green
algae
were
also
TABLE 2). Both species of mosquito larvae ingested

and
digested
cells of
Synechococcus" type 8905", Cx.
pipiens larvae ingested cells of Synechocystis" type
seven
morphologically
distinguished but none of them
precisely identified in the course of this study.

These photosynthetic eukaryotes were highly abundant,
8906" less efficiently than An. gambiae larvae, but the

degree of digestion by both species of mosquito larvae
was fairly similar.
ranging between, 1, 000 and 20,000

year except in December when
samples taken from April to October 1990 and the
were
density
their cell
CFU/ ml
Figure 3 depicts the initial cell density in the water
throughout the
they decreased
to only
40 CFU/ ml.
number of cells ingested by either An. gambiae or Cr.

pipiens
Characteristics
The
brackish
the
Breeding
Synechococcus " type
8905"
and
from 9 C in
unicellular cyanobacteria and for the total green- algal
population. These diagrams show that the two different
1990) to over 25 C in summer( June, August,
mosquito larva species did not preferentially ingest a

specific type of microorganism. Moreover, they show
These
5 C from min./ max.)
values were
Precipitation
station was recorded
month, except
in October
139
to
similar
very
to those measured at the
meteorological station.
increased
for
Synechocystis " type 8906," as well as for the other
winter
marshland water ranged
October 1990).
logical
Site
temperature fluctuations of the
seasonal
February
and
of
mm
and
and
that a single larva of An. gambiae ingested, within 60

minutes, ca. 3 to 4 percent( from April until October:
nearby
at the meteoro-
to be below 60 mm per
7915, 4313, 5717, and 2205 CFU) of the total
December 1990
population of unicellular cyanobacteria and green algae
112
mm
per
when
it
month,
a) or 3 to 4percent( idem: 555, 725, 829, and 778 CFU)
respectively. The depth of the marsh generally varied

from 5 to 30 cm near the edge but reached 50 cm in
of the total population of unicellular cyanobacteria( b)
October 1990
in December 1990.
Cx. pipiens ingested within 60 minutes roughly the
in Montpellier
same number of CFU of these two populations. Except

in October 1990, at least 1 to 3. 102 cells of either
Water
and
80 to 100
components
France)( 13). The
were
cm
analyzed
following
values
indicate the
range
1. 6- 27. 6 mg NH/ 1; 0- 0. 18 mg NO2-/ 1; 0. 12. 05 mg Cl-/1; 354- 708 mg HCO3/ l; 0 mg C032-/ I;

14 - 116 mg S042-11; pH 7. 4-7. 8; 33. 6- 62. 5 F for the
of variation:
present
in the
water samples(
Fig. 3). A single larva of
Synechococcus " type 8905" or Synechocystis " type
8906" were ingested in 60 minutes by either type of

larva.
total harshness and a water resistivity of 390 to 1130.

DISCUSSION
Ingestion and Digestion of Cyanobacteria and

Green Algae
by Mosquito Larvae.
ingestibility of the different filamentous
strains was recorded after the 60 minutes
feeding of An.
The
gambiae and
Cx.
Most
previously isolated ( Thiery et al. 1991) were also
the strains
recovered in 1990, and that the highest density of either
in the larval guts( TABLE 1),
except the
cyanobacteria or green algae was found between April
PCC 8971 producing phycowhich was never found in the guts of either one
and October, theperiod of the yearduring which mosquito
Pseudanabaena
of
larvae.
of
were recovered
erythrin,
pipiens
This investigation showed that strains whose
morphology and pigmentation are identical to those
strain
the two types of mosquito larvae.
larvae are the most abundant in the south of France. The

low abundance of cyanobacteria in December suggested
169
DECEMBER, 1993
Ii.0
3
g."...
e
t
k
m
k $
t
e
Q U
St
U U
c:,
CI)
U
1
t
t
0 \
k /
II
10
CI
2/
k
m
P.
41
7\
E
G.)
A
o
6..
es ]
t $
8
u
2 2
77aa ]
0
2faa /
2 -a
00
t 8/ ]
0
k
0K
c) /
@u
cu
k /
ka
2-
o _%
7..
170
DECEMBER, 1993
that some of the major factors influencing the density of

these photosynthetic microorganisms may be light
intensity and day length. In contrast, analyses of the

did
water samples
different
be
could
in the
not reveal much variation
correlated
the abundance
with
because it
allows comparison of
density
of the various
cited organisms.
This study
the
strain
PCC 8905
previously
selected on
properties and their

mosquito
Synechocystis
and
larvae,
Thiery
et al.
1991).
be
for
used
Indeed,
strains
light microscopy
200 CFU/ ml
over
be ingested
during
further is
diameter). This
240 and 1250 CFU/ ml
As
interesting
strain was present at a
o a
o
M
to study
in
and
in
density between
g Tr CD o
coo co g g
y`
8 8 8
M 8 8
o ao 0
S a
of
Q`
a previous report(
axenic cultures of
Synechococcus
Nov
Qa
c
larvae.
shown
c-
Z1
cell
digested by both types
oo
28
during the larval season and was
very efficiently ingested

mosquito
be
large Synechocystis ( 3 to 4
M oR rM N
6 2
and
this work and which might
the larval
digested by mosquito
larvae. Another cyanobacterium found in the course of
season and to
found to be
and pigmentation, were
density
present at a
8 C
identical morphological features, as determined by
with
VD 0000 0000
tin
r"
digestibility by
are good candidates to
mosquito control(
a a
a
PCC 8906,
strain
the bases of their physiological
ingestibility
a'
Synechococcus
and
&
a 2)
supported our previous conclusions that
two unicellular cyanobacteria,
A a'
tti
3
meaning
o
o
Viable counting is not an ideal way to enumerate

organisms because it might underestimate their
density,
recognized
y$
these
of
microorganisms throughout the year.
but it does have a significant, well-
la
71.
physico- chemical parameters measured that
Synechocystis
strain
Thiery et al.
PCC 8905
were
less
likely
and
1991),
PCC 8906
strain
.
o
to be
gNS
v
r
UU
ingested by An. gamb:ae than by Cx. pipiens larvae.

