James S. Hunter

THERAPEUTIC AND PERSISTENT EFFICACY OF FIPRONIL AGAINST BOOPHILUS MICROPLUS (ACARI : IXODIDAE) ON CATTLE

Cattle infested with all parasitic life-stages of Boophilus microplus (Canestrini) were treated once with a pour-on formulation of fipronil at 0.25, 0.5, or 1.0% active ingredient (AI) to determine the therapeutic efficacy. Additionally, cattle were infested at weekly intervals for 13 consecutive weeks after treatment to determine the persistent effectiveness of the three concentrations. Results of therapeutic efficacy of fipronil showed that with each increase in concentration there was a corresponding and often significant (P < 0.05) decrease in the numbers, fecundity and fertility of engorged females. The overall control of the Index of Reproduction (IR) achieved at 0.25 and 0.5% AI was 86.2 and 94.3%, respectively, while the control of the IR at the 1.0% AI concentration was 99.7%. The persistent efficacy of fipronil, like the therapeutic efficacy, was also dose related. The 0.25% AI treatment failed to provide complete protection (100%) against larval reinfestation, although control of the IR remained high (> 95%) for the first 6 weeks after treatment. In contrast, at 0.5% AI the control of the IR remained at 100% for the first 4 weeks after treatment, and was > 90% for an additional 2 weeks (6 weeks posttreatment). The 1.0% AI concentration afforded the longest residual activity, providing virtually 100% protection against larval reinfestation for 8 weeks after treatment was applied. Based on the results, neither the 0.25 or 0.5% AI concentration completely eliminated the immature stages of the tick (nymphs and larvae) on the host at the time of treatment (therapeutic efficacy), and the residual activity (persistent efficacy) did not prevent reinfestation by larvae for long periods after treatment. While the 1.0% AI concentration provided both therapeutic and residual efficacy that would be suitable for use in the eradication program, caution is still warranted because a few engorged females detached for up to 25 days after treatment, although none reproduced after the 6th day.

Evaluation of the curative and preventive efficacy of a single oral administration of afoxolaner against cat flea Ctenocephalides felis infestations on dogs

The efficacy of orally administered afoxolaner for treatment and prevention of repeated infestations with adult Ctenocephalides felis on dogs was evaluated in two studies after administration of a beef-flavored soft chew. In each study, 32 dogs were divided randomly into four equal groups. Dogs in Groups 1 and 3 were not treated and served as controls. Dogs in Groups 2 and 4 were treated on Day 0 with a combination of chewable tablets to be as close as possible to the minimum therapeutic dose of 2.5mg/kg. All animals were infested experimentally with unfed C. felis (100 ± 5) on Days -1, 7, 14, 21, 28 and 35. Flea killing efficacy was evaluated in both studies while, efficacy against flea egg production was assessed in Study 1. Live fleas were counted at 12 (Groups 1 and 2) and 24h (Groups 3 and 4), after treatment or after weekly infestations. In Study 1, flea eggs were collected and counted at either 12 or 24h after each flea infestation on Days 7, 14, 21, 28 and 35. The results of both studies demonstrate the long lasting and rapid efficacy of afoxolaner against C. felis, when administered as a single oral dose to dogs. For flea counts conducted 24h after treatment or infestation, efficacy was 100% for all time points up to Day 36 in both studies, except for one time point (99.9% on Day 22) for Study 2. For flea counts performed 12h after treatment or infestation, efficacy was ≥ 95.2% until Day 21 in both studies. Efficacy at 12h was ≥ 93.0% on Day 35 in Study 1 and ≥ 89.7% on Day 35 in Study 2. The treated groups had significantly fewer fleas than untreated control dogs in both studies for all flea counts (p=0.003 Study 1, p=0.0006 Study 2). In Study 1, for all egg counts performed at or beyond Day 7, efficacy in egg reduction was >99% for all time points between Days 7 and 35.

Onset and duration of immunity to equine influenza virus resulting from canarypox-vectored (ALVAC) vaccination.

