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A fixed-combination chewable tablet incorporating afoxolaner plus milbemycin oxime (NexGard Spectra ® , Merial) was tested in purpose-bred Beagle dogs for efficacy against adult Ancylostoma ceylanicum hookworms. Sixteen dogs were inoculated each by oral administration of approximately 500 infective larvae of A. ceylanicum . Seventeen days after inoculation, the dogs were weighed and allocated randomly to be treated with afoxolaner plus milbemycin oxime chewable tablets or to remain untreated. Commercial chewable tablets of different strength were combined to deliver doses as close as possible to the minimum effective dose of 2.5 mg afoxolaner plus 0.5 mg milbemycin oxime per kg body weight. Parasites were recovered and counted for determination of efficacy seven days after treatment. All eight dogs that had been left untreated were harboring adult A. ceylanicum (geometric mean, 317.8; range, 210–428) while only one and nine A. ceylanicum were recovered from two of the eight dogs treated with afoxolaner plus milbemycin oxime chewable tablets (geometric mean, 0.5; p < 0.0001). Thus, 99.9% efficacy against induced infection of A. ceylanicum was obtained by the use of oral NexGard Spectra ® at the minimum effective dose. Treatment with afoxolaner plus milbemycin oxime chewable tablets was well accepted and safe. [ABSTRACT FROM AUTHOR]

The objective of this experimental study was to assess the insecticidal efficacy of afoxolaner (NexGard®) against bedbugs (Cimex lectularius) on dogs. For each challenge, 20 bedbugs were placed in two chambers positioned in contact to the dog’s skin for 15 min, after which live fed parasites were counted and incubated for survival evaluations. On Day 0, 7 dogs assigned to the treated group were administered afoxolaner orally at the registered dose. All 14 dogs were challenged on Days 1, 7, 14, 21 and 28, and the collected live fed C. lectularius incubated for 72 h (Day 1), and 72 h and 96 h (Days 7, 14, 21 and 28) for survival evaluation. The percent feeding in the control group ranged from 95% to 98.6% and the percent of live fed bedbugs at 96 h ranged from 99.3% to 100% in the control group, demonstrating the viability of the strain and their capacity to feed on dogs. Significantly fewer live fed bedbugs were counted in the treated group, compared to the control group, at all time-points. The reduction of live fed C. lectularius in the afoxolaner group was 41.4% at 72 h after the Day 1 challenge, and 77.2%, 82.7%, 85.0% and 63.5% at 96 h after the Days 7, 14, 21 and 28 challenges, respectively. It is hypothesized that monthly treatment of dogs with afoxolaner could help in preventing a bed bug population from installing in a household if bedbugs bite dogs in the presence of humans.

In the past decade, canine thelaziosis due to Thelazia callipaeda has been diagnosed in an increasing number of European countries, with endemic areas being identified. A multi-center field trial was conducted in endemic areas in France and Spain to evaluate the efficacy of monthly administrations of the oral milbemycin oxime/afoxolaner combination (NexGard Spectra®) for the prevention of T. callipaeda infection in at-risk dogs. A total of 79 dogs negative for T. callipaeda and with a clinical history of eyeworm infection in the past two years completed the study. Dogs were randomly allocated either to a negative control group (42 dogs) or to the NexGard Spectra® treated group (37 dogs). All dogs were followed up for a 6-month period and assessed monthly for the presence of nematodes on the eyes and for the signs of ocular thelaziosis (e.g., conjunctivitis, keratitis, and ocular discharge). When the presence of nematodes was confirmed, the conjunctival fornix was flushed with a saline solution for parasite recovery and counting, and the dogs were treated appropriately. Recovered parasites were stored in 70% alcohol for subsequent morphological identification. During the course of the study, 57.1% (24/42) of the control dogs were diagnosed positive for Thelazia infection, which illustrates a high incidence rate of parasite infection. Conversely, no eyeworm was recovered from any of the 37 dogs that received NexGard Spectra®. All parasites sampled were confirmed to be T. callipaeda. This clinical field study demonstrated that monthly administrations of NexGard Spectra® provided 100% preventive efficacy against canine thelaziosis.

