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Background Canine babesiosis is a tick-borne disease of significant veterinary importance in dogs. It is caused by Babesia canis in Europe, where it is transmitted by Dermacentor reticulatus ticks. Methods A blinded, randomized, good clinical practice (GCP) and negative control experimental study was conducted to verify the efficacy of NexGard Spectra® in reducing the transmission of B. canis by D. reticulatus to dogs. NexGard Spectra® (IVP) is an oral product for dogs combining afoxolaner, an acaricide/insecticide compound from the isoxazoline class, and milbemycin oxime, a nematicide compound from the macrocyclic lactone class. Three groups of eight dogs were used; one group orally treated on day 0 with the IVP at the minimum recommended dose and two untreated control groups. On day 1, dogs from the treated group and from control group 1 were infested with 50 D. reticulatus adult ticks of 50/50 sex ratio infected with B. canis at a 23% infection rate. On day 28, dogs from the treated group and from control group 2 were infested similarly to those on day 1. Ticks were removed 6 days after each infestation. Results Seven to nine days after each infestation, all untreated control dogs displayed clinical signs of canine babesiosis, i.e., lethargy, and/or dark urine, and/or > 39.5 °C rectal temperature. Blood was collected for microscopical blood smear examination, and for polymerase chain reaction (PCR) analysis. The blood smears from all untreated control dogs were positive for Babesia and all the PCR analyses were positive for B. canis. The control dogs were rescue treated. All control dogs were confirmed positive for B. canis by IFA on day 21 (control group 1) and on day 42 (control group 2). None of the IVP-treated dogs expressed any clinical sign of canine babesiosis following each of the two infestations of days 1 and 28 and until day 56. Blood was collected for IFA and PCR analyses from the treated dogs on days 21, 28, 42, and 56, and all results were negative. Conclusions In this study, the antiparasitic treatment prevented the transmission of B. canis to dogs following induced infestations. Graphical Abstract

Amblyomma maculatum , the Gulf Coast tick, is a species of significant veterinary and public health importance, especially because it is a vector of important diseases, such as American canine hepatozoonosis and tidewater spotted fever. Amblyomma maculatum infests a wide range of vertebrates including livestock, dogs, cats, and humans. Two experimental studies were conducted to evaluate the efficacy of afoxolaner formulated in an oral tablet (NexGard ® ) against induced infestations of A. maculatum in dogs. These Good Clinical Practice (GCP) studies used a randomized, negative controlled and masked design. In each study, 10 dogs were allocated to an untreated group and 10 dogs to a treated group, dosed once on Day 0 with a combination of tablets targeting the minimum therapeutic dose (2.5 mg/kg afoxolaner). Dogs were infested with 50 unfed adult A. maculatum on Days −2, 7, 14, 21, 28, and 35 (Study #1), or on Days −1, 14, and 28 (Study #2). Seventy-two (72) hours after treatment and subsequent infestations, ticks were removed and the numbers of live ticks in each group were used for efficacy calculations. At each time-point, all untreated dogs were adequately infested ( i.e. , with more than 12 live ticks), demonstrating a vigorous tick population and an adequate study model. The curative efficacy against established infestations, 72 hours after treatment, was 100% in Study #1 and 99.5% in Study #2. The preventive efficacy, 72 hours after the post-treatment infestations, ranged from 94.6% to 98.9% for five weeks in Study #1, and was ≥98.8% for four weeks in Study #2. Amblyomma maculatum , la tique du Golfe du Mexique, est une espèce d’importance vétérinaire et de santé publique considérable, notamment parce qu’elle est vectrice de maladies graves, telles que l’hépatozoonose canine américaine et la fièvre pourprée des marées. Amblyomma maculatum infeste un large éventail de vertébrés, notamment le bétail, les chiens, les chats et les humains. Deux études expérimentales ont été menées pour évaluer l’efficacité de l’afoxolaner sous forme de comprimé oral (NexGard ® ) contre les infestations induites d’ A. maculatum chez le chien. Ces études, conformes aux Bonnes Pratiques Cliniques (BPC), ont été randomisées, contrôlées par voie négative et en aveugle. Dans chaque étude, 10 chiens ont été répartis en un groupe non traité et 10 chiens en un groupe traité, recevant une dose unique le jour 0 d’une combinaison de comprimés ciblant la dose thérapeutique minimale (2,5 mg/kg d’afoxolaner). Les chiens ont été infestés par 50 A. maculatum adultes à jeun les jours −2, 7, 14, 21, 28 et 35 (étude n° 1), ou les jours −1, 14 et 28 (étude n° 2). Soixante-douze (72) heures après le traitement et les infestations ultérieures, les tiques ont été retirées et le nombre de tiques vivantes dans chaque groupe a été utilisé pour les calculs d’efficacité. À chaque point temporel, tous les chiens non traités étaient adéquatement infestés (c’est-à-dire avec plus de 12 tiques vivantes), démontrant une population de tiques vigoureuse et un modèle d’étude adéquat. L’efficacité curative contre les infestations établies, 72 heures après le traitement, était de 100% (étude n° 1) et de 99,5 % (étude n° 2). L’efficacité préventive, 72 heures après les infestations post-traitement, variait de 94,6 % à 98,9 % pendant cinq semaines dans l’étude n° 1, et était ≥ 98,8 %, pendant quatre semaines dans l’étude n° 2.

