Accumulation and persistence of flea larvicidal activity in the immediate environment of cats treated with imidacloprid

To investigate the persistence of flea larvicidal activity in the immediate environment of cats treated with imidacloprid, eggs of the cat flea Ctenocephalides felis felis Bouché (Siphonaptera: Pulicidae), from untreated donor cats, were incubated on samples of fleece blanket taken from the floor of cages used by treated or untreated cats for a total of 10 or 20 6-h periods over 2-4 weeks, respectively. Sufficient imidacloprid accumulated during these periods to reduce the emergence of adult fleas by 94.7-97.6% when the blankets were tested after 18 weeks' storage at room temperature. A typical laundry procedure (washing with detergent at 50 degrees C and low temperature tumble drying) removed this biological activity. Unwashed control blankets did not support the flea life-cycle as effectively as washed blankets or a sand substrate.     

Comparative efficacy on dogs of a single topical treatment with fipronil/(S)-methoprene or weekly physiological hygiene shampoos against Ctenocephalides felis in a simulated flea-infested environment

Flea infestations of pets continue to persist due to the lack of knowledge of flea biology and ecology. It is not unusual that pet owners believe regular hygiene, such as shampooing their dogs can replace regular insecticidal treatment. The objective of this study was to compare in a flea simulated environment, modelling exposure similar to that found in a home, that the use of regular physiological shampoo does not control fleas adequately when compared to a long acting topical formulation. Three groups of six dogs were formed: one untreated control group, one group treated monthly with the topical formulation of fipronil/(S)-methoprene, and a third group treated weekly with a hygiene shampoo. All dogs were infested with adult unfed Ctenocephalides felis fleas (200±5) on Days -28 and -21. Each animal's sleeping box was fitted with a plastic cup mounted to the inside roof of the box. The sleeping bench of each animal was covered with a carpet to accommodate flea development. The dogs were maintained in their kennels throughout the study. In order to maintain the environmental flea challenge, C. felis pupae (100±5) were placed in the plastic cup in each animal's sleeping box on Days -14, -7, 0, 7, 14, 21, 28, 35 and 42. The dogs were combed and fleas counted weekly on Days -1, 3, 10, 17, 24, 31, 38, 45, and 51. The fleas were placed immediately back on the dogs. On Day 60, fleas were counted and removed. Flea infestations in the untreated control group at each count averaged between 46.2 and 74.2 fleas throughout the study. The average number of fleas infesting dogs was significantly different (p<0.05) between the untreated and the two treatment groups and between the two treatment groups at all counts throughout the two months study (Days 3, 10, 17, 24, 31, 38, 45, 51 and 60). The efficacy was never below 99.1% in the fipronil/(S)-methoprene group, and efficacy in the shampoo group was never above 79.2%. Weekly shampooing in treatment group 3 was intentionally delayed after Day 42, to evaluate wether missing a weekly bath would affect the flea population. The Day 48 data indicate that forgetting or delaying a single weekly shampooing resulted in a clear increase in flea numbers and a significant decrease in efficacy from 68.2% to 34.8%. The fipronil(S)/methoprene treatment allowed a continuous control as demonstrated by the high efficacy against fleas, and also the number of flea-free dogs on seven of the nine weekly assessments, in spite of what was essentially a continuous flea challenge model.     

Large-scale monitoring of imidacloprid susceptibility in the cat flea, Ctenocephalides felis

Although on-animal topical treatment with compounds such as imidacloprid has revolutionized the control of the cat flea, Ctenocephalides felis (Bouché) (Siphonaptera: Pulicidae), the development of insecticide resistance is a continuing threat. As part of a highly co-ordinated and unprecedented resistance monitoring programme for C. felis, 1437 flea isolates were collected by veterinary clinics in Australia, Germany, France, the U.K. and 29 states in the U.S.A. from 2002 to 2009. About 65% of the collections were made from June to October each year and 71% of the collections were from cats. Collections of flea eggs were sent to one of five different laboratories, where they were tested with a diagnostic dose of imidacloprid (3 p.p.m.) applied to larval flea-rearing medium. Of the 1437 collections received, 1064 contained adequate numbers of eggs for testing. Of these isolates, untreated eggs failed to hatch in 22.7% and were not considered valid bioassays. Survival rates >5% and development of adult fleas (a threshold for further testing) occurred in only 22 isolates. They were re-tested with the same diagnostic dose and none produced >5% adult emergence. Complete dose-response bioassays were performed on three of the isolates that had triggered a second test and produced slopes, intercepts and LC(50) values similar to those for existing susceptible laboratory strains. Results confirmed sustained susceptibility of C. felis to imidacloprid, despite its widespread use for over a decade.     

