Patricia A. Payne

Development of a Larval Bioassay for Susceptibility of Cat Fleas (Siphonaptera: Pulicidae) to Imidacloprid

Abstract Strategies for controlling cat fleas, Ctenocephalides felis felis (Bouché), have undergone dramatic changes in the past 5 yr. With the advent of on-animal treatments with residual activity the potential for the development of insecticide resistance increases. A larval bioassay was developed to determine the baseline susceptibility of field-collected strains of cat fleas to imidacloprid. All four laboratory strains tested showed a similar level of susceptibility to imidacloprid. Advantages of this bioassay are that smaller numbers of fleas are required because flea eggs are collected for the test. Insect growth regulators and other novel insecticides can also be evaluated. Using a discriminating dose, the detection of reduced susceptibility in field strains can be determined with as few as 40 eggs.

Effect of 0.29% w/w fipronil spray on adult flea mortality and egg production of three different cat flea, Ctenocephalides felis (Bouché), strains infesting cats

To evaluate the effect of fipronil spray on adult flea mortality and flea egg production of three different cat flea, Ctenocephalides felis (Bouché) strains, 30 domestic short hair cats were randomly allocated into six groups of five cats each. On day 0, cats in groups 2, 4 and 6 were treated with fipronil at 5-6ml/kg. Cats in groups 1, 3 and 5 served as untreated controls. On days -2, 7, 14, 21, and 28 each cat was infested with 50 adult cat fleas. Groups 1 and 2 were infested with fleas from the Kansas1 Colony (KS1) strain. Groups 3 and 4 were infested with a recently colonized cat flea strain from Florida (R6). Groups 5 and 6 were infested with fleas from the ARC strain. The adulticidal activity of fipronil was determined by flea comb counts 48h after treatment and then 48h after each reinfestation. Any flea eggs produced during the infestations were collected and counted prior to the 48h comb counts. Fipronil spray was > or = 99.5% effective against adults of all three cat flea strains when applied during an active infestation. Fipronil spray provided > or = 98.2 and > or = 99.5% control of adult fleas and egg production, respectively, for all strains through week 2. On days 23 and 30 control of R6 adults and egg production was significantly lower than either the ARC or the KS1 strain. On day 30, control of R6 adults and egg production was 77.3 and 87.3%, respectively. Control of KS1 adults and egg production on day 30 was significantly lower than the ARC strain. Fipronil provided > or = 99.5 and > or = 99.9% control of ARC fleas and egg production, respectively, throughout the entire study. The susceptibility to fipronil for the three strains was also evaluated on filter paper pesticide bioassays. The R6 strain was found to be less susceptible than the KS1 and ARC strains. The LC(95) estimates for the strains were 10.13, 4.77 and 2.62mg/m(2) for the R6, ARC and KS1 strains, respectively.

Efficacy of indoxacarb applied to cats against the adult cat flea, Ctenocephalides felis, flea eggs and adult flea emergence

BackgroundA study was conducted to evaluate the effect of indoxacarb applied to cats on adult cat fleas, Ctenocephalides felis, flea egg production and adult flea emergence.MethodsSixteen cats were selected for the study and allocated to two treatment groups. Eight cats were treated with a 19.5% w/v topical spot-on solution of indoxacarb on day 0 and eight cats served as untreated controls. Each cat was infested with 50 fleas on Days -2, 7, 14, 21, 28, 35 and 42. On Days 1, 2, and 3, and at 2 and 3 days after each post treatment reinfestation flea eggs were collected from the pan under each cat cage. Eggs were counted and viability assessed by evaluating adult flea emergence 28 days after egg collection. Three days after treatment or infestation, each cat was combed to remove and count live fleas.ResultsTreatment with indoxacarb provided 100% efficacy following infestations on day -2, 7, 14, 21 and 28 and efficacy was 99.6% following infestations on days 35 and 42. Egg production from indoxacarb treated cats was reduced by 99.9% within 72 hours of treatment. For subsequent infestations no eggs were produced from treated cats from day 8 through day 30. Egg production was still reduced by ≥95.8% through day 45. Indoxacarb treatment also reduced adult flea emergence from eggs for 5 weeks after treatment. The combination of reduction in egg numbers and egg viability from indoxacarb treated cats reduced predicted flea emergence by 100% from days 2 – 31 and 99.9%, 100%, 96.4% and 99.0% on days 37, 38, 44 and 45, respectively.ConclusionsA topical spot-on formulation of indoxacarb provided ≥99.6% efficacy against flea infestations on cats for 6 weeks following a single treatment. Indoxacarb also eliminated or markedly reduced egg production for the entire evaluation period and reduced the viability of the few eggs that were produced from Day 1 through Day 38. Given indoxacarb’s effect on adult fleas, egg production and egg viability; this formulation can interrupt flea reproduction on treated cats for at least 6 weeks after treatment.