When
given natural water samples,
gambiae or
both
Cx.
pipiens
Synechocystis
Synechococcus strain"
show
ingested
in fact
no
strain
distinct
preference.
from 11 to 1
The
size of the
0
cd
m, which corresponds
by
larvae( Dadd 1971).
1a
0 0 0
0000
co
co
Synechococcus PCC 7942
of
gg&g
8
in the brackish
Preliminary experiments aimed at determining

toxicity
c,
and
to the size of the particles known to be ingested

mosquito
suggesting that larvae
cyanobacterial and the algal cells present

marshland ranged
of either An.
8906"
type
"
type 8905,"
larvae
similar quantities of
the
single
dose
of 2.4
of a population of second- instar
'
recombinants
within
24 hours,
whereas,
2.4
killed
larvae
ca.
of
95
Z.,
of Aedes aegypti
the same
Nicolas,
bioassay
are
digested
by
conditions (
in
larvae
release
B.
sphaericus
c", -
`''
personal
This indicates that cyanobacteria that
mosquito
killed
less than 10 percent of a population

communication).
i
00
percent
Cx. pipiens
x 104 cells/ larva
g.
containing the B. sphaericus toxin genes showed that a

x 102 cells/ larva
a
Ua
a^
E.,
aa
DECEMBER, 1993
Cell
density
in
Quantity ingested per larva after 60 min feeding
water sample
Anopheles
Month/ Year
4/90
Culex pipiens
gambiae
CFU/ ml
500
171
CFU/larva
CFU/ larva
800
200
200 _
x259
r 122
1 ^
21540
1540)
6/ 90
300 -
L'
120 ^
1300
7915
120
8/ 90
400
r
I
81';')
1120
4313
2013)
725)
13%
17%
800
160 -
v 4142
763)
, ; :!.
x( 18%
157
1320
1234
i
15100
108
IL
f
1 :"
6%
7%
1120
7137
417)
555)
5717
4*
4/j
2100)
829)
5551
715)
14%
14%
13%
a'
10/90
200
52
150
60
120
6315
2015)
2205
2160
778)
698)
35%
33%
14
r.
32%
12 green- algae
type 8905"
in " type 8906"
O other cyanobacteria
Figure 3.
Pie diagrams showing the relative cell density of the unicellular cyanobacteria and green algae in four
water samples collected at two-month intervals during the year 1990 compared with the number of cells
ingested per larva after the 60 min. feeding. aTotal number of cyanobacteria and green algae; bnumber
of unicellular cyanobacteria; cthe percentages represent 100 x( b/ a). The numbers of CFU/ml or CFU/
larva of either Synechococcus" type 8905" or Synechocystis" type 8906" are indicated above the arrows
pointing
out the
corresponding
sectors.
172
Culex, but
DECEMBER, 1993
less toxic
manuscript. This investigation was supported by a grant
towards Aedes larvae. This result correlates well with
from UNDP/World Bank/ WHO Special Programme
the previous observation that B. sphaericus toxins are
for Research and training in Tropical diseases( 880511)

and by the Institut Pasteur Paris.
toxins that act against
usually 100 to 1000 times less
are much
Ae.
efficient against
aegypti than against Cx. pipiens larvae( Davidson 1984,
Thiery
and
de Barjac 1989). In the
present
study, An.
REFERENCES CITED
gambiae or Cx. pipiens larvae ingested from 1x102 to

3x102 cells of Synechococcus strain" type
Synechocystis
type 8906"
strain"
during
8905"
and of
the season of
highest mosquito larva density except in October, 1990

when
these values dropped significantly.
Thus, if
as
mentioned above 2x102 cells of toxic cyanobacteria are
Angsuthanasombat, C.
S.
and
Panyim.
1989.
Biosynthesis of the 130- kilodalton mosquito

larvicide in the cyanobacterium Agmenellum
quadruplicatum PR- 6. Appl. Environ. Microbiol.
55: 2428- 2430.
sufficient to kill Cx. pipiens larvae, Synechococcus

strain "
type
8906"
could
brackish
8905,"
be
An.
for mosquito
In contrast,
in the
Chungjatupornchai, W.
1990.
mosquitocidal- protein
Expression of the
genes
of
Bacillus
thuringiensis subsp. israelensis and the herbicide-
known to be 10 times less efficient
resistance genebar in Synechocystis PCC 6803.
studied.
since
Cx. pipiens( Thiery,
gambiae than against
Current Microbiol. 21: 283- 288.
data), the abundance of cells of these two
unpub.
cyanobacterial strains would
An.
strain " type
control
B.
marshland
sphaericus toxins are

against
Synechocystis
and
used
likely
be insufficient for
gambiae.
In
Dadd, R. H. 1971. Effects of size and concentration of

particles on rates of ingestion of latex particulates
conclusion,
Synechocystis
and
strains
belonging
Synechococcus,
to the genera
and
in
particular
by mosquito larvae. Ann. Ent. Soc. Am. 64: 687692.
Synechocystis strain " type 8906" and Synechococcus

strain "
type
8905,"
cyanobacterial strains
appear
to
for cloning
be
and
appropriate
Davidson, E. W. 1984. Microbiology, pathology and
expressing the
Abundance of
genetics of Bacillus sphaericus: Biological aspects
encoding Bacillus toxins.

cyanobacteria is maximal during mosquito larval season
genes
which are important to field use. Mosq. News 44:

147- 152.
and cyanobacteria are well eaten although they are not
by
larvae among
Indeed, in
Davidson, E. W., M. Urbina, J. Payne, M. S. Mulla, H.

Darwazeh, H. T. Dulmage, and J. A. Correa. 1984.
Fate of Bacillus sphaericus 1593 and 2362 spores
used as larvicides in the aquatic environment. Appl.
Environ. Microbiol. 47: 125- 129.
view of using such recombinant cyanobacteria as

biological control agents, firstly it is necessary to
Gophen, M., and M. Gophen. 1986. Trophic relations
selected
mosquito
microorganisms present
in
water samples.
the
other
Since these
two essential points have been demonstrated, the next
step is the
reintroduction of
cyanobacteria
determine if
maintain
evaluate
in the
and
mosquito
for how
itself in the
the selected unicellular
breeding
long
mosquito
the minimum number
site.
can
between two agents ofsewage purification systems:
site and to
Algae and mosquito larvae. Agric. Wastes 15: 159-
a given species
breeding
of cells
necessary
to
be
168.
reintroduced, taking into account the fluctuation of the
density of the
targeted mosquito larvae.
Khawaled, K., M. S. Mulla,
and
A.
Zaritsky. 1989.
Distribution and abundance of algae in mosquito

development sites. Bull. Soc. Vector Ecol. 14: 71Acknowledgments
80.
We thank B. Gaven for technical
in
Marten, G. G. 1986. Mosquito control by plankton
collecting
Entente Interdepartementale de Ddmoustication du
management: the potential of ingestible green-
samples
and
G. Vigo for
assistance
water
analysis
algae. J. Trop. Med. Hyg. 89: 213- 222.
Littoral M6diterranden, Montpellier). Thanks are due to