Prevalence of flea infestation in dogs and cats in Hungary combined with a survey of owner awareness

A survey was conducted in order to gain current information on flea species (Siphonaptera: Pulicidae) infesting dogs and cats living in urban and rural areas of Hungary, along with data on the factors that affect the presence, distribution and seasonality of infestation. In addition, owner awareness of flea infestation was evaluated. Practitioners in 13 veterinary clinics were asked to examine all dogs and cats attending the clinic and to collect fleas, when present, on 2 days in each month from December 2005 to November 2006. They also completed a questionnaire for each animal examined. A total of 319 dogs (14.1%) were found to be infested; the highest prevalence (27.1%) of infestation on dogs occurred in August and the lowest (5.4%) in May. Prevalence of fleas on cats was higher (22.9%); the highest (35.0%) and lowest (8.1%) prevalences occurred in July and April, respectively. Fleas were more prevalent in rural (387/1924 animals, 20.2%) than in urban (161/1343 animals, 12.0%) areas. Three species, Ctenocephalides felis (Bouché), Ctenocephalides canis (Curtis) and Pulex irritans L., were found. On dogs, the prevalence of C. canis alone was 53.0%, whereas that of C. felis alone was 36.0%. Only 19 specimens of P. irritans were found on 14 dogs from rural habitats only. Prevalence of C. felis only on cats was 94.3%; the remaining cats were infested with either C. canis or with mixed infestations of C. felis and C. canis. More than half (51.4%) of the owners of infested dogs and cats had not used flea control products in the past year or more, and five times as many owners in rural than urban areas had not used flea control products in the same period. Very few owners reported having attempted to kill fleas in their animals' environment; instead, they believed that fleas were acquired from other cats or dogs.     

Establishment of the cat flea (Ctenocephalides felis felis) on the ferret (Mustela putorius furo) and its control with imidacloprid

As the ferret, Mustela putorius furo L. (Carnivora: Mustelidae), is becoming increasingly popular as a pet animal and as it is susceptible to the cat-flea, Ctenocephalides felis felis Bouché (Siphonaptera: Pulicidae), an experimental model was established for evaluating insecticidal treatments on this host. A high establishment rate (76.7-91.8%) was recorded when 60 unfed adult C. felis were placed on ferrets. This provided an adequate infestation for chemotherapeutic evaluation without causing undue discomfort to the host. Twelve ferrets were allocated to two groups matched for sex and individual ability to sustain a flea population. One group was treated topically with an imidacloprid spot-on formulation at a dose rate of 10 mg/kg body-weight on Day 0. All ferrets were infested with C. felis on Days -1, 7, 14, 21 and 28, and flea counts were performed 8 and 24 h post-treatment and one day after each subsequent infestation. Fleas were removed at all but the 8 h count (when they were returned to their host). Flea burdens were reduced by 95.3% (P < 0.001) within 8 h of treatment and 100% efficacy was recorded at 24 h. At 1, 2, 3 and 4 weeks post-treatment, protection against re-infestation was 92.9% (P < 0.001), 55.7% (P < 0.02), 18.3% (NS) and 7.4% (NS), respectively. Thus, at this dose rate, imidacloprid gave excellent efficacy against a resident C. felis population and provided a high level of residual activity for at least one week after treatment.     

Advances in the control of Ctenocephalides felis (cat flea) on cats and dogs

Cat fleas are the most important ectoparasite of cats and dogs worldwide. During the past ten years, topical and oral applications of insecticides such as fipronil, imidacloprid, lufenuron and, most recently, selamectin have revolutionized cat-flea control. Recent studies show that these therapies eliminate the need to treat indoor and outdoor environments, and their use markedly reduces the severity and prevalence of flea allergic dermatitis. Surveys have yet to reveal the development of insecticide resistance to these chemical compounds. Extending the longevity of these effective host-targeted therapies should be a major goal of the veterinary community.     

