Topical Combination Of Fipronil Research Articles

Repellent efficacy of a new combination of fipronil and permethrin against Lutzomyia longipalpis

BackgroundLutzomyia longipalpis is the main vector of Leishmania infantum, the agent of canine and human visceral leishmaniosis in the Americas. Considering that the dog is the main domestic host of the parasite, repellent treatment is a measure that might contribute to the prevention of canine visceral leishmaniosis. The repellent efficacy of a single treatment of a new spot-on topical combination of fipronil and permethrin (Frontline Tri-Act®, Merial, now part of Boehringer-Ingelheim) to repel Lu. longipalpis sand flies was evaluated.MethodsSixteen healthy Beagle dogs, eight females and eight males, weighing 8.4–14.4 kg, and 2 to 4 years-old were included in the study. Animals were blocked on decreasing body weight and randomly allocated within the blocks to one of two treatment groups of eight animals each. Dogs in Group 1 were untreated and Dogs in Group 2 were treated with a combination containing 67.6 mg/ml fipronil + 504.8 mg/ml permethrin (Frontline Tri-Act®) once on Day 0. Sand fly exposures were performed on Days 1, 14, 21 and 30 with Lu. longipalpis female sand flies. After 65 (± 15 min), sand flies were assessed for engorgement status.ResultsThe percent repellency of the treated group compared to the untreated control group was 95.7, 94.3, 81.7 and 72.2% for exposure days 1, 14, 21 and 30, respectively. The two treatment groups were significantly different for all exposure days (P ≤ 0.016 for days 1, 14, 21 and 30). No adverse reactions were observed during the study.ConclusionA single topical administration of a new combination of fipronil and permethrin demonstrated a significant repellent effect against Lu. longipalpis bites as soon as it was applied on the dogs and its repellent efficacy lasted for 4 weeks with results greater than 80% for 3 weeks. The results suggest that in endemic areas the regular application of the new combination could contribute to protect dogs from Leishmania infection and therefore serve as an additional tool for the prevention of canine visceral leishmaniosis.

Knock-down and speed of kill of a combination of fipronil and permethrin for the prevention of Ctenocephalides felis flea infestation in dogs.

BackgroundA topical combination of fipronil + permethrin (Frontline Tri-Act®/Frontect®, Merial) has recently been developed to control fleas, ticks, mosquitoes, sandflies and stable flies on dogs. Two studies were conducted to assess its speed of kill and knock-down effect on Ctenocephalides felis fleas. The combination was compared to either fipronil alone or to a combination of permethrin, dinotefuran, and pyriproxyfen,MethodsIn each study, 18 dogs were randomly allocated to one of three groups: (Group 1: untreated dog; Group 2: treated once on D0 with the combination of fipronil and permethrin; Group 3: treated once on D0 either with fipronil alone (study 1) or with a combination of permethrin, dinetofuran and pyriproxyfen (study 2)). Each dog was infested with 100 unfed adult C. felis fleas on Days 2 (study 2), 7, 14, 21 and 28. Fleas were collected from dogs at 1 h and 12 h post- infestations (PI) (study 1) or at 2 h and 6 h PI (study 2) to assess efficacy and from collection pans underneath cages 1 h (study 1) or 5 min (study 2) PI to assess knock-down effect.ResultsAll treated dogs had significantly (p ≤ 0.01) lower flea counts than untreated dogs at every time point in both studies. For a whole month, a significant knock-down effect against infesting fleas is obtained in five minutes PI with the combination of permethrin and fipronil. Complete efficacy (>95 %) was achieved in 1 h (study 1) or 2 h (study 2) PI for 14 days and by 6 h PI for all challenges conducted throughout the month. Efficacy remains >85 % at 2 h PI for the whole month. A significantly higher efficacy of the fipronil + permethrin combination compared to other treatments was demonstrated at the earliest time points for the month (1 h knock-down effect and insecticidal efficacy compared to fipronil alone; 5 min knock-down effect compared to the combination of permethrin + dinetofuran + pyriproxyfen).ConclusionsThe rapid flea knock-down effect and speed of kill demonstrated by the spot on combination of fipronil + permethrin provide a reliable strategy against flea infestations in dogs.

Efficacy of a Novel Topical Combination of Fipronil 9.8% and (S)-Methoprene 8.8% against Ticks and Fleas in Naturally Infested Dogs.