Background Babesia canis is a clinically relevant vector-borne pathogen in dogs, and its presence is expanding. The efficacy of Simparica Trio ® (Zoetis) in the prevention of B. canis transmission was evaluated at the minimum recommended label dose of 1.2 mg/kg sarolaner, 24 µg/kg moxidectin and 5 mg/kg pyrantel per kg bodyweight. Methods Twenty-four (24) dogs were randomly allocated to either a placebo-treated group or one of two treatment groups with Simparica Trio. Dogs were infested with B. canis -infected Dermacentor reticulatus ticks 21 or 28 days after treatment administration. Blood samples for antibody and DNA detection were collected from each dog prior to tick infestation until 28 days after infestation. A dog was defined as being B. canis positive if it tested positive by both an indirect immunofluorescence assay (IFA) and PCR at any time during the study. Results No treatment-related adverse reactions were recorded during the study. All placebo-treated animals displayed clinical signs due to babesiosis and tested positive on both IFA and PCR. None of the Simparica Trio-treated animals displayed any clinical symptoms or tested positive, resulting in a 100% efficacy in the prevention of canine babesiosis ( P  < 0.0001). Conclusions A single treatment with Simparica Trio at the minimum recommended label dose of 1.2 mg/kg sarolaner, 24 µg/kg moxidectin and 5 mg/kg pyrantel per kg bodyweight prevents the transmission of B. canis by infected D. reticulatus to dogs for at least 28 days. Graphical Abstract

Background The ear mite, Otodectes cynotis , is pathogenic, highly contagious and a global cause of otitis externa and pruritus in cats and dogs. The present study evaluated the efficacy of a single oral application of lotilaner flavoured chewable tablets for cats (Credelio™; Elanco, Greenfield, IN, USA) in cats experimentally infested with O. cynotis . Methods Sixteen adult cats were experimentally infested with O. cynotis and confirmed to be mite positive by otoscopic examination. Infested cats were randomly assigned to one of two study groups. On day 0, a group of eight cats was treated once with Credelio™ at 7.0–11.8 mg lotilaner/kg body weight (i.e. at the lower end of the recommended dose range [6–24 mg/kg]), while in the control group eight cats were sham-dosed. All cats were dosed in a fed state. Otoscopic examinations for scoring the number of live mites and the amount of debris/cerumen were performed post treatment (p.t) on days 14 and 28. Ear flushing and microscopic viable mite counts were performed on day 28. Results A single oral dose of Credelio™ on day 0 resulted in a 99.6% reduction in O. cynotis geometric mean mite counts in the Credelio™-treated study group on day 28 recovered by ear flushing. Otoscopic live ear mite count scores on days 14 and 28 did not detect any mites in the Credelio™-treated group. In contrast to this 100% improvement, in the control group an improvement of otoscopic live mite scores was recorded in 25% (day 14) and 12.5% (day 28) of the cats, respectively. Additionally, a comparison of the debris scores to baseline data on day − 2 showed an improvement in 75% of the Credelio™-treated cats on day 14 and in 87.5% on day 28 and no or limited improvement (0% on day 14 and 12.5% on day 28) in the control group animals. Conclusions The results of this study demonstrated that a single application of Credelio™ chewable tablets for cats at the lower end of the recommended dose range was highly efficacious (99.6%) in eliminating experimentally induced O. cynotis infestations and greatly improved clinical signs of otocariosis in cats by 28 days p.t. Graphical Abstract

