M. Pollmeier

The prevention of transmission of Babesia canis canis by Dermacentor reticulatus ticks to dogs using a novel combination of fipronil, amitraz and (S)-methoprene.

Four groups of seven dogs were treated topically with a novel combination of fipronil, amitraz and (S)-methoprene in a spot-on formulation (CERTIFECT™, Merial Limited, GA, USA) on 28, 21, 14 and 7 days prior to tick infestation, respectively and acaricidal efficacy and transmission blocking compared with an untreated control group (seven dogs). All dogs were infested with adult Dermacentor reticulatus ticks harbouring Babesia canis canis. Babesia canis canis was transmitted by D. reticulatus to all seven untreated control dogs, confirmed following demonstration of clinical signs, by the detection of B. canis parasites in thin blood smears and B. canis canis PCR-RLB DNA assay on blood and the development of B. canis canis antibody titres by 14-21 days after tick infestation. The majority of treated dogs remained sero-negative for 42 days after infestation. Therefore, the treatment of dogs with CERTIFECT applied up to 28 days prior to infestation with D. reticulatus harbouring B. canis canis, successfully prevented the development of clinical signs of canine babesiosis.

The ability of a topical novel combination of fipronil, amitraz and (S)-methoprene to protect dogs from Borrelia burgdorferi and Anaplasma phagocytophilum infections transmitted by Ixodes scapularis.

Healthy, purpose-bred laboratory beagle dogs that had not been exposed to ticks and were seronegative for Borrelia burgdorferi and Anaplasma phagocytophilum were randomly assigned to four groups of eight dogs each. Control group 1 was not treated. Groups 2, 3 and 4 were treated with a single topical application of a new formulation of fipronil, amitraz and (S)-methoprene (CERTIFECT™, Merial Limited, GA, USA) at 28, 21 or 14 days prior to tick infestation, respectively. Each dog was infested with 25 female and 25 male field-collected adult Ixodes scapularis ticks that had infection rates of 66% for B. burgdorferi sensu stricto and 23% for A. phagocytophilum, as determined by polymerase chain reaction. Two and five days after tick infestation, control dogs had an average of 9.5 and 13.9 attached adult female ticks, respectively, whilst the 24 treated dogs remained tick-free aside from a single tick on the 2nd day after infestation. Serial serological tests demonstrated that the ticks successfully infected 8/8 control dogs with B. burgdorferi and co-infected 6/8 with A. phagocytophilum. B. burgdorferi infection also was confirmed in most control dogs by culture (6/8) and PCR (7/8) of skin biopsies. In contrast, CERTIFECT protected all 24 treated dogs against infection by both B. burgdorferi and A. phagocytophilum, as demonstrated by their negative serological tests throughout the study and the absence of any positive skin biopsy culture or PCR in these dogs.

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.

Evaluation of the efficacy of fipronil formulations in the treatment and control of biting lice, Trichodectes canis (De Geer, 1778) on dogs

The efficacy of FRONTLINE SPRAY (0.25% (w/v) fipronil), FRONTLINE SPOT-ON FOR DOGS (10% (w/v) fipronil) and FRONTLINE PLUS FOR DOGS (10% (w/v) fipronil and 9% (S)-methoprene) against the biting louse Trichodectes canis on dogs was confirmed under laboratory conditions. A field study evaluated the efficacy of a single topical application of FRONTLINE SPRAY and FRONTLINE SPOT-ON against the parasite on dogs. A total of 48 dogs of mixed breeds, both sexes, aged 2 months-7 years and weighing 1.8-37.0kg were used. The animals were either experimentally (laboratory study) or naturally (field study) infested with lice. Dogs were housed individually in order to prevent contact between animals. In the laboratory study, animals were allocated based on pre-treatment louse counts from 38 hair coat-partings per animal. Dogs were randomly assigned to the four treatment groups: (1) untreated control; (2) FRONTLINE SPRAY, at 6ml/kg; (3) FRONTLINE SPOT-ON as per label and (4) FRONTLINE PLUS as per label. Dogs in treatment groups 2-4 were treated twice topically on Days 0 and 28. The number of live lice in the 38 hair coat-partings per animal were counted on Days 2, 7 and weekly to Day 63. In addition, a whole body comb count was performed on Day 63. No live T. canis were found on dogs treated with FRONTLINE formulations at any post-treatment examination. The difference from controls was significant (P<0.01) for each product at each examination. Based on the whole body comb count at Day 63, the efficacy of each product was determined to be 100%. In the field study, dogs were allocated in strict order of presentation. Dogs were randomly allocated to one of the three treatment groups: (1) BOLFO collar (propoxur); (2) FRONTLINE SPRAY, at 6ml/kg and (3) FRONTLINE SPOT-ON as per label. Dogs were treated once topically on Day 0. The number of live lice was determined by whole body searches on Days 0 (pre-treatment), 2, 28 and 42. Louse counts of dogs treated with either FRONTLINE SPRAY, or FRONTLINE SPOT-ON were not different than those of dogs receiving the propoxur collar. The efficacy was determined to be >98% on Day 2 and, 100% on Days 28 and 42 in all treatment groups. The results of these studies demonstrate that fipronil in topical formulations is effective for treatment and control of biting lice (T. canis) infestations on dogs.