Notes on fleas (Siphonaptera) in plague foci on the Tay Nguyen plateau (Vietnam)]

The fleas of wild and commensal small mammals, domestic animals (dogs, cats) and free-living flea forms in houses have been collected in plague nidi of Tay Nguyen plate, Dak-Lak province, Vietnam. Pulex irritans, Ctenocephalides felis felis, Ct. felis orientis were found in the houses on dogs, cats and on the ground floor. Commensal rats in populated areas were infested by Xenopsylla cheopis and rarely by Lentistivalius klossi. The agricultural zone was inhabited by both home and wild animals such as commensal, savannah and forest-dwelling small mammals. The flea fauna of this zone is presented by X. cheopis and L. klossi. In the tropical forest surrounding villages four of the flea species were found: X. vexabilis, a specific parasite of the forest-dwelling rat Berylmys berdmorei, L. klossi found on six species of forest small mammals, Acropsylla girshami from Berylmys bowersii and Pariodontis subjugis from Hystrix brachyura. The agricultural zone is the most possible place of commensal and forest-dwelling small mammals contact, where the latter can get plague microbe.     

Systemic efficacy of nodulisporic acid against fleas on dogs

Nodulisporic acid A (NSA) is a novel natural product from a new structural class that was shown previously to have insecticidal activity against blowfly larvae. To determine if there was useful systemic efficacy against fleas (Ctenocephalides felis). NSA was evaluated in an artificial membrane flea feeding device and in dogs. In the artificial membrane flea feeding device, adult C. felis were allowed to feed on bovine blood containing various concentrations of NSA through a Parafilm membrane. NSA killed the fleas with a 50% lethal concentration of 0.68 microg/ml and was approximately 10-fold more potent than the systemic insecticide ivermectin. In the initial probe dog test, a single beagle was challenged with 100 C. felis before oral dosing with 15 mg/kg of NSA. Flea counts conducted at 72 hr postdosing showed an 88% reduction relative to control. Re-challenge of the same dog at 5 days postdosing showed 50% reduction of fleas at day 7, demonstrating some residual flea activity. In a confirmatory study, 8 dogs were challenged with 100 fleas just before oral dosing with 15 mg/kg of NSA (4 dogs) or vehicle (4 dogs). There was 99% reduction of fleas at 48 hr postdosing in the NSA-treated dogs relative to control. Additional challenges with 100 fleas were performed on these 8 dogs at 48-hr intervals to determine the duration of efficacy, and there was 97, 51, and 0% reduction of fleas relative to control on days 4, 6, and 8, respectively. No adverse effects were observed in the dogs in these studies. These data show that NSA has potent oral activity in the dog for the control of fleas, while lacking overt mammalian toxicity.     

Egg production, larval development, and adult longevity of cat fleas (Siphonaptera: Pulicidae) exposed to ultrasound

Adult cat fleas, Ctenocephalides felis felis (Bouché), on cats (Felis catus) were exposed to emissions from an ultrasonic flea collar worn by the cat. No significant differences were found in total numbers of eggs produced per day (mean = 524 control, 614 treatment), in length of larval development time (mean = 7.7 d control, 7.7 d treatment), or in total daily pupal production (mean = 485 control, 445 treatment) between the treatment and the control groups. Tests off the host were conducted to determine whether ultrasonic exposure caused mortality in adult fleas; no significant differences were found in daily mortality between the treated and control fleas during 1 wk of exposure.     

An attempt to protect cats against infestation with Ctenocephalides felis felis using gut membrane antigens as a vaccine

Cats (n = 5) were vaccinated with membrane antigens extracted from the gut of unfed fleas (Ctenocephalides felis felis) together with Quil A and RIBI as adjuvants. Five unvaccinated cats were retained as controls. All the cats were infested on 6 separate occasions with fleas (46–250 per challenge). Protection was assessed from the number of fleas retrieved and the fecundity of the female fleas, measured as the number of developed oocytes contained in the reproductive tract. Cats injected with gut membrane antigens had significantly elevated levels of anti-flea antibodies in their sera, but they were neither protected significantly against infestation with fleas nor was the apparent fecundity of fleas which had fed on vaccinated cats decreased. The possible reason why gut membrane antigens failed to protect cats against fleas are discussed.     