This experimental study aimed to determine the efficacy of Afoxolaner (NexGard®) to prevent Babesia rossi transmission by Haemaphysalis elliptica ticks on dogs. The study included three groups of seven dogs each. Groups 1 and 2 remained untreated, whereas group 3 dogs received NexGard® on Day 0. All dogs were infested by 50 Haemaphysalis elliptica adult ticks: Group 1 on Day 2, Group 2 on Day 28 and Group 3 on Days 2 and 28. The ticks were originally nymphs having fed on B. rossi infected donor dogs. Their infection rate, assessed by PCR, was 12.8% at Day 2 and 6% at Day 28. On Days 0, 7, 14, 21, 28, 35, 42, 49 and 56, and in case of suspicion of babesiosis, blood samples were collected for blood smears, PCR and ELISA. The B. rossi infection rate in the untreated group 1 was 100% (6/6, as one dog was inadvertently treated on Day 15 and removed from statistical analysis). The infection rate was 57.1% (4/7) in group 2, and 0% (0/7) in the afoxolaner treated group 3 at all time-points until the end of the study on Day 56. After tick removal and count 144 h after each infestation, the control groups had an arithmetic mean of ticks of 23.8 (group 1) and 26.8 (group 2). No tick was recovered from any treated dogs. This study demonstrated that NexGard® protected dogs against infection by B. rossi for at least 28 days.

Twelve healthy dogs were included in this laboratory efficacy study. Six dogs were randomly allocated based on body weight to an untreated control group and six to an afoxolaner (NexGard®) treated group. In the treatment group, afoxolaner was administered orally on Day 0 in accordance with label instructions. On Days 1, 14 and 28, each dog was exposed to 60 unfed female and 10 male Phlebotomus perniciosus sandflies for 1 h. At the end of each exposure period, sandflies were counted and assessed for viability and feeding status. There was no statistical difference in mortality (0.0–5.4%), nor in feeding proportion (61.6–78%) between the control and the treated groups at all 1-h post-exposure assessments. After collection, live fed and unfed sandflies were kept for viability assessments at 48 and 72 h post-exposure. In the untreated control group, the average percentages of live, fed, female sandflies after exposure, on Days 1, 14 and 28, ranged from 51% to 74% at 48 h and from 46% to 57% at 72 h, demonstrating model robustness over the 28 days of the study. Significantly fewer live fed sandflies were recorded for the afoxolaner treated group (p

Twelve healthy dogs were studied in this parallel group, blinded, randomised, and negative controlled efficacy study. On Day -1, the 12 dogs included were ranked within sex in descending order of individual pre-treatment (Day -5) fed mosquito counts and randomly allocated by blocks of two dogs to the untreated control group or the afoxolaner-treated group. NexGard® (Merial, now part of Boehringer Ingelheim Animal Health) was administered orally on Day 0 in accordance with the European label instructions. On Days 1, 7, 14, 21 and 28, all dogs were exposed for a duration of 1 hour to 50 ± 5 unfed Aedes aegypti females. After each exposure, mosquitoes were collected after 1 hour and assessed for viability during collection and at 24 ± 2 hours. The arithmetic (and geometric) mean values of live fed mosquito counts at 24 hours after the exposure periods for the negative control group ranged from 33.7 (32.3) to 49.8 (49.7), indicating that this was a vigorous mosquito strain. There was no significant difference between control and treated groups in the number of live and fed mosquitoes at each 1 hour post-exposure collection time. Based on arithmetic and geometric mean values at 24 hours after each exposure, significantly fewer live fed mosquitoes were recorded in the treated group, compared to the negative control group, throughout the study (p