Determining a Diagnostic Dose for Imidacloprid Susceptibility Testing of Field-Collected Isolates of Cat Fleas (Siphonaptera: Pulicidae)

Abstract The susceptibility of four laboratory strains of cat fleas, Ctenocephalides felis (Bouché), to imidacloprid was determined by three different laboratories, by using a standardized bioassay protocol. The probit lines generated by the different laboratories were very similar, with LC50 values ranging from 0.32 to 0.81 ppm. Based on these data, a diagnostic dose (DD) of 3 ppm imidacloprid in larval rearing media was provisionally identified for detecting shifts in tolerance, possibly as a consequence of incipient imidacloprid resistance. None of the larvae from the susceptible laboratory strains survived the DD. Eighteen field-collected isolates were evaluated for their susceptibility to imidacloprid and to validate a DD of 3 ppm. Probit lines from 18 field-collected isolates were very similar, with LC50 values ranging from 0.14 to 1.52 ppm. When exposed to the DD, between 3 and 10% of the exposed larvae emerged as adults from only three of the 18 isolates. All other field isolates gave 100% mortality at the DD. Under the criteria established (>5% survivorship at 3 ppm), two isolates would be established on mammalian hosts and more extensive tests conducted to exclude or confirm the presence of resistance. The DD of 3 ppm is robust enough to eliminate most of the susceptible isolates collected until today, yet low enough to identify possible isolates for further testing.

Efficacy of selamectin, spinosad, and spinosad/milbemycin oxime against the KS1 Ctenocephalides felis flea strain infesting dogs.

BackgroundA study was conducted to evaluate and compare the efficacy of selamectin, spinosad, and spinosad/milbemycin oxime against the KS1 strain of Ctenocephalides felis on dogs.MethodsForty-eight dogs were selected for the study and two batches of 24 were blocked and allocated randomly to treatment groups and flea count times. There were four treatment groups of 12 dogs each: negative control, topical selamectin, oral spinosad/milbemycin oxime, and oral spinosad. Each dog was infested with 100 fleas on Days -2, 7, 14, 21 and 28. Within each treatment group, six dogs were flea counted at 24 hours and six at 48 hours after treatment or post-infestation. On Day 0, dogs received a single treatment of the appropriate drug according to the approved commercial label.ResultsEfficacy of selamectin against an existing flea infestation was 60.4% and 91.4% at 24 and 48 hours, respectively, whereas spinosad/milbemycin oxime and spinosad were 100% at both time points. All products were >90% effective within 24 hours after subsequent infestations on Days 7, 14 and 21. Following the Day 28 flea infestation, selamectin was 93% and 95.7% effective at 24 and 48 hours, respectively. Whereas the efficacy of spinosad/milbemycin oxime following the day 28 infestation was 84.7% and 87.5% at 24 and 48 hours, respectively and spinosad alone was 72.9% and 76.3% effective at 24 and 48 hours, respectively.ConclusionsAfter initial application, the two oral spinosad products had a more rapid onset of flea kill than topical selamectin which took up to 48 hours to control (>90%) the existing infestation. However, for subsequent weekly flea infestations selamectin had similar or better efficacy than spinosad or spinosad/milbemycin oxime at 24 and 48 hours after infestation. Spinosad/milbemycin oxime and spinosad were >90% effective against the KS1 strain from Day 1 to Day 23. Whereas, selamectin was >90% effective against the KS1 strain of C. felis from Day 2 to Day 30.

American Association of Veterinary Parasitologists’ review of veterinary fecal flotation methods and factors influencing their accuracy and use—Is there really one best technique?

