In addition, hourly health observations were conducted for 4 h following treatment with afoxolaner on Day 0. For both studies, seven days prior to treatment (designated as to Day 0) dogs were infested with 50 adult ticks of approximately equivalent sex ratio, which were removed and counted 48 h later. The dogs were ranked in order of these pre-treatment

tick counts (highest to lowest). The first two dogs were assigned to Block 1, the next click here two dogs to Block 2 and so on until 10 blocks of two dogs each were formed. Within blocks, dogs were randomly assigned to one of two treatment groups. Dogs in Group 1 were untreated controls. Dogs in Group 2 were treated once orally with the appropriate combination of soft chewables containing afoxolaner. Two sizes of chews were used: 0.5 g and 1.25 g, containing 11.3 mg and 28.3 mg of afoxolaner, respectively. The soft chewables are not designed to be divided, therefore, the dosing was administered as closely as possible to the minimum effective dose of 2.5 mg/kg using whole chews. The doses administered to dogs ranged from 2.57 to 3.96 mg/kg body weight in Study 1 and from 2.97 to 3.70 mg/kg body weight in Ribociclib manufacturer Study 2. The dogs

were observed during the 4 h following their treatment and daily throughout the study. Dogs were infested with 50 adult ticks (25 females and 25 males) on the day prior to treatment (Day – 1) and on Days 7, 14, 21, and 28. Forty-eight hours after treatment and 48 h after each of the subsequent re-infestations, ticks were removed and live ticks

were counted. These counts were conducted using a procedure involving Thymidine kinase methodical examination of all body areas using finger tips and/or a coarse tooth comb to sort through the hair and locate all ticks following WAAVP guidelines (Marchiondo et al., 2013). The two studies used unfed adult D. variabilis ticks from two separate laboratory-maintained populations. Each laboratory population had been established from ticks collected in the USA. Personnel responsible for collection of animal health and efficacy data were blinded to the treatment groups. Total counts of live ticks were transformed to the natural logarithm of (count + 1) for calculation of geometric means by treatment group at each time point. Percent reduction from the control group mean was calculated for the treated group at each post-treatment time point using the formula [(C − T)/C] × 100, where C is the geometric mean for the control group and T is the geometric mean for the treated group. The log counts of the treated group were compared to the log counts of the untreated control group using an F-test adjusted for the allocation blocks used to randomize the animals to the treatment groups. The comparisons were performed using a two-sided test with a 5% significance level.