This study was conducted to assess the prevention of egg laying and the inhibition of the emergence of the cat flea (Ctenocephalides felis) resulting from the application of a combination of fipronil and permethrin (Frontline Tri-Act®/Frontect®, Merial) on dogs. Sixteen healthy dogs were included after pre-treatment live flea counts and randomly allocated to two groups. Eight dogs served as untreated controls and 8 dogs were treated on Day 0 and Day 30 with topical application of fipronil/permethrin at the minimum dose of 6.76 mg/kg fipronil and 50.48 mg/kg permethrin. On days −2, 7, 21, 28, 42 and 56, each dog was infested with 100 fleas. Flea eggs were collected from each dog in individual trays from 12 to 36 h after treatment or each flea re-infestation. All fleas were removed by combing and counted 36 h after treatment or infestations. The collected eggs were counted and incubated for 28 days for larval development and adult emergence assessment. The curative efficacy of Frontline Tri-Act®/Frontect® against adult fleas 36 h after treatment was 95.3% and the efficacy remained 100% after subsequent flea infestations for 8 weeks. Compared to the control group, the treatment reduced egg laying by 84.5% within 36 h after first treatment and was 99.9%, 100%, 100%, 100%, 100% on collection days 7, 21, 29, 43 and 57, respectively. Frontline Tri-Act®/Frontect® reduced by 28.7% the emergence of new adult fleas from eggs laid during the 48 h of pre-treatment infestation. The inhibition of adult emergence from incubated flea eggs could not be assessed after flea re-infestation in the treated group as no eggs were collected.

Otodectes cynotis is a commonly occurring surface mite that can be easily transmitted between suitable hosts, including dogs, causing otocariosis. The activity of the systemic insecticide afoxolaner against O. cynotis has been tested once under experimental conditions, showing a high efficacy. The present study aimed to i) assess the efficacy of two consecutive monthly oral administrations of afoxolaner (NexGard®) against O. cynotis in naturally infested dogs under field conditions and ii) evaluate its impact in reducing bacteria or fungal secondary infections. Dogs, positive for O. cynotis (n = 20), were included in the study and allocated in two groups of ten animals each (G1, control group, and G2, treated group). The first group of ear mite-infested dogs was treated with a placebo, while afoxolaner was administered orally to the second group of dogs at Day 0 (D0) and Day 30 (D30), following label instructions. Otoscopic assessments, deep-swab method and swab samples were performed on all dogs (Days 0, 30, 42) to evaluate the presence or absence of live mites and their number throughout the study, as well as to conduct bacterial and fungal assessments. No adverse events likely related were recorded throughout the study. By Day 42 (D42), all dog's ears were flushed to recover ear mites. All treated dogs became negative, as well as two dogs of the control group. The treatment efficacy of afoxolaner was 100 % based on the arithmetic means of the live mite counts. The clinical scores did not change significantly in the control group, whereas they significantly improved in the treated one from D0 to D30 (p-value = 5.47 10 -5 ). No live mites were present in the afoxolaner-treated group at D42 (p-value = 0.00073). In this field study, two oral administrations of afoxolaner at the recommended dose allowed a complete cure of the infestation. Bacterial and Malassezia pachydermatis infections were detected in both groups, although no significant trend was associated to the ear mite treatment., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Background: The present clinical field trial was conducted to assess the efficacy of a broad-spectrum parasiticide spot-on formulation containing eprinomectin (Broadline ® ) against Thelazia callipaeda eyeworm in naturally infected cats., Methods: Fifteen privately owned cats harboring at least one live adult T. callipaeda were included in the study. Cats were randomly allocated to an untreated control group of seven cats or to a Broadline ® -treated group of eight cats. Cats were treated on Day 0; ocular examinations were performed at inclusion and on Days 7 and 14; eyeworms were recovered and counted on Day 14. The primary efficacy assessment was based on group comparison of number of T. callipaeda on Day 14., Results: Seven days after treatment, six of eight treated cats were negative for eyeworm infection per visual examination, and on Day 14 no eyeworms were found in the treated cats while the seven untreated cats were still infected (geometric mean: 1.97). All cats had inflammatory ocular signs at inclusion; on Day 14, five of eight treated cats had recovered while all untreated control cats were still symptomatic. All collected parasites were confirmed to be T. callipaeda by morphology and molecular characterization., Conclusions: A single treatment with Broadline ® provided 100% efficacy against feline thelaziosis and improved related ocular inflammation signs., (© 2021. The Author(s).)

