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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.


Communications relating to editorial matters and manuscripts
should be addressed to the Editor. Communications relating to galley proofs,
reprints, subscriptions, SOVE membership, change of address, and other matters
should be addressed to the Business Office.

Membership Plus Bulletin$ 45.00


Student Membership$ 22. 50
Institutional Subscription$ 45. 00

Sustaining

Members$ 150.00

PUBLICATIONS AND BUSINESS OFFICE

Society for Vector Ecology


P. O. Box 87

Santa Ana, California 92702 USA

SOCIETY FOR VECTOR ECOLOGY

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

Santa Ana, California 92702 USA

Membership Plus Bulletin$ 45.00


Student Membership$ 2250
Institutional Subscription$ 45.00

Sustaining Members$ 150. 00

BULLETIN

OF THE

SOCIETY FOR VECTOR ECOLOGY


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 non- entomological 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

BULL. SOC. VECTOR ECOL., 18( 2): 77- 84

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

BULL. SOC. VECTOR ECOL.

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

BULL. SOC. VECTOR ECOL.

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

BULL. SOC. VECTOR ECOL.

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

BULL. SOC. VECTOR ECOL.

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

BULL. SOC. VECTOR ECOL.

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

BULL. SOC. VECTOR ECOL.

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

BULL. SOC. VECTOR ECOL.

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.

BULL. SOC. VECTOR ECOL., 18( 2): 85- 98

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

BULL. SOC. VECTOR ECOL.

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

non- random

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

BULL. SOC. VECTOR ECOL.

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 density- dependent 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

BULL. SOC. VECTOR ECOL.

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 density- dependent


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

mosquito- borne 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 density- dependent 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 density- dependent

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.

BULL. SOC. VECTOR ECOL.

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 density- dependent

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

BULL. SOC. VECTOR ECOL.

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 density- dependent

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

BULL. SOC. VECTOR ECOL.

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 non- random

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

BULL. SOC. VECTOR ECOL.

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 density- dependent.
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 density- dependent 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

BULL. SOC. VECTOR ECOL.

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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BULL. SOC. VECTOR ECOL., 18( 2): 99- 104

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.

MATERIALS AND METHODS

al 1991). Statewide surveys of adult ticks on white- tailed


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

BULL. SOC. VECTOR ECOL.

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

BULL. SOC. VECTOR ECOL.

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

non- significant(

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

BULL. SOC. VECTOR ECOL.

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 white- tailed 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 white- tailed 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

BULL. SOC. VECTOR ECOL.

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.
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1992.

Distribution,

and

Sweeney.

Lyme disease

infection in Ixodes dammini ( Acari:

spirochete

Ixodidae)

density,

K.

and

on

white- tailed

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

white- tailed

deer.

J.

Med. Ent. 29: 980- 984.

Ent.( in press).
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and

A. G. Barbour. 1983. Spirochetes in Ixodes dammini


and

mammals

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Hyg.

from Connecticut.

Am. J.

Trop.

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J. L. Benach. 1984. Prevalence

of

the Lyme disease spirochete in populations of whitetailed deer and white- footed mice.

Yale J. Biol.

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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.

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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.

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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,

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49 pp.

Wiesfeld, A. W. Hightower,

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Sharon, M. D., W. A.

DECEMBER, 1993

Blacksburg, 44 pp.
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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

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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.

BULL. SOC. VECTOR ECOL., 18( 2): 105- 108

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

MATERIALS AND METHODS

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

BULL. SOC. VECTOR ECOL.

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

BULL. SOC. VECTOR ECOL.

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.

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size

and more likely to be parous, indicating that they


obtained

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suggested

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blood

Nasci ( 1986)

meals.

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short

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that

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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.

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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-

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BULL. SOC. VECTOR ECOL., 18( 2): 109- 113

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.

MATERIALS AND METHODS


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

BULL. SOC. VECTOR ECOL.

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.

BULL. SOC. VECTOR ECOL.

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

Populations Blood Fed

Populations Non- Blood Fed

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)

Populations Blood Fed

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)

Populations Blood Fed

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

Populations Non- Blood Fed

Populations Non- Blood Fed

112

not

BULL. SOC. VECTOR ECOL.

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.


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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

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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.

BULL. SOC. VECTOR ECOL., 18( 2): 114- 124

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
MATERIALS AND METHODS

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

BULL. SOC. VECTOR ECOL.

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

BULL. SOC. VECTOR ECOL.

