Susceptibility of Fleas
Dryden, DVM, PhD
to Control Agents:
Development of
a Monitoring System
T
he development and marketing of highly effective, residual
insecticides/insect growth regulators during the past few
years has dramatically changed the way pet owners treat
their pets for fleas. The increased reliance on single-compound, on-
animal, or oral application systems to control fleas has been accom-
panied by a corresponding decrease in the treatment of premises and
an indifferent attitude toward flea biology and epidemiology.
Although these compounds are remarkably effective, there is a con-
cern that an overreliance on any single compound to control insects
may lead to the development of insecticide resistance.
Resistance to insecticides was defined by the World Health
Organization (WHO) as the development of an ability in a strain of
some organism to tolerate doses of a toxicant that would prove
lethal to a majority of individuals in a normal population of the
same species.1 Resistance has a genetic basis and is the result of a
change in the genetic composition of a population as a direct result
of the selective effects of a toxicant.
History of Insecticide Resistance
In 1951 Pulex irritans was suggested as becoming resistant to
DDT after sprays were applied to control malarial mosquitoes.2
Soon thereafter, many locations, mainly in the tropics, reported
control failures. Laboratory bioassays confirmed that insecticide
selection pressure had forced genetic evolution of flea populations
with resistance as the outcome.2 By 1983, eight species of fleas,
including the fleas of public health importance Ctenocephalides felis,
P. irritans, and Xenopsylla cheopis, had demonstrated resistance to
many of the insecticides used in flea control.3 In addition, labora-
tory selection studies have produced resistance ratios (the ratio of
lethal doses of a resistant strain to lethal doses of a susceptible
strain) of 12 and 108 over unselected controls against malathion in
two separate experiments.4,5 A field-collected strain of cat fleas
from Florida had resistance ratios of 26 and 28 over a laboratory
strain from California for malathion and bendiocarb, respectively.6,7
Cat fleas are thought to be resistant to more chemical families than
any other flea.8
TNAVC, January 2000
Michael W.
Associate Professor of Veterinary Parasitology
Department of Diagnostic Medicine/Pathobiology
College of Veterinary Medicine
Kansas State University
Manhattan, KS, USA
Alberto B. Broce, PhD
Professor of Entomology
Department of Entomology
Kansas State University
Manhattan, KS, USA
Determining the Cause of Product Failures
Because it is understood that development of resistance in cat
flea populations is always a possibility, veterinarians should not be
quick to blame resistance for all perceived control failures. Veteri-
narians must always be aware that several factors other than resist-
ance may be involved when treatment is perceived to have failed.
In clinical practice it may be difficult or impossible to deter-
mine if a particular product failed because of resistance. Perceived
failure in practice situations could be from lack of owner compliance,
environmental burden of the pest, presence of untreated hosts,
bathing of the treated pet, vomiting of oral medications, or actual
resistance. It can be difficult to determine the reason for perceived
product failure without considerable investigation. Even in the labo-
ratory a number of factors can affect the results of a laboratory bioas-
say, such as rearing conditions, immobilization method, age of organ-
ism, shipping stresses, treated substrate, temperature, or humidity.
Terminology for Describing Product Failures
Another problem practitioners encounter is a misunderstanding
of terms used to describe failures. One such term is reduced suscep-
tibility. The reasons for reduced susceptibility may vary depending
upon whether one is referring to a laboratory bioassay, an on-animal
efficacy trial, or a field event. This term simply refers to a popula-
tion of organisms that has a higher survival rate than expected when
exposed to a particular compound, regardless of the cause.
Tolerance refers to the response by a population of organisms to
a toxicant relative to other species. It is the innate genetic predispo-
sition of a population. Examples of tolerance include ticks being
more tolerant of imidacloprid than fleas are, and Trichuris vulpis being
more tolerant of pyrantel pamoate than is Ancylostoma caninum.
These are conditions that exist irrespective of selection pressure.
Insecticide resistance is the ability of an insect population to sur-
vive doses and concentrations of an insecticide that previously were
lethal to that population. Insecticide resistance is a genetic change in
a population caused by the elimination of the susceptible individuals,
leaving only the resistant individuals to breed. Resistance is a form of
Suppl Compend Contin Educ Pract Vet Vol. 22, No. 4(A), 2000
time-compressed evolution. Classic evolution is a gradual selection of
more competitive morphologic, physiologic, or behavioral traits,
whereas resistance selection is a rapid evolutionary process. The appli-
cation of the insecticide results in an instantaneous widespread eco-
logic disaster on the population. Individuals with morphologic, physi-
ologic, or behavioral traits that allow survival remain to reproduce.
Although a great deal of knowledge has been gained concerning
insecticide resistance in other insect species, unfortunately in fleas
the extent, prevalence, or clinical significance of insecticide resist-
ance to any particular insecticide has not been determined. Although
it is apparent that insecticide resistance has been detected in various
cat flea populations, there currently is no active program to monitor for
resistance. In addition, no standard bioassay system has been used to
compare strains, and there are no data to indicate what level of sur-
vivability in a bioassay corresponds to control failure on animals.