Giardia intestinalis is a flagellated protozoan responsible for giardiosis (also called giardiasis in humans), the most prevalent and widespread parasitic infection in humans and mammals worldwide. The intestinal microbiota is highly diverse and any alteration in its composition may impact on the health of the host. While studies on the mouse model of giardiosis described the role of the gut microbiota in host susceptibility to infection by the parasite, little is known about the gut microbiota during natural infections in dogs and particularly in puppies. In this study, we monitored naturally G. intestinalis -infected puppies for 3 months and quantified cyst excretion every 2 weeks. All puppies remained subclinically infected during the sampling period as confirmed by fecal examination. In parallel, we performed 16S Illumina sequencing of fecal samples from the different time points to assess the impact of G. intestinalis infection on gut microbiota development of the puppies, as well as gut health markers of immunity such as fecal IgA and calprotectin. Sequencing results revealed that the canine fecal microbiota of Giardia -infected puppies becomes more complex and less diverse with increasing age. In addition, significant differences in the structure of the microbiota were observed between puppies with high and low Giardia cyst excretion. Chronic subclinical G. intestinalis infection appears to be associated with some detrimental structural changes in the gut microbiota. G. intestinalis -associated dysbiosis is characterized by an enrichment of facultative anaerobic, mucus-degrading, pro-inflammatory species and opportunistic pathogens, as well as a reduction of Lactobacillus johnsonii at specific time points. Calprotectin levels increased with age, suggesting the establishment of chronic low-grade inflammation in puppies. Further work is needed to demonstrate whether these alterations in the canine gut microbiota could lead to a dysbiosis-related disease, such as irritable bowel syndrome (IBS) or inflammatory bowel disease (IBD).

This study was conducted to assess the acaricidal efficacy of afoxolaner (NexGard®, Boehringer Ingelheim), and fipronil - permethrin (Frontline® Tri-Act, Boehringer Ingelheim) administered once to dogs experimentally infested with Hyalomma marginatum ticks. Twenty-four Beagle dogs were randomly allocated based on a pre-treatment H. marginatum infestation to an untreated control group, a NexGard® or a Frontline® Tri-Act treated groups. Treatments were administered once on Day 0 as per the products' labels. For the efficacy evaluation, dogs were experimentally infested with 30 adult H. marginatum ticks on Days -2, 7, 28 and 36. In-situ counts were performed at 48 h post-treatment on Day 2 and post-infestations on Days 9, 30 and 38. Ticks were removed and counted at 72 h post-treatment on Day 3 and after each tick infestation on Days 10, 31 and 39. The numbers of live ticks counted in the treated groups were significantly different than in the control group at all time-points (p ≤ 0.0006). The efficacy was at least 97% after 48 h, and at least 99% after 72 h for both treatments. In this study both afoxolaner and fipronil/permethrin formulations demonstrated a high efficacy against adult H. marginatum ticks in treated dogs for at least five weeks., (Copyright © 2021 The Authors. Published by Elsevier B.V. All rights reserved.)