R. Rippka for the determination
present
in the
and to P.
of the cyanobacteria
water sample collected
in
February
1990
Brey fer providing An. gambiae larvae. We are

indebted
to A. Bely for critical reading of the
greatly
Ohana, B., J. Margalit, and Z. Barak. 1987. Fate of
Bacillus thuringiensis subsp. israelensis under

simulated
field
conditions.
Microbiol. 53: 828-831.
Appl. Environ.
DECEMBER, 1993
M. Herdman,
Generic
assignments,
1987. Expression of the larvicidal gene of Bacillus
and properties of pure cultures of
sphaericus 1593M in the cyanobacteriumAnacystis
J. Gen. Microbiol. 111: 1- 61.
nidulans R2. Mol. Gen. Genet. 209: 396-398.
R. Y. Stanier.
strain
histories
cyanobacteria.
chimique
de l'
Dunod. Paris,
1979.
L' analyse
1966.
Rodier, J.
Tandeau de Marsac, N., F. de la Torre, and J. Szulmajster.
Waterbury,
Rippka, R., J. Deruelles, J. B.

and
173
eau.
Eaux
chimique
et physico-
naturelles- eaux
usees.
Thiery, I. and H. de Barjac. 1989. Selection of the most

potent Bacillus sphaericus strains based on activity
ratios determined on three mosquito species. Appl.
412pp.
Microbiol. Biotechnol. 31: 577-581.
Silapanuntakul, S., S. Pantuwatana, A. Bhumiratana,

and
1593
comparative
Thiery, I., L. Nicolas, R. Rippka, and N. Tandeau de
of Bacillus sphaericus strain
Marsac. 1991. Selection of cyanobacteria isolated
H- 14
from mosquito breeding sites as a potential food
K. Charoensiri.
persistence of
and
against
toxicity
Bacillus
mosquito
environments.
1983. The
thuringiensis
serotype
larvae indifferent kinds
J. Invert. Pathol. 42: 387- 392.
of
source for mosquito larvae. Appl. Environ.

Microbiol. 57: 1354- 1359.
DECEMBER, 1993
WATER DEPTH AND LARVAL DENSITY AFFECT DEVELOPMENT

AND ACCUMULATION OF RESERVES IN LABORATORY
POPULATIONS OF MOSQUITOES'
S. E. Timmermann2 and H. Briegel2
ABSTRACT: The larval development ofAedes aegypti, Anopheles albimanus, Anopheles gambiae, andAnopheles
stephensi were investigated under two different rearing protocols: increasing water depth( 0.5- 14 cm) or increasing
larval densities( 100-> 1500 L/pan) under otherwise constant conditions( 27 C, 14: 10 L:D). The times of pupation
and eclosion were recorded daily, and the protein and lipid reserves were analyzed after eclosion. Defined by the total
eclosion of males and females, the highest larval survival occurred at an optimal water depth of between 0.5 and 1
cm. At greater water depths eclosion was reduced. Anopheles usually did not survive in water deeper than 2 cm,
although dissolved food could extend the maximum depth that would allow survival up to 5 cm for An. albimanus
and An. gambiae. Eclosion of Ae. aegypti was affected less by increasing water levels, in that 50 percent of the
population still eclosed in 14 cm. Larval density strongly affected development, as evidenced by longer duration and
reduced body size. To allow intra- and interspecific comparisons, all caloric data of teneral imagoes were converted
to " sizespecific caloric contents."
These values exhibited the synthetic potential for total protein and lipid,
irrespective of actual body size. Aedes aegypti differed from Anopheles. Protein synthesis ofAe. aegypti reached a
fairly constant level at the end of the larval period, regardless of the rearing conditions, whereas the lipid synthesis
increased exponentially when regressed against adult body size. In Anopheles, the corresponding relationships were
consistently on a lower level than in Ae. aegypti despite their overlap in body size. When combining the sizespecific
caloric contents of protein and lipid for all larval treatments of Anopheles, their regressions against body size had
identical slopes.
INTRODUCTION
subsequently are required for various energy demanding

behavior patterns, and for reproductive and metabolic
processes( Briegel 1990a, 1990b). Larval development
Variability in body size is observed in all species of

the animal kingdom, both wild and domesticated, but its
influence
on
is largely determined by uneven food distribution and

the resulting intraspecific competition for these limited
resources, which ultimately is reflected in variable body
the physiological level has been analyzed
in only limited numbers of species,
primarily vertebrates
Schmidt- Nielsn 1984). The extent of variation within
size( Mori 1979; Fish 1985; Bradshaw and Holzapfel
the genetic constitution of a given species often appears
1992). Body size, as an expression of biomass, has been
to cover largely one- third of the maximal size,

excluding
reported as pupal life weight( Gilpin and McClelland
endocrine
or
developmental
represent suitable models

of variable
body
size
Insects
1979; Slansky 1982; Hawley 1985) or as the cube of a
for studying causes and effects
morphometric parameter ( Briegel 1990a, 1990b) as
for
abnormalities.
several reasons.
First,
their
suggested by Schmidt-Nielsen ( 1984). In mosquitoes,
relatively short generation times reduce the time required
with their medical importance and the worldwide efforts

to control their tremendous reproductive potential, a
for experiments and their small size allows environmental
factors to be
increases
Second,
manipulated.
occur
after
adult
consequence, the size attained
no
eclosion.
by
further
Third,
size
as
the final molt reflects
the synthetic processes and their limitations through

environmental conditions
In
mosquitoes, the
providing
during
larval
sufficient reserves
the larval period.
is
number of reports deal with aspects of variable body

size and fecundity in relation to transmission potential
Gillies and De Meillon 1968; Haramis 1985; Hawley

1985; Nasci 1987; Van Handel and Day 1989; Grimstad
and Walker 1991).
for
We have undertaken a detailed analysis of imaginal
for the teneral imago that
caloric reserves, reproductive potential, and metabolic
period
responsible
In honor of Professor Arden O. Lea

2lnstitute
of
Zoology, University
of
Zurich, CH- 8057 Zurich, SWI' I ERLAND.
function
parameters as a
175
DECEMBER, 1993
done for Aedes
of
responses to larval stress on survival, developmental
body size. This analysis was
Briegel 1990a)
aegypti (
and
for four
time, body size, and teneral reserves.
Anopheles species( Briegel 1990b) by varying the larval

from
conditions
extreme
and starvation to
crowding
optimal rearing conditions with a maximal food supply.