The principle of fecal flotation is based on the ability of a solution to allow less dense material (including parasite elements) to rise to the top. However, there are numerous factors that will influence the accuracy and use of such a theoretically simple technique. Whether or not centrifugation is used appears to have an impact on the ability to detect some parasites, but not others. Using a flotation solution with a relatively high specific gravity favors the simultaneous flotation of the diagnostic stages of many different parasites while, at the same time, making recognition of some more difficult because of distortion as well as the amount of debris in the preparation. Dilution methods tend to be less accurate because they require extrapolation; however, they are quicker to perform, in part, because of the cleaner preparation. Timing is a critical factor in the success of all flotation methods, as is technical ability of the personnel involved. Thus, simplicity, low costs and time savings have generally favored gravitational flotation techniques (including the McMaster technique and its modifications). How accurate the method needs to be is dependent upon the purpose of its use and choice of method requires an understanding of analytical sensitivity and expected levels of egg excretion. In some instances where the difference between, for example, 0 and 50 eggs per gram is insignificant with regards to management decisions, less accurate methods will suffice. In others, where the presence of a parasite means treatment of the animal regardless of the numbers of eggs present, methods with higher analytical sensitivities will be required, particularly for those parasites that pass few eggs. For other uses, such as the Fecal Egg Count Reduction Test, accuracy may become critical. Therefore, even though recommendations for standardized fecal flotation procedures have been promoted in the past, it is clear that the factors are too numerous to allow for the recommendation of one, or even a few, procedures for all purposes.

Evaluation of indoxacarb and fipronil (s)-methoprene topical spot-on formulations to control flea populations in naturally infested dogs and cats in private residences in Tampa FL. USA.

BackgroundA study was conducted to evaluate and compare the effectiveness of two different spot-on topical flea products to control flea infestations on naturally infested dogs and cats in Tampa, FL USA.MethodsThirty-two dogs and 3 cats with natural flea infestations living in 18 homes were treated topically with a 19.53% w/w spot-on formulation of indoxacarb. Another thirty dogs and 2 cats living in 19 different homes were treated topically with either fipronil (9.8% w/w)/(s)-methoprene (8.89% w/w) or fipronil (9.8% w/w)/(s)-methoprene (11.8% w/w), respectively. All products were applied according to label directions by study investigators on day 0 and again between days 28 and 30. Flea populations on pets were assessed using visual area counts and premise flea infestations were assessed using intermittent-light flea traps on days 0, 7, 14, 21, 28–30, 40–45, and 54–60.ResultsA single application of the indoxacarb or fipronil (s)-methoprene formulations reduced flea populations on pets by 97.8% and 85.5%, respectively within 7 days. One month (28–30 days) after treatment the indoxacarb and fipronil (s)-methoprene formulations reduced on-animal flea burdens by 95.0% and 49.5%, respectively. Following two monthly applications of either the indoxacarb or fipronil (s)-methoprene formulations, pet flea burdens were reduced by 99.1% and 54.8%, respectively, by days 54 – 60. At the end of the two month study, 77.1% and 15.6% of the dogs and cats in the indoxacarb and fipronil (s)-methoprene treatment groups, respectively were flea free. Flea numbers in the indoor-premises were markedly reduced in both treatment groups by days 54–60, with 97.7% and 84.6% reductions in intermittent-light flea trap counts in the indoxacarb and fipronil (s)-methoprene treatment groups, respectively.ConclusionsThis in-home investigation conducted during the summer of 2013 in subtropical Tampa, FL, is the first published U.S field investigation of the indoxacarb topical formulation. The indoxacarb formulation was able to effectively control flea populations in heavily flea infested pets and homes. The efficacy achieved by the fipronil (s)-methoprene formulation against flea infestations on these pets was lower than in previous investigations using the same study design.

PROGRESS OF THE INTERNATIONAL WORK OF THE ''IMIDACLOPRID FLEA SUSCEPTIBILITY MONITORING TEAM''

The “Imidacloprid Flea Susceptibility Monitoring Team” has aim to develop and validate a bioassay to effectively monitor and document susceptibility of cat flea (Ctenocephalides felis) isolates to imidacloprid. A larval bioassay was developed, standardized and validated and agreed upon by the team as the reference diagnostic test kit as research has shown that the proposed WHO adult test was not reliable. The selected 3 ppm discriminating dose, determined from evaluating year 2000 field isolates, was approximately 2 times the highest LC95 of the control laboratory flea strains. In 2001 and 2002, this standardized bioassay was used to test more than 190 separate egg collections from individual flea isolates from USA, UK and Germany. If survivorship in the 3 ppm assay is confirmed, the LD50 values of this isolate and the laboratory strains will be determined by a dose-response study in the range of 0.005 to 3 ppm imidacloprid. By comparison of these LD50 values it can be estimated whether a shift in susceptibility to imidacloprid has occurred. In either case the flea isolate will be researched extensively. Currently an in-vivo test which will evaluate the on-host efficacy of Advantage® is being established. None of the to date tested isolates revealed reduced susceptibility to imidacloprid.