Background: Capillaria aerophila and Capillaria boehmi parasitize the respiratory system of wild and domestic carnivores. Capillaria aerophila inhabits the trachea and bronchi of dogs and cats, while C. boehmi affects the nasal cavities and sinuses of dogs. In dogs the infection may be subclinical or characterized by varying respiratory distress., Methods: The present study evaluated the efficacy of an oral formulation containing milbemycin oxime and afoxolaner (NEXGARD SPECTRA ® ) in dogs naturally infected with C. aerophila and/or C. boehmi from three enzootic areas of Italy. Dogs were enrolled pending fecal examination and molecular confirmation of respiratory capillarioses. Dogs were allocated in two groups: Group 1 (G1, 25 dogs), treated with a negative control product with no anthelmintic activity (afoxolaner, NEXGARD ® ), and Group 2 (G2, 26 dogs), treated with NEXGARD SPECTRA ® . At the day of treatment administration (Day 0), all dogs were clinically examined. Dogs were again subjected to clinical and fecal examinations at Days 28 (± 4) and 56 (± 2). The primary criterion for treatment efficacy was the reduction of fecal Capillaria egg counts in G2 compared with G1. The regression of/recovery from baseline clinical signs was considered as a further efficacy criterion., Results: Percentage reduction of fecal Capillaria egg counts in the NEXGARD SPECTRA ® group compared to the control group was > 97% on Day 28 and 100% on Day 56, respectively (p < 0.05 for both time points). Twelve of the 13 dogs in the NEXGARD SPECTRA ® group with respiratory signs prior to treatment were free of clinical signs at the end of the study. Conversely, the six control group dogs with respiratory signs prior to treatment remained symptomatic., Conclusions: Results of the present study showed that NEXGARD SPECTRA® was safe and highly efficacious in the reduction of C. aerophila and C. boehmi eggs after one treatment with a complete reduction of the egg output after the second administration associated with a recovery from respiratory signs.

Canine vector-borne disease transmission can be reduced with regular use of repellent insecticides. The objective of this year-long experimental study was to assess the efficacy of a topical formulation of fipronil/permethrin (Frontline Tri-Act®) in preventing transmission of Leishmania infantum by sandflies. This clinical field trial was conducted in Xanthi (Northern Greece), an area highly endemic for canine leishmaniosis, from April 2018 to March 2019. Forty purpose-bred Beagle dogs, testing negative for L. infantum prior to study initiation, were enrolled in the study, which consisted of three phases: Phase 1 (field exposure phase) took place from Day 0-196. The dogs were randomly allocated to two groups, group 1 (sham-treated topically with sterile water) and group 2 (treated topically with Frontline Tri-Act®). Dogs in both groups were housed in two subunits of an open-air kennel for a period of 7 months, spanning the Leishmania transmission season. All dogs were treated or sham-treated on Days 0, 28, 56, 84, 112, 140 and 168. Clinical examinations, PCR analysis of conjunctival swabs, and serological tests were performed on a monthly basis. Sandflies were collected every 2 weeks, during a 12 -h period overnight using light traps. Each collection was placed in a container and kept refrigerated until speciation and PCR analysis could be performed. In the second phase of the study, from Day 197 to 252, the dogs were moved into a protected environment (insect-proof protected environment phase). CDC light traps were activated every 2 weeks inside and outside the kennels to ensure the vector-free status of the facility. Monthly clinical examinations, including PCR analysis of conjunctival swabs, and serological tests continued. At the end of the phase 2, bone marrow samples were collected on all dogs. Phase 3 (the final post-winter check) took place from Day 253 to 350. Dogs were released and adopted by individual private owners on Day 253. Follow up analyses included blood collection for SNAP tests and conjunctival swaps for PCR analysis on Days 304 and 350. Additionally, bone marrow collections were also performed on Day 350. Presence of sandflies was observed only in the phase 1 exposure period, and 1714 sandflies were collected (1427 females and 287 males). Two species were identified, Phlebotomus perniciosus var . tobbi and Phlebotomus neglectus . Out of the 62 pooled samples of sandflies assessed by PCR, three were considered positive (4.8 %). By the end of the study, 35 % of the Group 1 dogs (7/20) became positive based on PCR (conjunctival swab and bone marrow) and 30 % (6/20) based on SNAP/IFAT and ELISA tests, while all the dogs in the Frontline Tri-Act® treated group 2 remained negative for all tests (G1 vs G2, p = 0.008). All tests identified the same positive animals, and PCR allowed the detection of one additional infected dog. This clinical field trial demonstrated that monthly administration of Frontline Tri-Act® to dogs exposed to Leishsmania infection in a high endemic area provided 100 % preventive efficacy against transmission of L. infantum ., (© 2020 The Authors.)