These treatments
between
female
body
was
the total
in Ae.
protein of
linearly
were
body
with
in
size
aegypti.
this study: Aedes aegypti( L.) UGAL strain from Georgia,

USA;
Under
size
strain of Anopheles ( Cellia) stephensi Liston. They
were colonized continuously for more than 15 years in

our laboratory at 27 C, and kept at a relative humidity
body
several culicines,
but in the
of 85 5 percent under long day conditions( 14L: 10D)
lipid were considerably

lower and for both components linear correlations were
Briegel 1990b). Furthermore,
with a 20 min. simulated sunset and sunrise. Adults

were maintained on 10 percent sucrose and fed twice a
body
week on consecutive days on a guinea-pig for 20 min.

Newly hatched larvae were counted into the rearing
size did influence the size of the blood meal and therefore
fecundity
the
ingested
ones
of
female Ae.
the
blood
pans. This number is referred to as 100 percent of the

population.
Aedes aegypti and An. stephensi were
volume of small
eggs
of mature
number
Large females
aegypti.
more than twice the
and
albimanus
showed an exponential
anophelines, teneral protein and
most appropriate (
Anopheles ( Nyssorhynchus)
Wiedemann, a Californian strain; Anopheles ( Cellia)

gambiae Giles, strain 16c55 from Nigeria; and an Indian
all
eclosed males
newly
correlated with
Briegel 1990a). Total lipids
increase
We have used the following species and strains in
size to the
The relationship
teneral caloric contents per
close
particularly
females
and
would allow.
and
size
circumstances,
body
allowed us to push
the genome
extremes that
was
almost
et al.
grown in pans of 19. 5 x 29 x 5 cm, while An. gambiae
1956; Ja1i11974; Hien 1976; Steinwascher 1982; Haramis
was grown in pans of 16 x 24 x 5. 5 cm containing
1985; Hawley 1985; Briegel 1990a; Bradshaw
and
distilled water 1cm deep, i.e. 450or300 ml, respectively.
Holzapfel 1992). In Anopheles, the comparatively low

caloric content of teneral females was hypothesized as
The larvae were fed our standardized food mixture of
quadrupled(
Colless
Chellapah 1960; Woke
and
for evolving different reproductive strategies

than known from the Culicini ( Briegel and HOrler,
a reason
For the
laboratory
colonization
optimal and standardized

attempted
in
rearing
order to obtain
mosquitoes,
of
homogenous
size(
Lea 1963;
Gerberg 1970;
for this investigation. To assure standardized feeding,
usually
we used a set ofspoons with defined volumes in multiples
populations
of approximately 36 mg; caloric contents and quantitative

details are given by Briegel( 1990a).
conditions are
with synchronous pupation and eclosion and uniform
Briegel 1990a,
In our experimental setups for larvae we used rat
data. In the present report two parameters
containers of macrolon ( 24 x 40 x 15 cm), allowing

maximal water levels of 14 cm. Food was added by
the larval milieu that fulfill the routine conditions
evenly sprinkling the powder over the water surface.
1990b). This is
physiological
of
1: 1: 1). The daily and total food supply was established

empirically and optimized over years for each species
colonized( TABLE 1). These schemes were the standard
1993).
imaginal
powdered rat chow, lactalbumin, and brewers yeast

(
a prerequisite
for obtaining
density
Under suboptimal conditions, which prolonged larval
The
types in the
development, the feeding period was extended by adding

daily the last spoon unit until pupation was completed.
our emphasis was rather the
In general the amount of food was kept constant per
were varied: the depth of the water and the
identical
under otherwise
Dahl( 1988), but
efficiency in food
different
acquisition,
caloric reserve accumulation as
imaginal
revealed
such
populations,
mortality in
as
feeding
ultimately
responses
container, and not per individual larva, irrespective of
in
different water volumes.
already had
In all experimental arrangements, pupae were
of mosquito
collected daily and transferred into small glass vials for

recording the individual eclosion times. In this way, the
Wada 1965), decreased larval
time of pupation and of eclosion, and the number of
1984),
Moore
fecundity (
reflected
of
and
of
development
pupation and eclosion rates(

et al.
in terms
assessed
experiments
duration
all stages (
survivorship,
1972; Reisen
and
Previous
body
densitydependent
size.
larval
and constant conditions.
species selected represent
sense of
meaningful
Barbosaet
oviposition, adult
and
al.
longevity,
Fisher 1969; Fish
and
Carpenter 1982; Livdahl 1982; Steinwascher 1982;
Hawley 1985;
Briegel 1990a, 1990b; Walker
1991; Kitthawee
performed
to
et al.
1992). Our
measure
et al.
experiments were
individual compensatory
survivors were established. Shortly after eclosion,

imagoes were sexed, their wing length measured, and its
cubic value(= WL3) used as an estimate of body size.
Subsequently, they were fixed individually in a drop of
hot ethanol ( 90 C for 3 min. in 10 x 70 mm reacting
tubes) for biochemical analysis.
The
analytical procedures
for caloric
contents were
176
TABLE 1.
DECEMBER, 1993
Feeding regime under standardized conditions for four mosquito species. Spoons
with an average content of 35. 6 0.8 mg and multiples thereof were given in daily
succession. The total amount of the food mixture( mg), added until completion of
development, is converted into caloric input, and expressed per larva.
Aedes
Anopheles
Anopheles
Anopheles
aegypti
albimanus
gambiae
stephensi
Spoon/ day
10
4
4
Total
weights(
Total input(
948
689
1286
544
2938
2137
3987
1687
mg)
cal)
Total caloric
input
per
larva
14.7
7. 1
identical to our previously used methods( Briege11990a,

1990b). Total protein, lipids, and carbohydrates were
measured
for individual
caloric values
based
on
were then related
5. 6
with all other parameters until eclosion. The days of
pupation and eclosion were recorded and newly eclosed
mosquitoes and converted to
imagoes were counted, their wing length measured and
the relation of 4 cal
subsequently analyzed for total protein, lipids, and

carbohydrates. Data for males were gathered and
protein or carbohydrate, and
data
8. 0
to
cubic value of wing length.
cal per
body
per
mg
mg lipid. These
size, estimated
by
the
To allow direct comparisons
compiled in TABLES 2 and 4, but were given only little

consideration in this report.
of size and caloric contents, between different species,

all caloric
data were multiplied by the factor of 10/ WL3.
This factor
application
represents an uniform
led to the"
body
size and
its
sizespecific caloric content per
mosquito."
Two
experiments were carried out
four
mosquito
water
depth
was
density
In the
kept
identical
0. 5 to 1 cm for Anopheles and Ae. aegypti as well. The
for the
sex ratio was always close to 1: 1, males being slightly

more frequent, never exceeding the females by more
series,
gradually increased from 0.5
until eclosion.
was
an
In the first
cm, while all other parameters were
hatching
in
species except
manipulation of two parameters.
When reared with larval food sprinkled on the

water surface, the highest larval survival of both sexes
in three species varied between 80-95 percent of the
larval counts, whereas in An. gambiae it was only 72

percent. Optimal survival, defined by total eclosion of
males and females, was consistently within depths of
RESULTS
manner with
Effect of Water Depth on Larval Survivorship
the
cm
to 14
than 10 percent in our experiments. Detailed results for
constant
from
total eclosion of the whole population and the single day
second series the
gradually increased from 100
larval
per pan to
1500 or more, keeping the water level constant together
with maximal eclosion of females are given in TABLE

