Michelle Zakson-Aiken

Systemic Activity of the Avermectins Against the Cat Flea (Siphonaptera: Pulicidae)

Abstract Ivermectin has potent systemic activity against numerous species of nematodes and arthropods, but there are some important species in these two groups, such as the cat flea, Ctenocephalides felis (Bouché), that appear to be refractory to it. In an effort to determine if the lack of systemic activity against C. felis is specific to ivermectin, or if it is a class-wide phenomenon, 20 avermectin derivatives were tested in an artificial membrane flea feeding system at concentrations of 20, 10, and 1 μg/ml. Results showed that ivermectin had LC90 and LC50 values against fleas of 19.1 and 9.9 μg/ml, respectively. Only four of the other 19 compounds evaluated possessed both LC90 and LC50 values more potent than ivermectin and even then the advantage was modest. Among those four compounds was a two-fold increase in potency relative to ivermectin when the LC90 values were considered (range, 9.2–10.3 μg/ml) and a two- to eight-fold increase when the LC50 values were examined (range, 1.23–5.26 μg/ml). Neither the possession nor the number of oleandrosyl sugars on the macrocyclic backbone were relevant for additional flea activity because among these four compounds were two disaccharides, a monosaccharide and an aglycone. Also, bond disposition between C-22 and 23 did not contribute to increase in activity because these molecules comprise members with either single or double bonds. One of these avermectin analogs was scaled-up and tested subcutaneously in a dog at >100 times the commercial ivermectin dosage and zero efficacy was observed against the flea. We conclude that even the best in vitro avermectin does not have the in vivo potential to become a commercial oral or subcutaneous flea treatment for companion animals.

Development of a mouse model to determine the systemic activity of potential flea-control compounds.

Probe studies were performed to determine if the cat flea (Ctenocephalides felis), the most common ectoparasite of companion animals, will feed on laboratory mice and, if so, to incorporate this into a small animal assay to detect systemically active compounds. Consequently, a protocol was developed which incorporated acepromazine maleate to temporarily sedate various strains of mice and allow fleas a window of time to feed undisturbed. For validation of the model, CD-1 mice were dosed per os with seven known insecticides at 30, 10 and 1mg/kg. Mice were sedated with 0.0125 ml acepromazine maleate intraperitoneally, and infested with fleas. After 2h, fleas were removed, one-third were examined immediately to confirm the occurrence of feeding, and 77% were found to have ingested a blood meal. The remaining fleas were incubated for 24h to determine mortality. Nitenpyram, the active ingredient in Capstar, was highly active (>94%) at 1mg/kg. Selamectin, the active ingredient in Revolution, was very active (86%) at 10mg/kg, but inactive at 1mg/kg. Fipronil, the active ingredient of Frontline Topspot, was very active (83%) at 30 mg/kg, moderately active (54%) at 10mg/kg and inactive at 1mg/kg. Cythioate, the active ingredient in Proban, and nodulisporic acid, a recently discovered oral insecticide, were moderately active (64 and 55%, respectively) at 10mg/kg, but both were inactive at 1mg/kg. Lufenuron and ivermectin exhibited no efficacy at any level tested. These findings suggest that this mouse model can effectively identify systemic flea-control leads and, subsequently, reduce the use of large animals in research.

Development of an Assay for the Screening of Compounds Against Larvae of the Cat Flea (Siphonaptera: Pulicidae)

Abstract Larvae of the cat flea,Ctenocephalides felis(Bouché), are the target of numerous growth regulators. This study explores the development of an assay that tests the susceptibility of cat flea larvae to a wide range of compounds. Different rearing media and containment units were tested that would facilitate optimization. Larvae of various ages were compared, and 7-d-old larvae were found to be optimal because they were the most uniform in size and age and exhibited a need to feed. The assay could be used to distinguish insecticides from growth inhibitors. The insecticides chlorpyrifos and carbaryl caused 100% larval mortality in 24 h at 10 ppm, and cythioate and fipronil killed the larvae at concentrations of ≥100 ppm within 24 h. The insect growth regulators methoprene and pyriproxifen caused molt delay at concentrations of 100 ppm and bioallethrin delayed molt at 1,000 ppm. This assay can be used to identify compounds that are specific to cat flea larvae that may be useful in the control of cat flea infestations.