Strategic use of ivermectin during pregnancy to control toxocara canis in greyhound puppies.

Twenty-one greyhound bitches were bred (Day 0) and housed throughout their pregnancies on three greyhound breeding farms in Kansas. These dogs were assigned randomly to one of four treatment groups. Group A dogs (6) were given ivermectin subcutaneously (300 microg/kg) on Day 0 (the first day the dogs were bred), and Days 30 and 60 of gestation. Group B dogs (6) were given ivermectin (300 microg/kg SQ) on Day 42. Group C dogs (3) were given ivermectin (300 microg/kg SQ) on Days 0, 30, and 60 plus 10 days after whelping. Group D dogs (6) served as controls and received no anthelmintic. Bitches and puppies were moved to the university on the day after birth and were maintained inside for 28 days. Weekly quantitative fecal exams were done on the bitches during this time. The puppies were euthanized humanely at 28 days of age. Intestinal parasites were recovered, identified, counted, sexed, and preserved in either 10% formalin or frozen at -70 degrees C. The geometric mean numbers of adult Toxocara canis in the small intestines for Group A puppies (n = 40) were 2.8, 8.5 for Group B puppies (n = 39), and 29.7 for Group D puppies (n = 28). No adults were found in the Group C puppies (n = 15). The geometric mean eggs per gram of feces from the pups in group A, B, and D were 1.3, 704, and 27, 134, respectively. No eggs were recovered from the Group C pups. The strategic use of ivermectin at 300 microg/kg in greyhound bitches on Days 0, 30, and 60 of gestation reduced the worm burden carried by the puppies by 90% and the actual number of eggs passed into the environment by 99.8%. The same dose on day 42 reduced the worm burden by 71.4% and the number of eggs passed into the environment by 97.4%. This dose given on days 0, 30, and 60 plus 10 days postwhelping, reduced the worm burden by 100%, and no eggs were passed into the environment.

Efficacy of imidacloprid + moxidectin and selamectin topical solutions against the KS1 Ctenocephalides felis flea strain infesting cats

BackgroundTwo studies were conducted to evaluate and compare the efficacy of imidacloprid + moxidectin and selamectin topical solutions against the KS1 flea strain infesting cats. In both studies the treatment groups were comprised of non-treated controls, 6% w/v selamectin (Revolution®; Pfizer Animal Health) topical solution and 10% w/v imidacloprid + 1% w/v moxidectin (Advantage Multi® for Cats, Bayer Animal Health) topical solution. All cats were infested with 100 fleas on Days -2, 7, 14, 21, and 28. The difference in the studies was that in study #1 efficacy evaluations were conducted at 24 and 48 hours post-treatment or post-infestation, and in study #2 evaluations were conducted at 12 and 24 hours.ResultsIn study #1 imidacloprid + moxidectin and the selamectin formulation provided 99.8% and 99.0% efficacy at 24 hours post-treatment. On day 28, the 24 hour efficacy of the selamectin formulation dropped to 87.1%, whereas the imidacloprid + moxidectin formulation provided 98.9% efficacy. At the 48 hour assessments following the 28 day infestations, efficacy of the imidacloprid + moxidectin and selamectin formulations was 96.8% and 98.3% respectively. In study # 2 the efficacy of the imidacloprid + moxidectin and selamectin formulations 12 hours after treatment was 100% and 69.4%, respectively. On day 28, efficacy of the imidacloprid + moxidectin and selamectin formulations 12 hours after infestation was 90.2% and 57.3%, respectively. In study #2 both formulations provided high levels of efficacy at the 24 hour post-infestation assessments, with selamectin and imidacloprid + moxidectin providing 95.3% and 97.5% efficacy, following infestations on day 28.ConclusionsAt the 24 and 48 hour residual efficacy assessments, the imidacloprid + moxidectin and selamectin formulations were similarly highly efficacious. However, the imidacloprid + moxidectin formulation provided a significantly higher rate of flea kill against the KS1 flea strain infesting cats at every 12 hour post-infestation residual efficacy assessment. Both formulations should provide excellent flea control for an entire month on cats.