2.
In general, the total percentage of a population

adulthood decreased with
increasing water
reaching
DECEMBER, 1993
TABLE 2.
177
Effect of water depth( cm) and two modes of feeding on the percentage of eclosion. Total
females
eclosion of males and

addition,
for females, the
day
of the larval counts. In
expressed as percentage (%)
of maximal eclosion(%)
is given.
Modes of feeding
Food
Water
Total
Female
Total
Female
depth
eclosion
eclosion
eclosion
eclosion
day
cm)
Aedes
aegypti
200 larvae)
Anopheles
albimanus
300 larvae)
Anopheles
gambiae
500 larvae)
Anopheles
Food dissolved
sprinkled
stephensi
300 larvae)
day
0.5
49
10
58
89
85
88
75
88
78
82
10
53
93
54
40
11
42
84
11
48
10
59
12
42
76
11
64
14
54
12
48
74
12
41
0. 5
95
10
87
80
11
83
70
11
66
10
17
36
19
16
43
18
42
0. 5
72
10
37
10
38
36
11
57
34
65
13
41
59
14
45
22
15
41
0. 5
89
10
47
92
81
70
11
84
depth. In Anopheles none of the first and second- instars,
Ae. aegypti for all water levels, e.g. from 49 to 89
respectively, survived in more than 2 cm. Furthermore,

TABLE 2 shows the time required for pupation of the
percent at 0. 5 cm or, from 54 to 74 percent at 14 cm.
females and thus delineates the duration
the decrease at 14 cm. Anopheles were affected in
major part of the

of their
which
larval
larval
and pupal
development. At
depths in
several ways by this feeding regime. In An. albimanus
developmental
andAn. gambiae, dissolving the food allowed successful
females
development down to 5 cm water depth, whereas in An.
1 to
a different
stephensi it failed completely ( TABLE 2). Under all

circumstances, i.e., irrespective ofthe feeding conditions,
water depth, larval density or sex, the pupal period was
Following conventional
consistently 1. 5 to 2 days, only males pupating a little
survival was possible, the
all
period until eclosion of the major segment of

was extended
by only
3 days in Ae.
aegypti.
1 to 2 days in Anopheles
The same experiments were repeated with

mode of larval food application.
routine in
Female eclosion increased correspondingly, except for
and
laboratory colonies, food of the same amounts
earlier than females.
as before was first dissolved in a negligible volume of

distilled water and then
added to the
way, an optically homogenous
rearing pans. In this
body
Effect of Water Depth on Caloric Content
was obtained
The same material was analyzed for the caloric
with
the foe6lequally distributed throughout, minimizing

floating at the surface. The data in TABLE 2 reveal a
content of female reserves shortly after eclosion. In all
considerably higher survival of the larval population for
percent of the means when grown at
water
species tested, female body size varied within 10 to 20

different
water
DECEMBER, 1993
178
depths; Ae. aegypti grew larger in deeper water( ANOVA
In Anopheles the situation was more complicated
F= 8. 59; df= 5, 400; p= 0. 0001), reaching the maximum

at 14 cm. In Anopheles maximal sizes were observed at
because the three species behaved differently. The

survival ofAn. albimanus in deeper water by dissolving
the food was accompanied by reduced caloric protein
and lipid contents of the newly eclosed female. An.
depths
of
2 cm,
or
whereas
More informative
quantitative analyses of the
lipids,
and carbohydrates.
0. 5
the minima were at
results
were
gained
females for their
In newly
cm.
from the
protein,
females the
eclosed
gambiae grown in deeper water showed an increase
mainly in lipid reserves( TABLE 3).
carbohydrates( mainly glycogen and trehalose) always

contributed
less than 10 percent of the total
therefore we focused
our
interests
calories and
lipids( TABLE 3).

In Ae.
depths
defined an optimal water depth for each species. At this
showed a similar variation with
water level the larval density was varied from 100 larvae
Mean lipids however, had clearly

with the food dissolved( TABLE 3).
per rearing pan to extreme crowding conditions, causing
in Ae. aegypti applying the food in dissolved
food shortage. This in turn affected their developmental
aegypti, grown at
feeding
increased
form
increasing
water
modes.
values
Obviously,
Based on the highest percentage of total eclosion of
males and females in the preceding experiments, we
mean protein values
both
Effect of Larval Density on Survivorship
on total proteins and
success.
was superior.
TABLE 3.
arearing stress which was accompanied by apronounced
Effect of water depth( cm) with standard densities( larvae/pan) on the caloric content of total protein
P) and lipids( L) of eclosing females of four mosquito species. Means S. E., N= number of females,
at
least
a third of which was analyzed
for each
reserve component,
Caloric content
Water Depth
cm)
Ae.
aegypti
200 larvae)
An.
albimanus
300 larvae)
An.
gambiae
500 larvae)
An.
stephensi
300 larvae)
Same
amounts of
E= the sum of protein plus lipids.
P*
L*
E*
0. 5
43
1. 59 0. 15
1. 61 0. 15
0.75 0.27
1. 50 0.29
2. 3
3. 1
93
1. 83 0.20
1. 700. 15
1. 31 0.21
1. 46 0. 32
3. 1
3. 2
93
1. 69 0. 17
1. 71 0. 15
1. 49 0.27
1. 84 0.35
3. 2
3. 5
69
1. 52 0. 15
1. 61 0. 19
1. 05 0.25
1. 85 0.27
2. 6
3. 5
10
56
1. 65 0.23
1. 660. 16
1. 03 0. 22
1. 66 0. 33
2. 7
3. 3
14
52
1. 84 0. 23
1. 56 0.30
1. 23 0.26
1. 75 0. 30
3. 1
3. 3
0. 5
98
1. 12 0. 16
0. 34 0. 10
1. 5
83
1. 14 0. 14
0.45 0. 11
1. 6
70
0.960. 10
0.52 0. 12
1. 5
2*
14
0. 63 0.07
0.29 0.08
3*
23
0.84 0. 10
0. 34 0.09
1. 2
5*
12
0.70 0.28
0. 240. 03
0.9
0. 5
94
0. 63 0. 14
0. 18 0.04
0.8
64
0. 71 0. 10
0. 16 0. 04
0.9
76
0.81 0. 13
0. 16 0. 04
1. 0
2*
170
0.83 0. 13
0.38 0. 19
1. 2
3*
128
0.88 0. 11
0.29 0.07
1. 2
5*
49
0.87 0.07
0.260.05
1. 2
0. 5
125
0.76 0. 12
0. 17 0. 04
0. 9
137
0.67 0. 14
0. 14 0.02
0.8
97
0.75 0. 12
0. 17 0. 04
0.9
food, but dissolved in 5
ml
distilled
water.
0.9
At
of the
of Ae. aegypti,
densities
all
larval
indicative
Anopheles
density- tolerant
severe reduction
metamorphosis,
The minimal developmental time for larvae was