First blood meal of Ctenocephalides canis (Siphonaptera: Pulicidae) on dogs: time to initiation of feeding and duration

Two experiments were conducted on dogs to evaluate interval to initiation and duration of the first blood meal of Ctenocephalides canis (Curtis). Percentage of fed male and female fleas was calculated for fleas held on dogs for 5, 15, 30, 60 min, 6, and 24 hr. Duration of first blood meal was also measured for individual fleas confined on dogs. When fleas were free in the hair coat, 21.2% had begun blood feeding within 5 min. After 1 hr, 72.5% of fleas had fed. After 6 hr, 95.2% of males and 100% of females had taken a blood meal, and 24 hr after deposition all fleas had fed. There was no significant difference between the 2 sexes. The mean delay between deposition and biting for fleas that began feeding within 15 min was 2 min 52 sec +/- 3 min 2 sec for female fleas and 3 min 8 sec +/- 2 min 45 sec for males. The mean duration of female and male meals was 5 min 3 sec +/- 3 min 41 sec and 6 min 9 sec +/- 6 min 8 sec, respectively. There was no significant difference between the 2 sexes. The dog flea took its blood meal on dogs more slowly than the cat flea did on cats; this meal was significantly longer for Ctenocephalides felis felis (Bouche) than for C. canis.     

Results of a European multicentric field efficacy study of fipronil-(S) methoprene combination on flea infestation of dogs and cats during 2009 summer

Despite the increasing number of ectoparasiticides for pets and their use, flea infestations of cats and dogs are still widespread in Europe. It is therefore important to assess the maintenance of efficacy of the ectoparasiticides for cats and dogs. The present studies aimed to evaluate the efficacy of monthly treatments using a fipronil/(S)-methoprene combination spot-on (Frontline Combo) on dogs and cats from private veterinary clinics located in seven European countries. The survey was conducted for three months during the flea season 2009. A total of 233 dogs and 180 cats were included. Each animal was treated at Days 0 (Day 0), 30 (D30) and 60 (D60) at the vet clinics. For each animal, at least three flea counts were performed at D0, D30 and/or D60 and/or day 90 (Day 90) in order to evaluate the prevalence of flea infestation and the efficacy of control. At the beginning of the study the prevalence of infested animals was 41.63% (97/233) in dogs and 47.22% (85/180) in cats. At D90, the number of dogs remaining infested fell to 8/211 therefore 91.75% became flea-free. The number of infested cats fell from 85 to 9/173 at D90 therefore 89.41% were cured. All animals still infested at Day 90 were living under epidemiological conditions that favour heavy flea burdens. These results are similar or better to previous studies, indicating the continuous high level of efficacy for fipronil 10 years after launch.     

Out-of-Africa,human-mediated dispersal of the common cat flea,Ctenocephalides felis: The hitchhiker’s guide to world domination

The cat flea (Ctenocephalides felis) is the most common parasite of domestic cats and dogs worldwide. Due to the morphological ambiguity of C. felis and a lack of — particularly largescale — phylogenetic data, we do not know whether global C. felis populations are morphologically and genetically conserved, or whether human-mediated migration of domestic cats and dogs has resulted in homogenous global populations. To determine the ancestral origin of the species and to understand the level of global pervasion of the cat flea and related taxa, our study aimed to document the distribution and phylogenetic relationships of Ctenocephalides fleas found on cats and dogs worldwide. We investigated the potential drivers behind the establishment of regional cat flea populations using a global collection of fleas from cats and dogs across six continents. We morphologically and molecularly evaluated six out of the 14 known taxa comprising genus Ctenocephalides, including the four original C. felis subspecies (Ctenocephalides felis felis, Ctenocephalides felis strongylus, Ctenocephalides felis orientis and Ctenocephalides felis damarensis), the cosmopolitan species Ctenocephalides canis and the African species Ctenocephalides connatus. We confirm the ubiquity of the cat flea, representing 85% of all fleas collected (4357/5123). Using a multigene approach combining two mitochondrial (cox1 and cox2) and two nuclear (Histone H3 and EF-1α) gene markers, as well as a cox1 survey of 516 fleas across 56 countries, we demonstrate out-of-Africa origins for the genus Ctenocephalides and high levels of genetic diversity within C. felis. We define four bioclimatically limited C. felis clusters (Temperate, Tropical I, Tropical II and African) using maximum entropy modelling. This study defines the global distribution, African origin and phylogenetic relationships of global Ctenocephalides fleas, whilst resolving the taxonomy of the C. felis subspecies and related taxa. We show that humans have inadvertently precipitated the expansion of C. felis throughout the world, promoting diverse population structure and bioclimatic plasticity. By demonstrating the link between the global cat flea communities and their affinity for specific bioclimatic niches, we reveal the drivers behind the establishment and success of the cat flea as a global parasite.     