The Flea Susceptibility Monitoring Team
In September 1999 a Flea Susceptibility Monitoring Team was
organized by Bayer, Animal Health. The team includes Byron Blagburn
(Auburn University, USA), Michael Dryden (Kansas State University,
USA), Dennis Jacobs (Royal Veterinary College, London, UK),
Michael Rust (University of California at Riverside, USA), Ian Den-
holm and Martin Williamson (Institute of Arable Crops Research
[IACR], Rothamsted, UK), Heinz Mehlhorn (Heinrich Heine Univer-
sity, Düsseldorf, Germany), and the Research and Development Divi-
sion of Bayer Animal Health (United States, Germany, Australia).
The team was formed to establish protocols and procedures to
monitor cat flea susceptibility to imidacloprid, determine clinical
significance of reduced susceptibility that may be found in the
future, and formulate plans to steward imidacloprid into the future.
Monitoring for Insecticide Resistance
Monitoring for insecticide resistance in wild populations of pests
has often been used to describe a continuous inspection of levels of
efficacy of an insecticide applied to such populations. The first step
of a monitoring program should be to obtain baseline data before
resistance appears and the use of the insecticide is widespread. How-
ever, if baseline data cannot be obtained a priori, then data from sev-
eral laboratory strains should be used to establish such a baseline.
Obviously, these should be insecticide-susceptible strains that have
not been exposed to insecticides during their laboratory rearing. One
would hope that the dose-responses of these lab strains would cover
the range of original variability found among wild populations.
The first step proposed in establishing baseline data is to estab-
lish a practical, repeatable, and reliable bioassay system to monitor
for imidacloprid susceptibility in five separate laboratories. The
monitoring team will do this by determining the LC50 (median
lethal concentration) and LC95 of five laboratory strains of cat fleas
using a larval development inhibition test. In addition, the team
will establish the range of LC50 and LC95 of the same five strains
using an adult in vitro bioassay. Data from all laboratories will then
Suppl Compend Contin Educ Pract Vet Vol. 22, No. 4(A), 2000
be combined to establish LC50 and LC95 to be used in a discriminat-
ing dose bioassay. A discriminating dose bioassay will then be used
for large-scale screening of field populations of cat fleas. A discrim-
inating dose bioassay will allow for more rapid screening of larger
numbers of cat flea strains. It is planned that during the first year the
five laboratories will randomly screen up to 150 flea isolates (indi-
vidual collections of fleas from infested dogs or cats in veterinary
practices) in the United States and 100 in Europe.
During this screening process a population will be considered
potentially resistant if the mortality at the discriminating dosage falls
outside the confidence limits of the normal range for the five baseline
strains. The numbers of any such field strain determined to be poten-
tially resistant will be increased in the laboratory, and their dose-
response will be determined in a classic bioassay. A population will be
considered resistant if its LD50 (median lethal dose) and/or LD95 is sig-
nificantly greater (outside the 95% confidence interval) than the
LD50 and/or LD95 of the standard value for the combined strains.
Most programs for monitoring insecticide resistance in other
species of insects do not attempt to correlate the dose-response data
to field performance (i.e., efficacy) of the insecticide tested. This is
an important question when monitoring for the development of
insecticide resistance, mainly for two reasons: first, there is no infor-
mation on how well the dose-response data equate to the on-host
efficacy of the chemical in question; second, the validity of using
dose-response of strains reared under laboratory conditions for a
long time is questionable. Therefore, once a strain has been deter-
mined to be resistant in the classic bioassay, the team will conduct
on-host efficacy evaluations and simulated home environment trials
of the resistant isolate(s) using standard doses of imidacloprid to
determine if resistance levels found in laboratory bioassays have any
clinical significance.
Finally, because virtually nothing is known concerning the
resistance mechanisms of resistance in fleas, the team will attempt
to determine biochemical and genetic mechanism(s) of resistance.
References
1. Ferrari JA: Insecticide resistance, in Beaty BJ, Marguardt WC: The Biolo-
gy of Disease Vectors. Niwot, Colo, University Press of Colorado, 1996, pp
512–529.
2. Brown AWA, Pal R: Insecticide Resistance in Arthropods, ed 2. Geneva,
WHO, 1971, p 491.
3. Georghiou GP, Mellon RP: Pesticide resistance in time and space, in
Georghiou GP, Saito T (eds): Pest Resistance to Pesticides. New York,
Plenum Press, 1983, pp 1–46.
4. Collart MG, Fink WF: Development of resistance to malathion in cat flea
(Siphonaptera: Pulicidae). J Econ Entomol 79:1570–1572, 1986.
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7. El-Gazzar LM, Patterson RS, Koehler PG: Comparisons of cat flea
(Siphonaptera: Pulicidae) adult and larval insecticide susceptibility. Fla
Entomol 71:359–363, 1988.
8. Rust MK: Insecticide resistance in fleas, in Knapp FW (ed): Intl Symp
Ectoparasites of Pets. Lexington, KY, University of Kentucky, 1993, pp 18–26.
International Flea Control Symposium