Efficacy of eprinomectin against Linognathus vituli and Bovicola bovis on calves

THE long-nosed sucking louse Linognathus vituli and the biting louse Bovicola (Damalinia) bovis are frequently found on calves, especially during housing in winter and early spring. Cattle lice are considered by some cattle producers and veterinarians to be underestimated as a cause of economic losses. However, the losses in productivity due to louse infestation can be significant (Drummond and others 1988). Infestation with L vituli or B bovis causes discomfort to animals due to skin irritation, which results in higher rates of rubbing, self-licking and scratching (Weeks and others 1995). The ectoparasites may also damage an animal’s hide, as a result of mechanical injuries caused by the animal rubbing itself on equipment (Baker and Oormadzi 1978, Cooper 1993), and also by skin irritation reactions (Hendrikx and others 1988, Bugby and others 1990, Webster and Bugby 1990, Schröer 1992, Nafstad and Grønstøl 2001, Coles and others 2003); these effects can be economically important. As the damage to the skin only becomes visible after tanning and dyeing, the leather industry is affected (Webster and Bugby 1990, Schröer 1992, Cooper 1993). Recently, infestations with L vituli have been found to be associated with anaemia and even mortality in young calves (Otter and others 2003). Furthermore, lice may also contribute to the spread of dermatomycoses in cattle herds (Cicias̆vili 1969). The control of ectoparasites by the application of topical ivermectin may result in a significant improvement in leather quality (Halligan 1991, Titchener and Newbold 1992, Hendrikx 1996). Eprinomectin, a new member of the macrocyclic lactone family, has been developed in a pour-on formulation for the treatment of endoand ectoparasites of cattle of all classes, including lactating dairy cows (Shoop and others 1996a, b). This short communication describes the results of studies conducted in Europe to confirm the efficacy of the selected dose of eprinomectin against lice on housed calves. Three studies were conducted in the UK, Germany and Italy using topically applied eprinomectin against natural infestation of calves with L vituli or B bovis. All the animals were handled with due regard for their welfare and studies were conducted under the relevant Institutional Animal Care and Use Committee oversight. Details of the animals are given in Table 1. The calves were individually tethered (studies 1 and 2) or individually penned (study 3) to prevent self-grooming and grooming of or contact with other calves. They were allocated to one of two treatment groups by restricted randomisation based on lice counts (study 1) or on their pretreatment bodyweight (studies 2 and 3). In all studies one group formed a control group (untreated in study 1 or treated with vehicle [placebo] in studies 2 and 3) and the calves in the other group were treated with 1 ml of a 0·5 per cent eprinomectin solution (Eprinex Pour-On; Merial) per 10 kg bodyweight, that is, 500 μg eprinomectin/kg bodyweight. The vehicle was applied at the same volume rate (1 ml/10 kg bodyweight) as the 0·5 per cent eprinomectin solution. The treatments were applied along the topline from the withers to the tailhead. Live lice were counted before treatment on day –1 or day 0, and then at weekly intervals for eight weeks. Six predilection sites per animal were selected for lice counts. Sample sites included the face, neck, withers, topline (midway between the withers and tailhead), tailhead and rump. In the absence of lice at those sites, a thorough body search was undertaken. All live lice present at each site were counted in studies 1 and 3, and in study 2 counts above 100 were recorded as ‘>100’. If the maximum of 100 lice was reached at one site, the animal’s total count and any mean values calculated with that count were preceded by ‘>’. Lice counts for all sites on an individual animal for a given day were added together to obtain that animal’s total lice count. Individual total lice counts were transformed to the natural logarithm (count + 1) for the calculation of geometric group means. Treatment comparisons were made at each point after treatment using Wilcoxon’s rank sum statistic. The results for the efficacy of eprinomectin against L vituli and B bovis are given in Table 2. Lice were eliminated in two of the studies or greatly reduced in one by day 7, and two weeks after treatment no living B bovis or L vituli were found on any eprinomectin-treated calves; the louse infestation of the controls was maintained throughout the studies. The mean L vituli counts for the controls remained constant or increased during the study period, whereas the B bovis counts decreased. The difference between the louse counts for the treated and control calves was statistically significant (P<0·01) for each investigation day after treatment. Veterinary Record (2005) 156, 112-113