Horizontal transmission of Rickettsia felis between cat fleas, Ctenocephalides felis

Rickettsia felis is a rickettsial pathogen primarily associated with the cat flea, Ctenocephalides felis. Although laboratory studies have confirmed that R. felis is maintained by transstadial and transovarial transmission in C. felis, distinct mechanisms of horizontal transmission of R. felis among cat fleas are undefined. Based on the inefficient vertical transmission of R. felis by cat fleas and the detection of R. felis in a variety of haematophagous arthropods, we hypothesize that R. felis is horizontally transmitted between cat fleas. Towards testing this hypothesis, flea transmission of R. felis via a bloodmeal was assessed weekly for 4 weeks. Rhodamine B was used to distinguish uninfected recipient and R. felis-infected donor fleas in a rickettsial horizontal transmission bioassay, and quantitative real-time PCR assay was used to measure transmission frequency; immunofluorescence assay also confirmed transmission. Female fleas acquired R. felis infection more readily than male fleas after feeding on a R. felis-infected bloodmeal for 24 h (69.3% and 43.3%, respectively) and both Rickettsia-uninfected recipient male and female fleas became infected with R. felis after cofeeding with R. felis-infected donor fleas (3.3-40.0%). Distinct bioassays were developed to further determine that R. felis was transmitted from R. felis-infected to uninfected fleas during cofeeding and copulation. Vertical transmission of R. felis by infected fleas was not demonstrated in this study. The demonstration of horizontal transmission of R. felis between cat fleas has broad implications for the ecology of R. felis rickettsiosis.     

Palatability and toxicity of fipronil as a systemic insecticide in a bromadiolone rodenticide bait for rat and flea control

Vector control in plague-infested areas requires a simultaneous killing of rodents and their fleas. We investigated the efficacy of a combination of a systemic insecticide, fipronil, in a rodenticide bait formulation under laboratory conditions. Four different concentrations of fipronil (0.05%, 0.005%, 0.0005% with acetone as a solvent, and 0.05% with propylene glycol as a solvent) and two controls (solvents only) were combined with the rodenticide bait (crushed organically grown wheat with 0.005% bromadiolone). Each concentration was offered together with an untreated non-poisonous challenge bait to 10 singly caged Rattus rattus L., each with 100 rat fleas Xenopsylla cheopis Rothschild (Siphonaptera: Pulicidae) in the nest. Treated bait consumption was relatively low and an unsatisfactory rat mortality of around 50% only was obtained in all tests. The palatability of the bait, however, was not affected by the fipronil concentration. Even at the lowest fipronil concentration, average flea mortality was still above 95%, and doses of more than I mg fipronil per kg rat body weight gave a nearly complete kill of fleas. Fipronil can be highly effective as a systemic insecticide to for flea control, provided that a more attractive bait base for roof rats is used.     

Comparative efficacy on dogs of a single topical treatment with the pioneer fipronil/(S)-methoprene and an oral treatment with spinosad against Ctenocephalides felis

In the study reported here, the pioneer fipronil/(S)-methoprene topical product (FRONTLINE^® PLUS, Merial Limited, Duluth, GA) was compared to the oral spinosad product (COMFORTIS^® Elanco, Greenfield, IN) for efficacy against adult fleas and preventing egg production. The product presentations, doses and labelling were the one applicable in the USA. Using a standard protocol, 200 cat fleas of mixed sex were applied to dogs on Days 1, 7, 14, 21, 28, 35, and 42. Dogs were combed to remove fleas 24 hours post-infestation, the fleas were counted, collected, and then reapplied to each dog following completion of their respective count. At 48 hours post-infestation, comb counts were performed and fleas were removed. No fleas were collected from any dog in the fipronil/(S)-methoprene group at any 24 or 48 hours post-infestation assessment throughout the six weeks study, yielding a preventive efficacy of 100%. For the spinosad treatment, efficacy was 100% at 24 hours and 48 hours through Day 16, and thereafter declined. The results observed in the spinosad-treated dogs were highly variable between animals. At the 24 and 48 hours counts following the Day 21 infestation, only five of eight spinosad-treated dogs (62.5%) were flea-free. Following the Day 28 infestation, spinosad efficacy fell to 85% and 89%, for the 24 hours and 48 hours counts, and only two dogs (25%) were flea free, compared to 100% flea-free dogs in the fipronil/(S)-methoprene group. No fleas were collected from the fipronil/(S)- methoprene treated dogs throughout the entire study, therefore, no eggs were collected at any time from any dog in the group. However, in the spinosad group adult fleas were found on dogs starting on Day 21 and by Day 30, 42 eggs were collected from one dog that had 107 adult fleas counted at 48 hours. At Day 37 and Day 49, more than 100 eggs were collected from each dog in the spinosad-treated and control groups.     