Background: A multi-centre field trial was conducted to evaluate the efficacy of afoxolaner based chewables (NexGard® or NexGard Spectra®) for the treatment of generalised demodicosis caused by Demodex canis in dogs under field conditions in France, Italy and Poland., Methods: Client-owned dogs, diagnosed positive for Demodex mites by pre-treatment skin scrapings and presenting clinical signs of generalised demodicosis were included. Dogs were orally treated with afoxolaner three times at monthly intervals. Of the 50 dogs enrolled, 48 completed the whole study. Efficacy of the treatments was assessed monthly by Demodex mite counts and physical examination with special regard to the severity and extension of skin lesions., Results: Treatments were well tolerated in all dogs and resulted in a rapid reduction of mites, with all post-treatment mite counts significantly lower than baseline. The number of mites was reduced by 87.6%, 96.5% and 98.1% on Days 28, 56 and 84, respectively. In addition, the skin lesion severity and extent scores as well as the pruritus were all significantly lower at all post-treatment visits compared to the pre-treatment assessment., Conclusions: This clinical field study demonstrated that monthly administrations of afoxolaner in NexGard® or NexGard Spectra®, offered a convenient and reliable solution for the treatment of canine generalised demodicosis.

Afoxolaner (AFX) plus milbemycin oxime (MO) combination chewable tablets (NexGard Spectra®, Merial) were evaluated for safety and efficacy against naturally acquired nematode infections in domestic dogs in a multi-centre, positive control, blinded field study using a randomized block design based on the order of presentation for allocation. In total, 408 dogs confirmed positive for naturally acquired infections of intestinal nematodes by pre-treatment faecal examination were studied in ten countries in Europe (Albania, Austria, Bulgaria, France, Germany, Hungary, Italy, Lithuania, Romania and Slovakia). Pre-treatment faecal examination revealed Toxocara, Toxascaris, hookworm, Trichuris and/or Capillaria nematode infections in 134, 30, 223, 155 and 14 dogs, respectively. Dogs were allocated to one of two treatment groups in a ratio of 1, AFX + MO chewables (≥2.5 mg AFX + ≥0.5 mg MO per kg body weight, according to dose bands; 207 dogs), and 1, MO plus praziquantel (PRZ) chewables (Milbemax®, Novartis; ≥0.5 mg MO + ≥5 mg PRZ per kg body weight, according to the manufacturer's instructions; 201 dogs) and treated once. For evaluation of efficacy based on reduction of faecal nematode egg counts, two faecal samples, one collected prior to treatment and one collected 9 to 21 days after treatment, were examined using modified McMaster techniques. For evaluation of systemic safety, dogs were examined by a veterinarian before treatment administration and at study end, and dog owners observed the health status of their dogs until the end of the study and reported any abnormal observation. For dogs treated with AFX + MO chewables, the efficacy was 99.7, 99.7, 97.2, 99.7 and 99.7 % for Toxocara, Toxascaris, hookworm, Trichuris and Capillaria, respectively; and the efficacy was 99.5, 99.4, 94.3, 99.9 and 98.0 %, respectively, for the MO + PRZ-treated dogs (p ≤ 0.002 for all nematodes and both treatments). For Toxocara, hookworm and Trichuris, non-inferiority analysis demonstrated that the efficacy of AFX + MO chewable tablets was equal to or better than that of MO + PRZ. In spite that both treatments were ≥98 % efficacious against Toxascaris and Capillaria, a hypothesis of non-inferiority for both genera could not be established due to the low number of dogs infected with these parasites. No treatment-related adverse experiences were observed throughout the study. For both treatments, all dogs were given a systemic safety score of 'excellent' apart from one dog in each treatment group which received a score of 'acceptable'. AFX + MO combination chewables were shown to be safe and demonstrated a high level of efficacy when administered once to dogs infected with a broad range of parasitic nematodes under field conditions.