always seven ( Ae. aegypti) or eight days ( Anopheles)
TABLE 4). In
but under crowding, the larval periods were extended up
percent or more
species(
increasing
as a consequence of
there was a
79
completed
population
of a
179
DECEMBER, 1993
in the
densities,
total number of
to three times, and eclosion was postponed up to one
month after hatching.
larvae reaching the imaginal stage. As the number ofAn.

albimanus
larvae
developmental
69
were
11
percent
in An.
from 91
doubled
from 81
success was reduced
percent, and
a reduction
increased from 100 to 600, their
was
these densities
stephensi
percent
once more,
to 59
and to zero
to
caused
If densities
percent.
developmental
in An. stephensi
percent
success
fell to
in An. albimanus.
of
developmental
larval
success
An. gambiae might be

species
in Ae.
there was a trend
for
the larval
aegypti,
density
An.
was
too
low,
albimanus, or
percent ( TABLE
5). In all Anopheles
The quantitative analyses of the caloric contents of
eclosing females are summarized in TABLE 5 for each

species and density. The total caloric content changed in
stephensi.
reduced survival
relation to body size and protein always prevailed over
100 L/ pan
the lipids. On a percent basis, in Ae. aegypti the protein
such as
An.
47
density- tolerant
An.
or
556.52; df= 4, 947; p= 0.0001), the maximal reduction

of size was
reduced
percent.
considered amore
albimanus
than
Alternatively,
when
An.
only
44
to
Increasing densities caused significantly reduced
body size of all female Ae. aegypti ( ANOVA F =
Therefore,
100 to 1500)
density (
Content
species tested, similar reductions were observed( 35- 46

percent), with significant values of 0.0001( ANOVA).
Anopheles gambiae had a different response in that a 15fold increase
Effect of Larval Density on Body Size and Caloric
gambiae.
decreased between maximal and minimal caloric
Effect of larval density( larvae/pan) on the percentage of eclosion. Total eclosion
TABLE 4.
of males and
females expressed as
for females, the
day
percentage(%)
of maximal eclosion(%)
of the larval counts. In addition,
is given.
Eclosion
Larval
density
day
larvae/ pan)
Aedes
1
aegypti
cm)
Anopheles
0. 5
albimanus
cm)
Females
Total
100
79
67
200
93
61
400
82
74
800
81
40
1000
83
11
31
100
81
11
68
300
87
10
50
600
69
13
33
100
73
10
46
500
78
11
51
1000
71
14
29
1500
44
15
28
100
91
55
300
90
10
33
600
59
15
39
1200
11
18
27
11
18
16
1200
Anopheles
0. 5
cm)
Anopheles
gambiae
stephensi
cm)
extremely
crowded
180
by - 56
contents
percent, and -
83
percent
for lipid. In
sizespecific protein and sizespecific lipid contents
Anopheles, this decrease was- 72 percent for protein and

77
percent
for lipids in An.

79
protein andpercent
for
percent
for
same regression line, irrespective of species or food
application. This indicates that total protein synthesis at
albimanus,-
82
percent
larval lipid
its
percent
for lipids in An.

is
synthesis
sensitive to crowding conditions than the

of total protein, with
regressed against body size. All protein values fit the
for lipids in An. gambiae, - 64
protein and -
Obviously,
stephensi.
45
the end of the larval period was the same for all species
more
and treatments, and was strictly size- related ( Fig. 1).
larval synthesis
Lipid synthesis behaved similarly, also following a
more structural nature.
significant linear regression of identical slope, but with
less than half the efficiency, indicated by a considerably
Size- Specific Caloric Contents of Female Mosquitoes
lower intercept. The total carbohydrates contributed
To compare more conclusively caloric data from

different individuals, species and treatments, we
introduced the concept of the " sizespecific caloric
content" of protein or
only a minor caloric segment without any correlation.
A different situation was revealed for Ae. aegypti.

Their sizespecific protein content remained more or
lipid per insect. For that purpose,
less constant under all rearing conditions, while their
the means from experimental female cohorts presented

in the previous TABLES were recalculated( see Material
and
Methods)
body
size (
sizespecific lipid content showed a positive allometric
increase with body size( Fig. 2). More lipid was deposited
and the values obtained plotted against
Fig. 1).
For
all
Anopheles
species
per size unit, as a female larva grew larger; this was also
tested,
true for females originating from different modes of
there were significant linear correlations between both,
TABLE 5.
DECEMBER, 1993
larval feeding.
Effect of increasing larval density( larvae/pan) at optimal water levels( cm) on the body size( WL3) of
eclosing females and on the caloric content of total protein ( P) and lipids ( L) at eclosion of four
mosquito species.
reserves,
Means S. E., N=
number of females, the majority of which was analyzed for
I=the sum of protein plus lipids.