氟虫腈、吡虫啉作为黑翅土白蚁诱杀药剂的效果

毒力测定结果表明,0.025~0.4μg/mL氟虫腈和吡虫啉分别在药后3 d和5 d对黑翅土白蚁Odontotermes formosanus表现出明显的毒杀效果,氟虫腈和吡虫啉药后1 d的LC50分别为药后5 d的509倍和63.8倍,2种药剂对黑翅土白蚁的毒杀效果均比较缓慢。毒性传递试验表明,0.5μg/g毒沙处理白蚁1 h后,氟虫腈和吡虫啉的致死毒性均可被传毒白蚁传递给受毒白蚁。驱避作用试验表明,50μg/mL氟虫腈对黑翅土白蚁无明显的驱避作用,而50μg/mL吡虫啉对黑翅土白蚁表现出了明显的驱避作用。可见,2种供试药剂中,氟虫腈是较理想的白蚁诱杀药剂。     

Efficacy of a fipronil bait in reducing the number of fleas (Oropsylla spp.) infesting wild black‐tailed prairie dogs

Bubonic plague (Yersinia pestis) is a deadly zoonosis with black‐tailed prairie dogs (Cynomys ludovicianus) as a reservoir host in the United States. Systemic insecticides are a promising means of controlling the vectors, Oropsylla spp. fleas, infesting these prairie dogs, subsequently disrupting the Y. pestis cycle. The objective of this study was to conduct a field trial evaluating the efficacy of a grain rodent bait containing fipronil (0.005%) against fleas infesting prairie dogs. The study was performed in Larimer County, CO, where bait was applied to a treatment area containing a dense prairie dog population, three times over a three‐week period. Prairie dogs were captured and combed for fleas during four study periods (pre‐, mid‐, 1^st post‐, and 2^nd post‐treatment). Results indicated the use of bait containing fipronil significantly reduced flea burden. The bait containing fipronil was determined to reduce the mean number of fleas per prairie dog >95% for a minimum of 52 days post‐initial treatment application and 31 days post‐final treatment application. These results suggest the potential for this form of treatment to reduce flea population density on prairie dogs, and subsequently plague transmission, among mammalian hosts across the United States and beyond.     

Grooming and control of fleas in cats

Oral grooming is common in cats, as in rodent and bovid species where grooming has been shown to be effective in removing lice and ticks. In Experiment 1, we examined the effectiveness of oral grooming in removing fleas which are the main ectoparasite of cats. Elizabethan collars (E-collars) which prevented grooming were fitted on nine cats in a flea-infested household and 3 weeks later, flea numbers on these cats were compared with nine control cats in the same household. Flea numbers dropped in the control cats reflecting an apparent drop in adult fleas in the environment, but in the E-collar cats, flea numbers did not drop, and were about twice as numerous as in control cats. The significantly greater number of fleas on the E-collar cats was attributed to their inability to groom off fleas. In Experiment 2, videotaping of nine different cats from the flea-infested household revealed that these cats groomed at about twice the rate of 10 similarly videotaped control cats from a flea-free colony. These results reveal that flea exposure can increase grooming rate in cats and that grooming is effective in removing fleas.     

Effect of temperature and the synergist piperonyl butoxide on imidacloprid toxicity to the cat flea (Siphonaptera: Pulicidae)

The toxicity of imidacloprid to cat fleas on glass was investigated at 20, 26, 30, and 35 degrees C. Imidacloprid was most toxic to adult cat fleas at 35 degrees C and to larvae at 20 degrees C. Piperonyl butoxide (PBO), a synergist, increased the relative potency of imidacloprid (1:5 imidacloprid:PBO) 16-fold at 26 degrees C against adults, but had no effect at 35 degrees C. No synergism occurred in larvae at 20 degrees C, but addition of PBO (1:5 imidacloprid:PBO) doubled toxicity at 26 degrees C. PBO (1:5 imidacloprid:PBO) could possibly be used to synergize imidacloprid premise treatments (20-30 degrees C), but it is not likely to be effective in pet treatments because no synergism occurred in adult fleas at 35 degrees C (average fur temperature of tested cats and dogs).