The efficacy of a novel topical combination of fipronil 9.8% (w/v) and (S)-methoprene 8.8% (w/v) (Fiprofort® Plus) was tested against ticks and fleas in naturally infested dogs. A total of fifty dogs were allocated in the study with ticks infestation (n = 35) and fleas infestation (n = 15). On day 0, thirty-five tick and fifteen flea infested dogs received the test formulation, a combination of fipronil 9.8% (w/v) and (S)-methoprene 8.8% (w/v) spot-on solution. Ticks and flea counts were taken on days 0 (pretreatment) and 3, 7, 14, 21, 28, and 35 after treatment. Blood samples were collected for evaluation of haematological parameters on days 0 (pretreatment) and 7, 21, and 35 after treatment. All the adult ticks and fleas collected were identified as Rhipicephalus sanguineus and Ctenocephalides felis, respectively. The efficacy of spot-on formulation against ticks was 34.00% (day 3), 53.14% (day 7), 62.71% (day 14), 65.48% (day 21), 59.80% (day 28), and 58.82% (day 35), whereas against fleas it was 38.00% (day 3), 64.34% (day 7), 89.67% (day 14), 95.40% (day 21), 100.00% (day 28), and 100.00% (day 35). Haematological parameters for ticks and fleas infested dogs were statistically nonsignificant as compared to control. The combination of fipronil and (S)-methoprene eliminated the existing ticks and fleas infestation and prevented the dogs from flea and tick infestation for four weeks.

Efficacy of Broadline® spot-on against Aelurostrongylus abstrusus and Troglostrongylus brevior lungworms in naturally infected cats from Italy

The increasing reports of Aelurostrongylus abstrusus infection and the new information on Troglostrongylus brevior have spurred the interest of the scientific community towards the research of pharmaceutical compounds effective against both pathogens. A novel topical combination of fipronil, (S)-methoprene, eprinomectin and praziquantel (Broadline®, Merial) has been released for the treatment of a variety of feline parasitic infections. The present study reports the efficacy of this spot-on in treating cats naturally infected by feline lungworms. Client owned cats (n=191) were enrolled from three geographical areas of Italy and faecal samples were examined by floatation and Baermann techniques. Twenty-three individuals were positive for L1 of A. abstrusus (n=18) or T. brevior (n=3) or for both species (n=2) and they were topically treated with Broadline®. Seventeen of them were also concomitantly infected by other parasites. Four weeks after treatment, faecal samples were collected and examined to assess the efficacy of a single administration of the product. Based on lungworm larvae counts, the efficacy of the treatment was 90.5% or 100% for A. abstrusus or T. brevior, respectively. Cats released significantly lower amounts of lungworm larvae after treatment compared to pre-treatment (p<0.0001). All but three cats were negative for other nematodes after treatment and all cats recovered from respiratory signs. Results of this study indicate that a single administration of the topical combination fipronil, (S)-methoprene, eprinomectin and praziquantel is effective and safe for the treatment of A. abstrusus and/or T. brevior infections in cats living under field conditions.

Sustained speed of kill and repellency of a novel combination of fipronil and permethrin against Ctenocephalides canis flea infestations in dogs.

BackgroundCtenocephalides canis is a major flea species in dogs in several European countries. The new topical combination of fipronil and permethrin (Frontline Tri-Act®/Frontect®, Merial) has been developed to control fleas, ticks, mosquitoes, sandflies and biting flies on dogs. Considering the repellent and insecticidal effects of permethrin and the insecticidal effect of fipronil, the efficacy of the combination against fleas including C. canis was expected to be rapid. The study was conducted to measure the 1-hour, 6-hour and 24-hour efficacy, as well as the repellent activity, of the fipronil-permethrin combination on treated versus untreated dogs.Methods12 Beagle dogs were randomly allocated to one of two groups based on pre-treatment live flea counts. Dogs in Group 1 remained untreated whereas dogs in Group 2 were treated once on Day 0. Each dog was infested with 100 unfed adult C. canis on Days 2, 7, 14, 21 and 28. Dogs were combed for fleas 1 and 6 h after each infestation. Following this examination, fleas remaining on the liner at the bottom of each cage were collected and counted. All live fleas were placed back on each dog after the 1- and 6-hour counts. A comb-count was performed at 24 h post infestation on all dogs.ResultsTreated dogs had significantly (p ≤ 0.01) lower flea counts than untreated dogs at every time point. The percent efficacy was ≥99.1% at 6 and 24 h after each weekly challenge up to the month. The 1-hour counts also showed good efficacy of 96.5%, 98.9%, 92.0%, 70.2% and 55.7% on Days 2, 7, 14, 21 and 28, respectively. The repellent efficacy, assessed on the liners at 1 h, was 86.5%, 94.9%, 79.5%, 58.4% and 43.9% on Days 2, 7, 14, 21 and 28, respectively.ConclusionsThis study demonstrates the beneficial effect of the fipronil and permethrin combination against C. canis, providing both a repellent and insecticidal effect as early as 1 h post infestation, and >99.1% efficacy calculated at 6 h during a month.