Background: Infection of dogs with the cardiopulmonary nematode Angiostrongylus vasorum may result in severe clinical disease therefore adequate prevention is necessary. A randomized, negative control, blinded study was conducted to evaluate the efficacy in the prevention of canine A. vasorum infection after monthly administrations of NexGard Spectra®, a novel chewable tablet formulation combining the insecticide and acaricide afoxolaner and the anthelmintic milbemycin oxime, in a multiple challenge (trickle infection) model., Methods: Twenty beagle dogs were challenged orally with doses of approximately 32-43 third-stage larvae of A. vasorum once every other week on seven occasions (Study Days -7, 7, 21, 35, 49, 63 and 77). Ten dogs were administered NexGard Spectra® as close as possible to the minimum recommended dose of afoxolaner and milbemycin oxime, i.e. 2.5 mg/kg body weight and 0.5 mg/kg body weight, respectively, four times at monthly intervals (Study Days 0, 28, 56 and 84) while the remaining ten dogs served as untreated controls. For parasite recovery and count, dogs were euthanized humanely and necropsied six to eight days following the last treatment (Study Days 90-92). Beginning six weeks after first inoculation, faeces were collected on a bi-weekly basis and examined for first-stage larvae of A. vasorum., Results: Untreated dogs harboured 39-95 adult A. vasorum (geometric mean, 66.4), while zero to 24 adult A. vasorum were recovered from the treated dogs (geometric mean, 3.4; P < 0.0001). Thus, efficacy of NexGard Spectra® administered at monthly intervals against incoming A. vasorum was 94.9 %. Compared to the untreated controls, larval excretion of the treated dogs was reduced by 99.9 % (P < 0.0001)., Conclusion: Results of this study demonstrate that NexGard Spectra®, when administered at monthly intervals, can effectively prevent canine A. vasorum infection.

Background: Ticks are hematophageous arthropods that transmit a wide spectrum of pathogens to human and animals. The ability of an acaricidal product to kill ticks quickly provides an important added benefit, especially as protecting dogs from tick bites remains the best preventive measure against tick-borne diseases. The speed of kill of afoxolaner in a novel soft chewable formulation (NexGardTM) against induced infestations with Ixodes ricinus adult ticks was evaluated during a full-month negative controlled and blinded study following a single oral administration., Methods: 12 healthy beagle dogs were included and randomly allocated to 2 groups of six dogs each. One Group was a negative control while the other group was treated with an oral formulation of afoxolaner on Day 0. Tick infestations with 40 (±5) female and 10 male adult unfed I. ricinus were performed on Days -1, 7, 14, 21 and 28. To evaluate immediate efficacy, the number of live ticks were thumb counted at 12 and 24 hours post treatment. To evaluate the persistent speed of kill following further infestations, ticks were thumb counted 12 and 24 hours post infestations. Ticks were removed 24 hours post treatment or infestation., Results: Afoxolaner starts to kill the pre-existing tick infestations rapidly with an immediate efficacy of 93.4% and 100% respectively at 12 h and 24 h post treatment. The persistent speed of kill of afoxolaner was significant (p < 0,05), as compared with untreated controls, at 12 hours after infestations at D7 and D21. Efficacy at 12 h was 76.6%, 41.9%, 36.9% and 38.5% at D7, D14, D21 and D28 respectively. Efficacy at 24 h ranged from 91% to 100% for the entire month., Conclusions: This study demonstrated that besides the excellent acaricidal efficacy of afoxolaner after single oral administration, the product has a rapid speed of kill against one of the most important European tick species and controlled the weekly re-infestations for 28 days post treatment.