Larval
density
larvae/ pan)
Aedes
1
aegypti
cm)
Anopheles
0. 5
albimanus
cm)
Body
WL3
size
Caloric Content
N
X
3. 4
100
37. 604.40
42
1. 88 0. 10
1. 49 0. 23
200
35. 05 2. 99
95
1. 84 0. 17
1. 32 0.21
3. 2
400
29. 58 3. 69
160
1. 41 0. 21
0. 87 0. 19
2. 3
800
19. 91 4.03
283
0.82 0. 28
0. 35 0. 14
1. 2
1000
21. 53 3. 70
372
0. 97 0.28
0.260.09
1. 2
2. 4
100
46. 53 3. 51
37
1. 760. 15
0.65 0.09
300
34.52 3. 58
130
0.91 0. 10
0. 26 0.05
1. 2
600
24.90 4.75
195
0.49 0. 11
0. 15 0. 02
0.6
1200
0
0
Anopheles
0. 5
Anopheles
1
gambiae
cm)
stephensi
cm)
extremely
crowded
100
34.43 3. 43
41
0.940. 09
0. 48 0. 07
1. 4
500
31. 26 4. 25
138
0.78 0. 14
0.21 0. 06
1. 0
1000
23. 993. 68
171
0.62 0. 18
0. 10 0.02
0. 7
1500
22.29 3. 65
192
0. 52 0. 16
0. 10 0.02
0.6
1. 8
100
36. 24 2. 81
42
1. 17 0. 07
0. 61 0.06
300
27. 77 2. 73
123
0.73 0. 05
0.36 0.05
1. 1
600
21. 87 2. 94
183
0.49 0.09
0.09 0. 02
0.6
1200
20.37 2. 33
66
0.49 0. 10
0. 10 0. 01
0.6
19. 84 2. 68
47
0.42 0.06
0. 11 0.03
0. 5
181
DECEMBER, 1993
0.400
0.300 --
0)
I
w
ig.
11
Go
0.200 --
V
U
V,
U
G.
C/ 1
6
cn
0. 100 --
0-
ti
3. 0
2. 5
50
40
30
20
10
3. 5
I
mm
Body Size
Figure 1.
Size- specific caloric protein( P) and lipid( L) contents per teneral female in relation to body size of all
three Anopheles tested( abscissa: cubic value of wing length above, and linear dimensions in mm below).
Each point is the mean from one experimental cohort of females being raised at variable water depths or
larval densities. Data from TABLES 3 and 5 were recalculated to sizespecific caloric contents by using
3),
the formula ( cal x 10 x WL-
respectively. The corresponding linear regressions are the following:
Y=0.004X+0. 132( N=21, r=0. 824, p< 0.001) forprotein, and Y=0.004X-0.049( N=21, r=0.777, p<0.001)
for lipids. Total carbohydrates showed no significant correlations with size, therefore only their average
is given( 0.035 0.016, N=21, open square). Symbols are: filled triangles, An. albimanus; filled circles,
An.
gambiae;
filled
squares,
An.
stephensi.
182
0. 600
DECEMBER, 1993
E
0
0.400
a
l
0
U
o
i
U
o
0.200-r
20
30
I
2. 5
20
3. 5
50
1
3. 0
40
'
30
1
2. 5
I
3. 0
40
I
50
1
3. 5
If
mm
Body Size
Figure 2. Size- specific caloric protein( P) and lipid( L) contents per teneral female ofAedes aegypti in relation to
body size( abscissa: cubic value of wing length above and linear dimensions in mm below). Each point
is the mean for one experimental cohort of females being raised at variable water depths or larval
densities, or with different modes of feeding. Data from TABLES 3 and 5 were recalculated to sizespecific caloric contents
by using
the formula (
cal
10
WL-3),
respectively. The allometric
exponential) regression for lipids is Y=0.002X1. 498( N=23, r=0.700). The linear regression for protein
is Y=0.417+ 0. 001X ( N=23,
r= 0.01, n. s.).
Because protein and carbohydrates showed no significant

correlations with size, their average contents are presented: protein 0.4200.049( N=23, closed square)
and carbohydrates 0.0570.021( N=23, open square). Open circles represent lipid values when food was
dissolved.
In
an attempt to
normalized the
further
for the corresponding

structural components).
revealed
eclosed
by
generalize our
lipid data( mainly

protein
data,
we
the data revealed two characteristics: first, larval
reserve components)
lipogenesis was linearly correlated with body size in
content (
The larval lipid
primarily
synthesis, as
the mean sizespecific content in

newly
females, therefore
calorie of protein present (
was
calculated
Fig. 3).
per
one
This treatment
of
Anopheles but in an allometric fashion in Ae. aegypti.

Second, the range of larval lipogenesis covered a
much lower level in Anopheles( 0.2- 0.5 cal lipid/ cal
protein) than in Ae. aegypti ( 0.2- 2.0 cal lipid/ cal
protein).
DECEMBER, 1993
183
Aedes aegypti
2. 000
l
0
r$
1. 000
a.
U
o
o.
10
20
30
40
50
U
U
6
r"
Anopheles
0.600
0.400
0. 200
0
20
1
10
30
1
2. 5
40
1
3. 0
50
1
3. 5
mm
Body Size
Figure 3. Demonstration of increasing larval lipogenesis in relation to protein synthesis, and dependence on body
size for both in Aedes aegypti and three species of Anopheles combined. The sizespecific lipid contents
showed in Figure 2, were normalized per unit protein, expressing the lipid per calorie of protein present.
When plotted against body size, a positive allometric correlation was found for Aedes aegypti
Y=0,004X1. 476, N=23, r=0.608) and a linear correlation for the Anopheles( Y=0.011X-0.027, N=21,
r=0.669, p<0. 01). Each point represents the mean of an experimental cohort.
DISCUSSION
newly eclosed females. All results were related to the
body size of the teneral female mosquito. When

The biosynthetic
potential of mosquito
larvae has
been studied by measuring the content of total lipids
and
carbohydrates, the main reserve components, and total

protein as
primarily
of structural
function,
present
in the
comparing the data after rearing with increased levels of

water or larval densities, Ae. aegypti was found to differ
in several fundamental aspects from the anophelines.

In essence, Anopheles larvae
of all species tested
the same
showed
and
lipids:
proportions
in the
synthesis of protein
the
protein always prevailed over
total (
1988) had also recognized a nutritional advantage of
Ae. aegypti larvae when expressing the lipids per unit
lipids, and
consistently were below 10 percent of the

This was particularly interesting in
to Ae. aegypti, which are of similar size but
protein.
carbohydrates
Depending on environmental conditions ( density
1).
Fig.
comparison
had
DECEMBER, 1993
184
a greater protein content and a much
and
depth), the larval period showed a considerable
flexibility in its duration as opposed to the constant
higher lipid
was a
duration of the pupal period. This accentuates the
between larval
importance of the phagoperiod, suggesting certain
findings
thresholds to be reached before pupation is initiated, as
other
wing length and protein were equally found in
species ( Van Handel and Day 1989; Briegel, unpubl.
already recognized by Gilpin and McClelland( 1979).