6)

sninosnw

DECEMBER, 1993

snyy

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O

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ci,

1) vuviui8.nn siydiapij

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z

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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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N
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08)

iiiagwv8 snivinoiurnu

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601) sninalaivrfsnoiwaia

snos(

snosLCwo1ad

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9)

su8su

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en

en

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a
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i

6S0`i) sad pnu i( ayoaaq sniiydauaads

K =

ra

t7

r
N

v
O

C
8V

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tZ
wi

_
i

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o
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O
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Vg g

az

C7

Marchette

1962).

et al.

Peromyscus

During

M.

of

specimen

single

117

BULL. SOC. VECTOR ECOL.

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

BULL. SOC. VECTOR ECOL.

DECEMBER, 1993

zzz
8)

sun

runf s uvd

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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
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000

8
ar

1(

6 SO` I)

sadpnu!(

ayaaagsnyydouuads

zU

y
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en
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ea
4.

0
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U
V

nz' ^ 3^'
zs

CI

00

8v/

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6

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cI '
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4"

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gg

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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

BULL. SOC. VECTOR ECOL.

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

BULL. SOC. VECTOR ECOL.

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

BULL. SOC. VECTOR ECOL.

DECEMBER, 1993

i)

snalwofrlva a spo

Cyy
8

N
6)

snlnasnw

snyy

E
o

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

BULL. SOC. VECTOR ECOL.

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
in

southern

California. Am. J. Trop. Med. Hyg. 19:

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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County,

California. J. Infect. Dis. 72: 68-

Pasteurella pestis in fleas. V. The experimental

plague vector efficiency of wild rodent fleas

76.

compared with Xenopsylla cheopis, together with


Haddow, J. R., R. Traub,
Distribution of

and

1983.

M. Rothschild.
fleas

ceratophyllid

and notes on

observations on the influence of temperature. Am.

J. Trop. Med. Hyg. 5: 1058- 1070.

their hosts. Material in the collection, with additional

Pp.

notes and maps of the genera.

Rothschild

tophyllidae:

42- 163 in The

Lang, J. D. and W. Wills. 1991. Ecology of sylvatic

Cera-

plague in the San Jacinto Mountains of southern

fleas. The

collection of

family

to the genera and host

key

relation-

California. Bull. Soc. Vector Ecol. 16: 183- 199.

ships, with notes on their evolution, zoogeography

importance,.( R. Traub, M. Rothschild,

Lewis, R. E. 1972. Notes on the geographical distribution

J. F. Haddowm, eds.). Univ. Press, Cambridge,


Academic Press, Inc.( London).

and host preferences in the Order Siphonaptera.


Part 1. Pulicidae. J. Med. Entomol. 9: 511- 520.

D. S. Longanecker.

Lewis, R.E. 1974. Notes on the geographical distribution


and host preferences in the Order Siphonaptera.

andmedical

and

U. K. Distrib.

by

228 pp.
Holdenried, R., F. C. Evans,

1951.

and

Host- parasite- disease

mammalian

in

relationships

community in the

central coast range

Part 3. Hystrichopsyllidae. J. Med. Entomol. 11:


147- 167.

of California. Ecol. Monogr. 21: 2- 13.


Holland, G. P. 1985. The fleas

of

Canada, Alaska
Memoirs

Greenland ( Siphonaptera).
Entomological

Society

of

Canada -

and

of

the

Lewis, R. E., J. H. Lewis, and C. Maser. 1988. The fleas


of the Pacific Northwest. Oregon State Univ. Press,
Corvallis, 296 pp.

No. 130.

Linsdale, J. M. 1946. The California ground squirrel,

Entomol. Soc. Can., Ottawa, Ontario, 631 pp.

a record of observations made on the Hastings


Hopla, C. E. 1980. A study of the host associations
and zoogeography of Pulex.
Pp. 185- 207 in

Natural History Reservation. Univ. Calif. Press,


Berkeley, 475 pp.

Proceed. Intern. Confer. on Fleas ( Traub, R. and


H. E. Stark., eds.), 21- 25 June 1977. A. A. Balkema,
Rotterdam, 300 pp.
Hubbard, C. A. 1947. Fleas of western North America.

1949. Fleas

of

Johnson. 1962. Studies on infectious diseases in


wild animals in Utah. IV. A wild rodent
Peromyscus) plague focus in Utah. Zoo. Res. 1:
341- 361.

Iowa State College Press, Ames, 533 pp.


Hubbard, C. A.

Marchette, N. J., J. B. Bushman, D. P. Parker, and E. E.

the State of Nevada.

Meyer, K. F., R. Holdenried, A. L. Burroughs, and E.


Jawetz.

Bull. S. Calif. Acad. Sci. 48: 115- 128.

1943. Sylvatic

plague

studies.

IV.

Inapparent latent sylvatic plague in ground squirrels


Irons, J. V., S. W. Bohls, D. C. Thurman, Jr.,

1944.

T. Mc Gregor.

Am. J.