Equine influenza virus remains an important problem in horses despite extensive use of vaccination. Efficacy of equine influenza vaccination depends on the onset and duration of protective immunity, and appropriate strain specificity of the immune response. This study was designed to test the protective immunity resulting from vaccination with the North American commercial ALVAC equine influenza vaccine (RECOMBITEK Influenza, Merial, USA)(1) against challenge with American lineage influenza viruses. In experiment 1, 12 ponies were vaccinated twice, at a 35 day interval, using the ALVAC-influenza vaccine expressing the HA genes of influenza A/eq/Newmarket/2/93 and A/eq/Kentucky/94 (H3N8), and 11 ponies served as unvaccinated controls. Six months after the second vaccination, all ponies were challenged with A/eq/Kentucky/91. In experiment 2, 10 ponies received one dose of the ALVAC-influenza vaccine, 10 ponies served as unvaccinated controls, and all ponies were challenge infected with A/equine/Ohio/03, 14 days after vaccination. Parameters studied included serological responses, and clinical disease and nasal viral shedding following challenge infection. In experiment 1, following the two-dose regimen, vaccinated ponies generated high titered anti-influenza virus IgGa and IgGb antibody responses to vaccination and demonstrated statistically significant clinical and virological protection to challenge infection compared to controls. Infection with A/eq/Kentucky/91 produced unusually severe signs in ponies in the control group, requiring therapy with NSAIDs and antibiotics, and leading to the euthanasia of one pony. In experiment 2 following the one-dose regimen, vaccinates generated IgGa responses pre-challenge, and anamnestic IgGa and IgGb responses after challenge. Vaccinates demonstrated statistically significant clinical and virological protection to challenge infection compared to controls. The results of this study clearly demonstrate the early onset, and 6-month duration of protective immunity resulting from ALVAC-influenza vaccination against challenge with American lineage equine influenza viruses.

Efficacy of a novel topical combination of fipronil, amitraz and (S)-methoprene for treatment and control of induced infestations of brown dog ticks (Rhipicephalus sanguineus) on dogs.

Four laboratory studies were conducted to demonstrate that a single topical dose of a novel spot-on combination containing fipronil, amitraz and (S)-methoprene (CERTIFECT™, Merial Limited, GA, USA) is efficacious against the brown dog tick, Rhipicephalus sanguineus. In each study, 6-8 male and 6-8 female purpose-bred, laboratory mongrels, terrier cross or Beagles were randomly assigned to one of two study groups (treated and untreated), based on pre-treatment parasite counts. Starting on the day before treatment, each dog was infested weekly with 50 ticks. Ticks were thumb counted at various time points after treatment and weekly infestations starting as early as 6h and continued at 12, 18 and 24h depending on the study. Ticks were removed and counted at 48 h after treatment and weekly infestations. CERTIFECT provided rapid and excellent control of pre-existing and newly acquired infestations of R. sanguineus with efficacy as high as 93% within the first 12h after a single topical treatment. Excellent control (>96%) of R. sanguineus as early as 18 h, following post treatment infestations was maintained for at least 35 days.

The prevention of attachment and the detachment effects of a novel combination of fipronil, amitraz and (S)-methoprene for Rhipicephalus sanguineus and Dermacentor variabilis on dogs

A novel combination of fipronil, amitraz and (S)-methoprene (CERTIFECT™, Merial Limited, GA, USA) was evaluated for the prevention of attachment of ticks and its ability to cause detachment of ticks. For the two prevention of attachment studies, 20 purpose-bred beagles were allocated each to two equal groups based on pretreatment tick counts (treated and untreated). Each dog was exposed to 50 adult Rhipicephalus sanguineus and Dermacentor variabilis weekly starting 24h after treatment. In study 1 infestations with R. sanguineus were discontinued after Day 7 but continued to Day 28 for D. variabilis in both studies. Counts of ticks by species were made 2, 4 and 24h after exposure to ticks. Ticks not attaching to dogs were evaluated for viability. For the evaluation of detachment study, 16 purpose-bred beagles were allocated each to two equal groups based on pretreatment tick counts (treated and untreated). Each dog was infested with 50 unfed R. sanguineus and D. variabilis adults on Day -2. Ticks were thumb counted without removal on all dogs on Day -1, and at 4, 12, and 24h after treatment. Ticks were counted and removed at 48 h after treatment. Dogs treated with the novel combination had significantly (p<0.05) lower total numbers of attached R. sanguineus and D. variabilis than untreated controls at 4h through Day 7. For R. sanguineus, percent reduction of attachment at 24h after infestation through Day 29 ranged from 94.5% to 100%. For D. variabilis, the percent reduction of attachment at 24h through Day 22 was above 98.0%. These studies demonstrate that novel combination can disrupt attachment of R. sanguineus and D. variabilis for up to 28 days following treatment. Of those ticks that are exposed to the treatment, even if they do not attach to the dog and remain in the environment, greater than 90% (p<0.05) die within 24h for 2-3 weeks following treatment. Also, for those dogs infested with ticks at the time of treatment, the novel combination causes significant detachment (p<.05) starting at 12h and reaching 98.9% by 48 h after treatment. This product provides an effective means for controlling ticks infesting dogs and limiting the spread of tick transmitted diseases. Additionally, the mortality of ticks exposed to CERTIFECT will reduce infestation of the dogs environment.