Therapeutic efficacy of Broadline against notoedric mange in cats.

The efficacy of a novel topical combination of fipronil 8.3% w/v, (S)-methoprene 10 % w/v, eprinomectin 0.4% w/v, and praziquantel 8.3% w/v (Broadline, Merial) was evaluated in 18 cats naturally infested by Notoedres cati in a controlled, blinded clinical efficacy study. Cats were blocked on pretreatment bodyweight and randomly allocated to two groups of nine cats each. One group served as control (untreated) and one group was treated once topically with Broadline according to the label instructions. Skin scrapings (three scrapings per animal per occasion) were collected prior to treatment and every other week for 8 weeks thereafter and examined for live N. cati mites. In addition, lesions were evaluated at each sampling to monitor the clinical recovery. Based on live mite counts, efficacy against N. cati of a single topical administration with Broadline was >99%, and all treated cats recovered from clinical signs of notoedric mange. No treatment-related adverse events were observed.

Efficacy of a novel topical combination of fipronil, (S)-methoprene, eprinomectin and praziquantel against larval and adult stages of the cat lungworm, Aelurostrongylus abstrusus

The efficacy of a novel topical combination of fipronil 8.3% w/v, (S)-methoprene 10% w/v, eprinomectin 0.4% w/v, and praziquantel 8.3% w/v (BROADLINE®,11Broadline® is a trademark of Merial; all other marks are the property of their respective owners. Merial) against larval and adult Aelurostrongylus abstrusus lungworms in cats was assessed in a controlled laboratory study. The study included 48 purpose-bred, short-haired cats which were each inoculated with 225 infective A. abstrusus larvae. The cats were formed into eight blocks based on pre-treatment bodyweight and were then, within each block, randomly allocated to one of six treatment groups: untreated control; treated once when A. abstrusus were expected to be third-stage larvae (4 days post inoculation [dpi]), fourth-stage larvae (7dpi), immature adults (14dpi) or adult nematodes (32dpi), or treated twice, once when A. abstrusus were expected to be third-stage larval and once again when A. abstrusus were expected to be adult nematodes (4dpi+32dpi). Cats weighing ≥0.8–2.5kg received one 0.3mL applicator and cats weighing >2.5–7.5kg received one 0.9mL applicator. For determination of the efficacy of treatments, lungworm larval counts were established on faecal samples collected from all cats 32, 39, 46, 53 and 60dpi. At each occasion from 46dpi on, cats treated with fipronil, (S)-methoprene, eprinomectin and praziquantel had significantly lower A. abstrusus larval counts than the untreated controls with percentage reductions of 91.6% (cats treated 14dpi; P=0.012), ≥98.9% (cats treated either 4dpi, 7dpi or 32dpi; P<0.001) or >99.9% (cats treated 4dpi+32dpi; P<0.001) at 60dpi. Thus, the novel topical combination of fipronil, (S)-methoprene, eprinomectin and praziquantel was highly effective in the prevention and treatment of A. abstrusus lungworm infection in cats.

Efficacy of a novel topical combination of fipronil, (S)-methoprene, eprinomectin and praziquantel against adult and immature stages of the cat flea (Ctenocephalides felis) on cats

The efficacy of a novel topical combination of fipronil 8.3% (w/v), (S)-methoprene 10% (w/v), eprinomectin 0.4% (w/v) and praziquantel 8.3% (w/v) (BROADLINE®) was tested against adult and immature stages of Ctenocephalides felis fleas in six studies. For that purpose, fleas from different colonies from North America, Germany and South Africa were used to induce infestations in cats under laboratory conditions. In each study, between 12 and 16 cats were allocated randomly to 2 groups. Cats in Group 1 were not treated and served as controls. Cats in Group 2 were treated once on Day 0 with BROADLINE® at the minimum recommended dosage of 0.12mg/kg body weight. In 4 studies, all animals were infested experimentally with unfed C. felis (100±5) on Days 2 (or 1), 7, 14, 21, 28 and 35. Live fleas were counted 24h post-treatment or infestation. In 2 additional studies, animals were infested at the same frequency with gravid C. felis fleas (100±5) that were fed previously on an untreated host. Forty-eight hours post-infestation, flea eggs were collected, counted and incubated for the evaluation of the reduction of emergence of adults. The combined curative efficacy against adult fleas at 24h after treatment was 94.3% and the combined preventive efficacy values remained greater than 95.9% at 24h after 5 subsequent weekly infestations. In addition, the product reduced dramatically the emergence of new adult fleas for at least 5 weeks (>98.1% for one month and 93.2% at 5 weeks after infestation), demonstrating its efficiency in preventing environmental contamination by immature stages.