Jameson, E. W., Jr.,

and

in central California. J. Infect. Dis. 73: 144- 157.

Probable role of the cat

flea, Ctenocephalides felis, in


murine typhus.

and

Trop.

transmission

of

Med. 24: 359- 362.

Meyer, K. F. and R. Holdenried. 1949. Rodents and


fleas in a plague epizootic in a rural area of

California. Puerto Rico J. Publ. Hlth. Trop. Med.

J. M. Brennan.

1957.

An

24: 201- 209.

environmental analysis of some ectoparsites of


small

forest

mammals

in the Sierra Nevada,

Jameson, E. W., Jr.,


mammals.

and

Miles, V. I., A. R. Kinney, and H. E. Stark. 1957. Fleahost relationships of associated Rattus and native

California. Ecol. Monogr. 27: 45- 54.


H. J. Peeters. 1988. California

Calif. Nat. Hist. Guide No. 52.

Univ.

wild rodents in the San Francisco Bay area of


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.

and

F. M. Prince.

1956.

Studies

on

Vector Contr., 76 pp.

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Nelson, B. C. 1980. Plague


roles

of

various

studies

species

in California- the

Schwan, T. G., D. Thompson, and B. C. Nelson. 1985.

in

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
pp.
Prince, F. M. 1943. Species

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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

studies of reservoirs and

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.

J. Med. Entomol. 20: 558- 564.

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

BULL. SOC. VECTOR ECOL., 18( 2): 125- 132

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


MATERIALS AND METHODS

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

BULL. SOC. VECTOR ECOL.

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.

RESULTS AND DISCUSSION

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 post- flooding) 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 post- flooding, 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

BULL. SOC. VECTOR ECOL.

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

BULL. SOC. VECTOR ECOL.

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

BULL. SOC. VECTOR ECOL.

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

Days Post- Treatment

Figure 4.

Efficacy of various rates of Vectobac 12AS against Culex larvae at Riverside ponds( November
1991).

130

BULL. SOC. VECTOR ECOL.

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

BULL. SOC. VECTOR ECOL.

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

Days Post- Treatment

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

BULL. SOC. VECTOR ECOL.

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

BULL. SOC. VECTOR ECOL., 18( 2): 133- 146

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

BULL. SOC. VECTOR ECOL.

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

BULL. SOC. VECTOR ECOL.

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

BULL. SOC. VECTOR ECOL.

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

BULL. SOC. VECTOR ECOL.

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

BULL. SOC. VECTOR ECOL.

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

BULL. SOC. VECTOR ECOL.

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

BULL. SOC. VECTOR ECOL.

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

BULL. SOC. VECTOR ECOL.

Nicholas J. Richter

and

J. Richard Gorham(

David Faulkner ( SDMNH)

and

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Sychevskaya, V. I. 1981. Synanthropic flies( Diptera)


of the

Lower

reaches of the

Amu- Darya. Entomol.

Rev. 51: 320- 331.


Thomas, H. T.
genera

1951.

1957a.

The life-history and

bionomics of Chrysomyia megacephala ( Fab.).


Ceylon J. Sci.( B) 25: 169- 185.

Some species of the blowfly

Chrysomyia

Hemipyrellia Tnsd.,
southeastern

Wijesundara, D. P.

R- D.,

and

Lucilia

R- D.,

Calliphora R- D. from

Szechuan, China. Proc. Zool. Soc.

Wijesundara, D. P.

1957b. On the longevity of the

adults of Chrysomyia megacephala ( Fab.) under

controlled humidity. Ceylon J. Sci.( B) 25: 187.

London 121: 147.

Wilton, D. P. 1961. Refuse containers as a source of


Tullis, K. and M. L. Goff. 1987. Arthropod
in exposed

carrion

in

succession

a tropical rainforest on

O' ahu

flies in Honolulu and nearby communities. Proc.


Haw. Entomol. Soc. 17: 477- 481.

Island, Hawai' i. J. Med. Entomol. 24: 332- 339.

Zumpt, F. 1965. Myiasis in man and animals in the Old


Tumrasvin, W., H. Kurahashi,

and

R. Kano.

1979.

World. Butterworths, London. 276 pp.

BULL. SOC. VECTOR ECOL., 18( 2): 147- 151

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

MATERIALS AND METHODS

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.

BULL. SOC. VECTOR ECOL.

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

RESULTS AND DISCUSSION

Mosquito Species
for the collection

of

The results regarding the mosquito species occurring

DECEMBER, 1993

in the

July

BULL. SOC. VECTOR ECOL.

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

BULL. SOC. VECTOR ECOL.

TABLE 4.

DECEMBER, 1993

Cumulative list of night- time 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

BULL. SOC. VECTOR ECOL.