Efficacy of a novel topical combination of fipronil, amitraz and (S)-methoprene for treatment and control of induced infestations with four North American tick species (Dermacentor variabilis, Ixodes scapularis, Amblyomma americanum and Amblyomma maculatum) on dogs

Five laboratory studies were conducted to confirm that a single topical dose of the novel combination of fipronil, amitraz and (S)-methoprene, CERTIFECT™ (Merial Limited, GA, USA), is efficacious for the rapid control of pre-existing infestations and the prevention of new infestations with Ixodes scapularis, Dermacentor variabilis, Amblyomma americanum and Amblyomma maculatum for at least 28 days on dogs. In each study, 8 male and 8 female purpose-bred, laboratory beagles were randomly assigned to one of two study groups (treated and untreated). Starting on the day before treatment, each dog was infested weekly with about 30 or 50 ticks, depending on the study. Treatment with the novel combination rapidly eliminated pre-existing infestations and controlled weekly re-infestations for at least 28 days. Pre-existing infestations with all four tick species were rapidly and effectively reduced, with post-treatment therapeutic efficacies ranging from 91.7 to 99.5% within 18-48 h post treatment. Amblyomma maculatum numbers were significantly (p<0.05) reduced on treated dogs from the first tick counts as early as 6h post-treatment. All subsequent infestations with each of the 4 tick species were quickly disrupted, with prophylactic efficacies greater than 90% within 18-48 h post-infestation for at least a full month. Because the combination of fipronil, amitraz and (S)-methoprene quickly starts disrupting and killing ixodid ticks within hours of treatment, with similar high levels of efficacy maintained for at least 28 days in these and other studies, the authors conclude that a single topical treatment with CERTIFECT may prevent the transmission of most infectious agents carried by ixodid ticks for at least one month.

The efficacy of eprinomectin extended-release injection against naturally acquired nematode parasites of cattle, with special regard to inhibited fourth-stage Ostertagia larvae

The efficacy of eprinomectin in an extended-release injection (ERI) formulation in the treatment of cattle harboring naturally acquired nematode populations (including inhibited nematodes) was evaluated. Five studies were conducted under a similar protocol in the USA, the UK, and in Germany. All study animals were infected by grazing naturally contaminated pastures. The adequacy of pasture infectivity was confirmed by examining tracer calves prior to allocation and treatment of the study animals. The cattle were of various breeds or crosses, weighing 79-491 kg, and aged approximately 6-15 months. In each study, 20 animals were infected by grazing, and then removed from pasture and housed in a manner to preclude further nematode infections for 8-16 days until treatment. Animals were blocked based on descending pre-treatment body weight and randomly allocated to one of two treatments: ERI vehicle (control) at 1 mL/50 kg body weight or eprinomectin 5% (w/v) ERI at 1 mL/50 kg body weight (1.0 mg eprinomectin/kg). Treatments were administered once on Day 0 by subcutaneous injection in front of the shoulder. For parasite recovery and count, all study animals were humanely euthanized 14/15 days after treatment. Cattle treated with eprinomectin ERI had significantly (p<0.05) fewer of the following nematodes than the controls with overall reduction of parasite counts of ≥94%: adult Dictyocaulus viviparus, Capillaria spp., Cooperia oncophora, Cooperia pectinata, Cooperia punctata, Cooperia surnabada, Haemonchus placei, Nematodirus helvetianus, Oesophagostomum radiatum, Ostertagia lyrata, Ostertagia ostertagi, Trichostrongylus axei, Trichostrongylus colubriformis, Trichuris discolor, Trichuris skrjabini, and Trichuris spp.; developing fourth-stage larvae of Ostertagia spp. and Trichostrongylus spp.; and inhibited fourth-stage larvae of Cooperia spp., Haemonchus spp., Nematodirus spp., Oesophagostomum spp., Ostertagia spp., and Trichostrongylus spp. Animal treatments were well accepted, with no adverse reactions to treatment observed in any study animals. The results of this series of controlled studies demonstrated high therapeutic efficacy and acceptability of eprinomectin ERI against pulmonary nematodes and a wide range of gastrointestinal parasitic infections, including inhibited gastrointestinal nematodes, in cattle.