Larval mortality increased with increasing water depth
and/or larval density, contrary to Hawley ( 1985), and
for lipids
therefore the concept of a critical size or caloric threshold
Aedes
is even more compelling. Similar to Manduca( Nijhout

1975), the larva appears firstly determined to feed and
in their largest individuals. The latter
content
of
result
an
exponential
lipogenesis and body
correlation
our earlier
size,
supporting
Strictly linear correlations between
Briegel 1990a).
research) and positive allometric correlations
in Aedes
also revealed
were
triseriatus(
Briegel,
unpubl. research).
believed to be
ships are
mosquitoes.
of general
field- collected
and
Van Handel ( 1986)
culicine
synthesize in order to reach its critical threshold( s) as
maintained
rapidly as possible. Such thresholds were of a
validity in
best be
morphometric nature in Manduca( ibid.) andAe. aegypti
by Day
described
mosquitoes, as
can
relation-
explained
as a
Gilpin and McClelland 1979), whereas Chambers and
Klowden ( 1990) claimed the glycogen reserves to be
ultimately reflecting the larval food
of size,
supply.
In this
These
Differencesbetweenlaboratory-
and
matter
and
albopictus
critical. At any rate, an urgent need to feed and grow also

introduced the
governs the larval period of all Culicidae, but to a
term of" sizespecific caloric contents" ofnewlyeclosed
different degree among the members of the two
to be a most conclusive ex-
subfamilies. A further aspect, and possible source of
imagoes. This
proved
allowing the
pression,
results.
have defined
report we
Having
i. e.,
conditions,
findings. Since in
with
represent a
the
larval
In Ae.
content was
level
of
affected
protein
nearly
by
however, the
cal (
duration of larval development, and consequently in the

extentoflarval biosynthesisbecauseasubstantial feeding
to
period might be inserted. Once these critical conditions
appear
adult
have been met, pupal development being confined to a
during
closed system is initiated. It is programmed for
efficiencies.
autonomous expression, within a constant, species-
protein
specific duration of 1. 5 to 2thys, independent ofprevious
biomass
identical
sizespecific
fairly
and remained at a
Fig.
2).
it
Obviously,
in
was
barely
conditions proceeded
content of
the preference of mosquito larvae for shallow water
followed the same exponential regression,
pools under natural conditions, which are routinely
of the
The
larval
sizespecific
conditions, and at a much
Anopheles
water bodies are accessible to mosquito larvae as far as
was all
they are physically structured by leaves, roots, grasses,
our
feeding strategies of female

anophelines, as reflected by their multiple feeding
behavior as a means of improving their teneral reserves
about
different
Hurler 1993, Klowden
and
encountered in field situations. In other words, deep
the
Fig. 3). Therefore,
sizes of Aedes and
body
lipid
the more remarkable. These findings support
and
1965).
The surprisingly low water level of 0. 5- 1 cm for
all
higher level than in Anopheles(
Briegel
optimal development for all species tested might explain
under
maximal rates.
hypothesis
larval conditions as already recognized by Wada
constant
at
allAe. aegypti
similarity in
the
the larval treatments, arguing that larval
synthesis
irrespective
system is the question of temporal gates. If such gates do

exist, as in other insects( Truman and Riddiford 1974),
then one could expect considerable variations in the
to their
same positive cor-
the anophelines
size,
acquiring their
higher
0. 42
density
of species of close resemblance
growth period with
aegypti
variability that has not been investigated yet in our
environmental
summarize and combine
followed the
body
group
metabolic sense,
larval
various
three Anopheles the sizespecific
all
content of protein
the
of
depth and larval
extremes, we now can
relation
comparison
manipulated
water
and
Briegel 1994).
or detritus with all the microbial growth on it to feed

upon ( Fish and Carpenter 1982). This even holds for
Culex pipiens( unpubl. research) which can be found in
deep, mostly man- made water containers.

In our experiments food was spread onto the water
with their
surface, and part of it always sunk. Therefore, it was
relatively abundant reserves and usually their single

blood meals per gonotrophic cycle ( Briegel 1990a,
interesting to find thatAnopheles larvae usually did not

survive in depths of more than 2 cm. Obviously, they
This is in clear contrast to the culicine females
unpubl. research).
Within this latter group, Van Handel
were
incapable
of
feeding exclusively
from the
surface
and
ingesting food
consequently depended on
that
explained
few
larvae are
their
by
confined
to shoal biotopes
ability to dive
centimeters,
and
within
can
be
Aly, C. 1988. Filtration rates of mosquito larvae in
suspensions of latex microspheres and yeast cells.
only
dilemma.
energetic
an
suggesting
feed
Outside of this range, breathing at the surface and diving

for food may become lethal for the larvae, probably due
depth for
to exhaustion. However, we could extend the
An.
survival of
An.
albimanus and
gambiae
to
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anophelinae
185
DECEMBER, 1993
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quantities of human blood ingested in Ae. aegypti
L.) (Diptera: Culicidae). Ann. Ent. Soc. Am. 49:
435- 441.
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BSOVE 18( 2): 75- 187 ( 1993)

ISSN 0146- 6429

Volume 18, Number 2

Marc J. Klowden, Editor

North Carolina State

Liverpool School of Tropical Med.

Michigan State University

Med. Vet. Entomol. Lab

Oklahoma State University

New Orleans MCD

San Francisco State University

CH- 8057, Zurich, Switzerland

County Vector Control

Published by the Society for Vector Ecology

facets of the field of vector ecology and related disciplines.

Membership Plus Bulletin$ 45.00

PUBLICATIONS AND BUSINESS OFFICE

Society for Vector Ecology

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SOCIETY FOR VECTOR ECOLOGY

Guidelines for Contributors

In Memoriam- Tuti R. Hadi( 1943 - 1993)

Success in Trinidad, West Indies

Mammals in San Diego

The Oriental Latrine

Public Health Importance

Fabricius 1794) ( Diptera: Calliphoridae),

Mosquito Utilization of Resting Sites at an Urban Residence in Southern California

BULLETIN OF THE SOCIETY FOR VECTOR ECOLOGY

Guidelines for Contributors

scientists and who recommend their

Scientific contributions should be sent to Dr. Marc J. Klowden, Editor, Division of

diskette formatted in MS- DOS is

Microsoft Word, Word Perfect, or

and address of the

Thomas Desmond Mulhern

He was a driving force within the Eastern

If ever the letter" M," as in mosquito, were applicable

careers, one which

Association of Mosquito Control Workers, predecessor

to the American Mosquito Control Association, which

mosquitoes and mosquito

originated in New Jersey. Tommy served the American

Mulhern had two

originated, and the other

Mosquito Control Association as its President in 1968

and 1969 and as its Executive Director during the later

years of his career.

In 1949, Tommy was invited to California to

New Jersey Agricultural Experiment

While in New Jersey,

consider a position with the state' s newly organized

mid-July at the peak of daily temperatures and mosquito

utilizing the New Jersey light

production and was escorted throughout the state to

services within the

benefited the local

dismay him as he shortly returned with wife Helen and

astonishment over the comparative size of

he was an enthusiastic participant in various aspects of

the state, national, and world vector control programs.