Study of the sustained speed of kill of the combination of fipronil/amitraz/(S)-methoprene and the combination of imidacloprid/permethrin against Dermacentor reticulatus, the European dog tick

The sustained speed of kill against Dermacentor reticulatus of two topical combinations, one containing fipronil/amitraz/(S)-methoprene and the other, imidacloprid/permethrin, was evaluated in dogs. Two treated groups and one untreated control group of eight adult Beagle dogs each were randomly formed based on pre-infestation rates and bodyweight. Each treatment was administered topically once on Day 0, according to the recommended label dose and instructions for use. All dogs were infested with 50 adult unfed D. reticulatus starting on Day 1, then weekly, for a total of five weeks. While most studies determine tick efficacy at 48 hours (h), in this study, all remaining ticks were counted and categorized 24 h following each infestation. The numbers of ticks (living or dead) that remained attached on treated dogs were compared to those on the control animals. The percent reduction of attached ticks (disruption of attachment) at 24 h on dogs treated with fipronil/amitraz/(S)-methoprene remained above 92% for four weeks. The reduction of attached ticks at 24 h on dogs treated with imidacloprid/permethrin did not reach 80% during the entire study. The number of ticks attached at 24 h was significantly (p < 0.05) lower in the fipronil/ amitraz/(S)-methoprene group than in the imidacloprid/permethrin group in assessments on Days 2, 15, 22, 29 and 36. When assessing efficacy based upon live ticks on treated versus control dogs, fipronil/amitraz/(S)-methoprene 24 h efficacy was above 95% for four weeks, decreasing to 77.8% at Day 36. The 24 h efficacy of imidacloprid/permethrin ranged from 56.2% to 86.7% through Day 29, never achieving 90% throughout the study. The 24-hour efficacy of fipronil/amitraz/(S)-methoprene was significantly (p < 0.05) higher than imidacloprid/permethrin at all time points, including Day 36.

Efficacy of a topical application of Certifect® (fipronil 6.26% w/v, amitraz 7.48% w/v, (S)-methoprene 5.63% w/v) for the treatment of canine generalized demodicosis

The efficacy of the treatment with Certifect ® (containing fipronil 6.26% w/v, amitraz 7.48% w/v, (S)-methoprene 5.63% w/v) applied topically was assessed in 18 dogs diagnosed with clinical generalized demodicosis. Three treatment regimens were compared over a 3-month period. Starting at Day 0, dogs were treated monthly (group 1) or every two weeks (group 2) with the combination of fipronil, amitraz, and (S)-methoprene or with monthly topical applications of the combination of amitraz and metaflumizone (group 3, reference treatment). Clinical examinations including deep skin scrapings were performed every month in order to evaluate the resolution of clinical signs and the reduction in mite counts. On Day 84, the percentage reduction of mite counts in group 1 was 99.8%, whereas no Demodex canis could be detected in groups 2 and 3 (i.e. 100% parasitological efficacy). As a result of the Demodex mite count reduction, the skin condition of the dogs improved significantly in all groups. This study illustrates, that both monthly and bi-weekly treatments with Certifect were effective in treating dogs with generalized demodicosis over a 3-month period.

Efficacy of a single dose of a novel topical combination product containing eprinomectin to prevent heartworm infection in cats.

Cats may be infected by heartworm, Dirofilaria immitis, through mosquito bites. They can develop severe heartworm disease when infective D. immitis larvae migrate and develop into adults in the pulmonary vasculature or other tissues. As there is no curative treatment for feline heartworm infection, the monthly administration of preventative treatment is recommended in endemic areas. Three controlled, blinded laboratory studies were conducted to evaluate the preventative efficacy of BROADLINE(®), a novel combination of fipronil, (S)-methoprene, eprinomectin, and praziquantel against D. immitis in cats. In each study, 28 cats were inoculated with approximately 100 (studies 1 and 2) or 40 (study 3) infective third stage D. immitis larvae by subcutaneous injection, thirty days prior to treatment. The larvae were from recent field isolates from naturally infected dogs from three distinct geographic areas (two in the USA and one in Europe). In each study, the cats were allocated randomly to two study groups of 14 cats each. The control group remained untreated. On Day 0, each cat in the treated group received one topical application of the novel topical formulation, delivering the minimum intended dose of 0.5mg of eprinomectin per kilogram of body weight. At 6 months after infection, all cats were humanely euthanized and examined for adult D. immitis. Across all three studies, 28 (68%) of the 41 untreated cats harbored one or more heartworms, while 100% of the 42 treated cats remained free of heartworm infection, demonstrating the 100% preventive efficacy of BROADLINE(®) against D. immitis in cats. The treatment was well tolerated and no health abnormality was observed in any treated cat.