Efficacy of a novel topical combination of fipronil, (S)-methoprene, eprinomectin and praziquantel against induced infections of Ancylostoma spp. nematodes of cats

Four studies were conducted to examine the efficacy of a novel topical combination of fipronil, (S)-methoprene, eprinomectin, and praziquantel (BROADLINE®, Merial) against Ancylostoma tubaeforme and Ancylostoma braziliense hookworms of cats. In each study, purpose-bred cats were randomly assigned to treatment groups of 10 or 12 cats per group. In three studies the cats were inoculated with A. tubaeforme and in one study with A. braziliense. The inoculations were undertaken on a schedule which resulted in the hookworms reaching the fourth larval stage in two of the studies, or the adult stage in four of the studies, by the day of treatment. In each study there was also an untreated control and 1 or 2 groups treated with the novel combination. In the two studies where efficacy against the fourth larval stage of A. tubaeforme was tested, the efficacy recorded was 100%. In the three studies where efficacy against the adult stage of A. tubaeforme was tested, efficacy of 100% was also confirmed. In the study where efficacy against the adult stage of A. braziliense was tested efficacy was 99.5%.

Efficacy of a novel topical combination of fipronil, (S)-methoprene, eprinomectin and praziquantel against adult and larval stages of Toxocara cati in cats

The efficacy of a novel topical combination of fipronil 8.3% (w/v), (S)-methoprene 10% (w/v), eprinomectin 0.4% (w/v), and praziquantel 8.3% (w/v) (BROADLINE®, Merial) was evaluated against adult and larval Toxocara cati in four controlled studies. All studies included experimentally infected, purpose-bred, short-haired cats. In two studies, 22 or 20 cats harbouring patent infections as confirmed by pre-treatment faecal examination, were included. Within each study, cats were allocated to one of two groups: control or treated. In a further two studies, 30 cats were included in each; cats were allocated to one of three groups: control, treated when T. cati were expected to be either migrating third and/or fourth-stage larvae, or treated when T. cati were expected to be fourth-stage larvae. Cats allocated to the treated groups received a single topical application of the combination product at 0.12mL/kg bodyweight (10mg fipronil+12mg (S)-methoprene+0.5mg eprinomectin+10mg praziquantel per kg). For parasite recovery and count, cats were euthanized humanely at different intervals after treatment. In the studies targeting adult T. cati, ascarids were recovered from all controls (range 1–150) while only two worms were isolated from one treated cat. Thus, the efficacy of the novel combination was 99.4% and 100% against adult T. cati. For studies targeting larval T. cati, up to 21 worms were recovered from each of seven or eight of the control cats per study. No T. cati were recovered from the treated cats in two studies, corresponding to 100% efficacy against both, migrating third and/or fourth-stage larvae and luminal fourth-stage larvae. All cats accepted the treatment well and no adverse experiences or other health problems were observed throughout the studies.

Efficacy in cats of a novel topical combination of fipronil, (S)-methoprene, eprinomectin, praziquantel, against induced infestations of Echinococcus multilocularis

Although foxes are the main reservoir of Echinococcus multilocularis, it is recognized that dogs and cats also may become infected. In cats the infection and egg production rates are usually low. Nevertheless, cats are a potential source of transmission of E. multilocularis. Due to the high human medical significance of E. multilocularis infection, it is important in endemic areas that owned cats are dewormed regularly. This paper presents the efficacy results of a new topical formulation, Broadline® (Merial) tested against E. multilocularis infection in cats. Two blinded laboratory studies were conducted to evaluate this novel topical combination of fipronil, (S)-methoprene, eprinomectin, and praziquantel against E. multilocularis. In each study, purpose-bred cats were assigned randomly to two treatment groups of 10 cats each: one untreated control group and one group treated at the minimum therapeutic dose of 0.12mL/kg bodyweight to deliver 10mg fipronil, 12mg (S)-methoprene, 0.5mg eprinomectin and 10mg praziquantel/kg bodyweight. The cats were inoculated orally with E. multilocularis protoscolices, 22 or 23 days before treatment. Based on necropsy and intestinal worm count, 8 or 11 days after treatment, the two studies confirmed 100% efficacy of Broadline® against adult E. multilocularis.

