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Abstract Background Credelio™ (lotilaner; Elanco) is indicated for the treatment of flea and tick infestations on cats at a recommended lotilaner dose rate of 6–24 mg/kg. This study evaluated the efficacy and safety of lotilaner following a single oral administration to cats for the treatment and prevention of adult Ctenocephalides felis fleas and flea egg production under laboratory conditions. Methods Two treatment groups of ten cats each were used in this study. One group was treated with lotilaner at a dose rate of 6−9 mg/kg on Day 0 and the other group served as the control group. Each cat was infested with 100 unfed adult fleas on days –1, 6, 13, 20 and 29. At 24 h post-treatment or post-infestation, each cat was combed to remove and count adult live fleas. At each time point, flea eggs were also collected and counted from under each cat cage. Results Following a single oral administration of lotilaner at a minimum dose rate of 6 mg/kg (range 6.00−8.57 mg/kg), the lotilaner group displayed 100%, 100%, 99.9%, 99.9% and 99.8% efficacy against adult live flea counts as compared to the control group on Days 1, 7, 14, 21 and 30, respectively. At each time point, adult flea counts from the lotilaner-treated cats were significantly lower (P |