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 blood- fed 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

blood- fed

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

BULL. SOC. VECTOR ECOL.

DECEMBER, 1993

species in the collections, five biotic variables were

MATERIALS AND METHODS

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

BULL. SOC. VECTOR ECOL.

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

BULL. SOC. VECTOR ECOL.

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

BULL. SOC. VECTOR ECOL.

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

BULL. SOC. VECTOR ECOL.

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

BULL. SOC. VECTOR ECOL.

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.

BULL. SOC. VECTOR ECOL., 18( 2): 160- 163

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

MATERIALS AND METHODS

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

BULL. SOC. VECTOR ECOL.

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

BULL. SOC. VECTOR ECOL.

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

BULL. SOC. VECTOR ECOL.

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 non- target 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.

BULL. SOC. VECTOR ECOL., 18( 2): 164- 173

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.

BULL. SOC. VECTOR ECOL.

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

BULL. SOC. VECTOR ECOL.

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

BULL. SOC. VECTOR ECOL.

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

BULL. SOC. VECTOR ECOL.

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

BULL. SOC. VECTOR ECOL.

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

BULL. SOC. VECTOR ECOL.

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

BULL. SOC. VECTOR ECOL.

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

BULL. SOC. VECTOR ECOL.

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.

BULL. SOC. VECTOR ECOL.

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.

BULL. SOC. VECTOR ECOL., 18( 2): 174- 187

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 " size- specific 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 size- specific
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

BULL. SOC. VECTOR ECOL.

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

MATERIALS AND METHODS

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
density- dependent
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

BULL. SOC. VECTOR ECOL.

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

size- specific 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

BULL. SOC. VECTOR ECOL.

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

BULL. SOC. VECTOR ECOL.

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

BULL. SOC. VECTOR ECOL.

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

BULL. SOC. VECTOR ECOL.

by - 56

contents

percent, and -

83

percent

for lipid. In

size- specific protein and size- specific 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 " size- specific caloric
content" of protein or

only a minor caloric segment without any correlation.

A different situation was revealed for Ae. aegypti.


Their size- specific 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 (

size- specific 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

BULL. SOC. VECTOR ECOL.

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 size- specific 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

BULL. SOC. VECTOR ECOL.

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 size- specific 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

BULL. SOC. VECTOR ECOL.

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 size- specific 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

BULL. SOC. VECTOR ECOL.

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" size- specific 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

size- specific

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

size- specific

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 size- specific

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

REFERENCES CITED

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anophelinae

185

BULL. SOC. VECTOR ECOL.

DECEMBER, 1993

by

cm

dissolved form. Although this


administering the food in

Ent. Exp. Appl. 6: 55- 62.


Barbosa, P., T. M. Peters, and N. C. Greenough. 1972.
Overcrowding of mosquito populations: responses

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improved the developmental success, the protein and


lipid contents

of the teneral

females

Presumably,

the

of

amount

diluted because it

ignoring
our

the increased

volume of water.

size variation in the pitcher plant mosquito

Wyeomyia smithii( Diptera: Culicidae). Ann. Ent.

surface

certain anophelines.

In Ae.

aegypti

larger

with much

there

are similar principles although

They

margins.

could

dive, feed,

and

successfully develop down to at least 14 cm. With the


food dissolved,
increased

at

advantage

to the

feeding

to the

domain.

The efficiency of food


shallow

develop
due to

reduced

acquisition might

Anopheles

pools where"

when

the

secondary

level is dras-

water

evaporation.

be crucial,

Dahl

et al. (

1988)

gave a detailed anatomical description of the larval


feeding mechanisms, while Aly(
Mulla( 1989) determined

1988)

were unaffected

Anopheles

species appear

by

much

biosynthetic
anophelini,

J. Med. Ent. 30: 975- 985.

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Acknowledgments

mosquitoes (

appreciate

the reliable

assistance given

by Mrs. R. Haigis with some biochemical analyses and


the

generous

support of

the Swiss National Science

Foundation to H. Briegel. The


comments

by

the

acknowledged.

Linnaeus) ( Diptera:

of culicini and

respectively.

We greatly

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and

acquisition

capabilities of

Culicidae).

and yet

might furnish a plausible explanation for the different


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to have

Briegel, H. and E. Horler. 1993. Multiple blood meals

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Rashed

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and

species- specific

The latter

rates

Metabolic relationship between

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every

can

1990a.

female body size, reserves, and fecundity of Ae.


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Briegel, H.

depth, obviously rendering


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density- tolerance, revealed a partial


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thus
deepening its larval
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crowding"

Soc. Am. 85: 274- 281.

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1992.

Nevertheless,

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C. M. Holzapfel.

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W. E.

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