Efficacy of a novel topical combination of fipronil, (S)-methoprene, eprinomectin and praziquantel against feline urinary bladder worm (Capillaria plica) infection

Infection with urinary capillarid bladder worms has been observed in cats worldwide. Although considered as generally causing no or little harm, infection with urinary capillarids may be associated with clinical disease which requires an appropriate treatment including the use of anthelmintics. Therefore, the efficacy of a novel topical combination formulation of fipronil 8.3% (w/v), (S)-methoprene 10% (w/v), eprinomectin 0.4% (w/v), and praziquantel 8.3% (w/v) (BROADLINE®, Merial) was evaluated against urinary capillarids in naturally infected cats. Sixteen European Short Hair cats (5 male, 11 female) with capillarid eggs in their urine pre-treatment were included in the study. At the time of treatment, the cats were approximately ten months to eight years old and weighed 1.6–3.6kg. Cats were ranked based on decreasing bodyweight and then randomly allocated within replicates of two animals to one of the treatment groups. Each cat in the treated group received one topical application of the combination product at the minimum therapeutic dose of 0.12mL/kg body weight delivering 10mg fipronil+12mg (S)-methoprene+0.5mg eprinomectin+10mg praziquantel per kilogram of body weight while the cats allocated to the control group remained untreated. For parasite recovery, identification and count, cats were euthanized humanely 14 days after treatment. All untreated cats harboured Capillaria plica in their urinary bladders (range 4–12), while no capillarids were recovered from the eight treated cats. Thus, the efficacy of the novel topical combination against C. plica was 100%. All cats accepted the treatment well based on post-treatment observations and daily observations thereafter. No adverse events or other health problems were observed during the study.

Efficacy of a novel topical combination of fipronil, (S)-methoprene, eprinomectin and praziquantel against experimental infections of Toxascaris leonina in cats

The efficacy of a novel topical fipronil, (S)-methoprene, eprinomectin and praziquantel combination product (BROADLINE®, Merial) was evaluated against adult Toxascaris leonina ascarids in experimentally infected cats in two controlled studies under an identical protocol. For each study, 30 nematode-naive, purpose-bred European Short Hair cats were inoculated orally with approximately 300 larvated T. leonina eggs. Twenty-two and 24 cats, respectively, that were shown to be positive for Toxascaris eggs by pre-treatment faecal examination were subsequently included in the two studies. In each study, the animals were allocated randomly to an untreated (control) group or to a treatment group. The treatment was a novel topical combination: fipronil (8.3%, w/v), (S)-methoprene (10%, w/v), eprinomectin (0.4% w/v) and praziquantel (8.3% w/v). Treatment was applied on Day 0 at 0.12mL/kg bodyweight. For parasite recovery and count, cats were euthanized humanely seven days after treatment and necropsied. All untreated cats harboured adult T. leonina (range, 1–31 nematodes). The treatment provided a high level of efficacy against adult T. leonina in both studies (95.8% and 98.1%, respectively p<0.001). All cats accepted the treatment well based on hourly post-treatment observations for 4h and daily observations thereafter. No adverse experiences or other health problems were observed throughout the studies. Thus the data indicate that this novel combination product will provide a safe and effective treatment against T. leonina in cats.

Efficacy of a novel topical combination of fipronil, (S)-methoprene, eprinomectin, and praziquantel, against the ticks, Ixodes ricinus and Ixodes scapularis, on cats

Five controlled, blinded and randomized studies were conducted to examine the efficacy of a single topical application of a combination of fipronil, (S)-methoprene, eprinomectin, and praziquantel (BROADLINE®, Merial) against induced infestations with Ixodes ticks on cats. Three studies investigated the efficacy against Ixodes ricinus and two against Ixodes scapularis. In each study, purpose-bred cats were assigned at random to an untreated group or to a treated group. For the studies using I. ricinus, cats were infested with 50 female ticks and a similar number of males 2 days before treatment application, and weekly afterwards on between four and six occasions. For the studies using I. scapularis, cats were infested with a total of 50 ticks (approximately 25 females and 25 males) according to the same schedule as for I. ricinus. Tick counts for the evaluation of efficacy were performed 48h after treatment and 48h after the subsequent weekly infestations. Weekly attachment rates to untreated cats of at least 29% for I. ricinus and at least 30% for I. scapularis demonstrated consistently that the ticks were vigorous and that the attachment rates were adequate for efficacy evaluation. In the I. ricinus studies, an efficacy of at least 93% was demonstrated for up to 37 days after the treatment. In the I. scapularis studies, the efficacy level was at least 95% 30 days after the treatment. The product was well tolerated and caused no adverse reaction.

Pharmacokinetics and metabolism of eprinomectin in cats when administered in a novel topical combination of fipronil, (S)-methoprene, eprinomectin and praziquantel

Four studies were conducted to determine the pharmacokinetic characteristics and in vitro metabolism of eprinomectin, a semi-synthetic avermectin, in cats. Pharmacokinetic parameters including bioavailability of eprinomectin were determined in a parallel study design comprised of one group of eight cats which were treated once topically at 0.12mL/kg bodyweight with BROADLINE®, a novel combination product (fipronil 8.3% (w/v), (S)-methoprene 10% (w/v), eprinomectin 0.4% (w/v) and praziquantel 8.3% (w/v)), delivering a dose of 0.5mg eprinomectin per kg body weight, and a group of six cats which received 0.4% (w/v) eprinomectin at 0.4mg/kg bodyweight once by intravenous injection. For cats treated by topical application, the average eprinomectin (B1a component) maximum plasma concentration (Cmax) was 20ng/mL. The maximum concentrations were reached 24h after dosing in the majority of the animals (six of eight cats). The average terminal half-life was 114h due to slow absorption (‘flip-flop’ kinetics). Following intravenous administration the average Cmax was 503ng/mL at 5min post-dose, and the mean elimination half-life was 23h. Eprinomectin was widely distributed with a mean volume of distribution of 2390mL/kg, and the clearance rate was 81mL/h/kg. Mean areas under the plasma concentration versus time curves extrapolated to infinity were 2100ngh/mL and 5160ngh/mL for the topical and intravenous doses, respectively. Topical eprinomectin was absorbed with an average absolute bioavailability of 31%. In a second parallel design study, the dose proportionality of eprinomectin after single topical administration of BROADLINE® was studied. Four groups of eight cats each were treated once topically with 0.5, 1, 2 or 5 times the minimum recommended dose of the combination, 0.12mL/kg bodyweight. Based on comparison of areas under the plasma concentration versus time curves from the time of dosing to the last time point at which eprinomectin B1a was quantified, and Cmax, dose proportionality was established. In addition, the metabolic pathway of eprinomectin using cat liver microsomes, and plasma protein binding using cat, rat, and dog plasma were studied in vitro. Results of the analyses of eprinomectin B1a described here showed that it is metabolically stable and highly protein bound (>99%), and thus likely to be, as with other species, excreted mainly as unchanged parent drug in the feces of cats.

New method using quantitative PCR to follow the tick blood meal and to assess the anti-feeding effect of topical acaricide against Rhipicephalus sanguineus on dogs

A 28-day study was conducted to assess the dynamic of blood feeding by Rhipicephalus sanguineus ticks on dogs treated or not with a novel topical combination of fipronil, amitraz and (S)-methoprene. Dogs were infested weekly through exposure to ticks in crates for 4h. Ticks were then counted in the crates at 2h and 4h post dog exposure. Ticks were also counted and removed from the dogs at 2h, 4h, 6h, 12h and 24h post tick exposure. The inhibition of blood feeding was assessed by both tick quantification and designing and performing a quantitative PCR (qPCR) to detect the canine hydroxymethylbilane synthase (HMBS) gene in ticks. The percentage of repellency sensu lato based on the ticks collected in crates at 2h varied from 4.7% at day 28 to 48.3% at day 7. The immediate mortality rate of the ticks expelled at 2h varied from 1.5% at day 21 to 31.7% at day 7. The efficacy calculation showed that the acaricidal combination started to kill ticks in as little as 2h. The average efficacy reached 90.0% at 12h post crate challenges and 100% at 24h post exposure in crates. The inclusion of an internal amplification control was used to ensure that no significant template-derived PCR inhibition (≤ 6.2%) affected the overall results. The reduction of blood feeding was significant at 4h (>80.0%) and >99.0% at 24h post tick exposure in the crate. The high repellency rate and the lethal efficacy of CERTIFECT(®) resulted in significantly fewer live attached ticks, consequently reducing blood intake and fluid exchanges.

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 6 h and continued at 12, 18 and 24 h 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 12 h 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.

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 6 h 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.