Esafoxolaner is a purified enantiomer of afoxolaner with insecticidal and acaricidal properties. It is combined with eprinomectin and praziquantel in a novel topical endectoparasiticide formulation for cats. The efficacy of this novel formulation was assessed in an experimental study against induced infestation of Rhipicephalus sanguineus ticks. Twenty cats were randomly allocated to either a placebo control group or a treated group in a 1:1 ratio. Infested cats were treated topically once at the minimum recommended dose. The study was designed to assess curative efficacy 48 h after treatment and to test preventive efficacy 48 h after weekly infestations for 2 months. At each weekly infestation, all cats were infested with 25 male and 25 unfed female R. sanguineus ticks. At each tick count, at least 6 in 10 control cats had a retention of 13 (26%) or more live ticks, demonstrating adequate infestation throughout the study. Curative efficacy on existing tick infestation was 90%; preventive efficacy over the following 6 weeks was at least 96%. L'esafoxolaner est un énantiomère purifié d’afoxolaner, aux propriétés insecticides et acaricides. Il est combiné à éprinomectine et praziquantel dans une nouvelle formulation topique endectoparasiticide pour chats. L’efficacité de cette nouvelle formulation a été testée lors d’une étude contre des infestations expérimentales avec des tiques Rhipicephalus sanguineus. Vingt chats ont été répartis au hasard soit dans un groupe témoin placebo soit dans un groupe traité (rapport 1:1). Les chats infestés ont été traités par voie topique une fois à la dose minimale recommandée. L’étude a été conçue pour une évaluation de l’efficacité curative 48 heures après traitement et pour des évaluations d’efficacité préventive 48 heures après chaque infestation hebdomadaire pendant 2 mois. À chaque infestation hebdomadaire, tous les chats étaient infestés par 25 mâles et 25 femelles de R. sanguineus , non nourris. À chaque comptage, au moins 6 chats sur 10 du groupe placebo contrôle étaient infestés avec au moins 13 (26 %) tiques vivantes, ce qui a validé le modèle d’infestation. L’efficacité curative sur tiques présentes avant traitement a été de 90 %, l’efficacité préventive durant les six semaines suivantes a été d’au moins 96 %.

Background The cestode Dipylidium caninum is known to infect dogs via the ingestion of an intermediate flea host, typically Ctenocephalides felis . Simparica Trio ® is an oral combination drug product for dogs effective in the treatment and prevention of fleas, including C. felis . Here, we report two laboratory studies evaluating the efficacy of a single administration of Simparica Trio at the minimum label dosage of 1.2 mg/kg sarolaner, 24 µg/kg moxidectin, and 5 mg/kg pyrantel (as pamoate salt) in preventing D. caninum infection in dogs for 1 month through killing of C. felis . Methods A total of 20 dogs ( n  = 10 per group) proven to be suitable hosts for C. felis were used in each of the two studies. Treatment occurred on day 0, with each dog given either the placebo or Simparica Trio. On days 0 (after treatment), 7, 14, 21, and 30, dogs were infested using 200 (± 5) unfed D. caninum -infected C. felis . Live flea counts were conducted on day 33 (72 ± 2 h after day 30 infestation). All dogs were euthanized on day 58, and each dog was necropsied for the recovery of D. caninum scolexes from the gastrointestinal tract. Results Placebo-treated dogs had adequate flea infestations and cestode infections in both studies. Simparica-Trio-treated dogs were free of fleas on day 33 (100% efficacy) and had significantly lower mean flea counts compared with placebo-treated dogs ( P  ≤ 0.0007). Scolex counts in Simparica-Trio-treated dogs were also significantly decreased compared with placebo-treated dogs in both studies. The efficacy of Simparica Trio against D. caninum based on least squares mean scolex counts was 100% ( P  < 0.0001) in study 1 and 92.1% ( P  = 0.0033) in study 2. Conclusions The efficacy provided by Simparica Trio against C. felis at the minimum dosage of 1.2 mg/kg sarolaner, 24 µg/kg moxidectin, and 5 mg/kg pyrantel (as pamoate salt) prevented D. caninum infection in dogs for 1 month. Graphical Abstract

Background Revolution ® Plus is a topical combination drug product containing selamectin and sarolaner that has been proven effective against the cat flea Ctenocephalides felis , the intermediate host of the cestode Dipylidium caninum . Here, we report two studies evaluating the efficacy of a single administration of Revolution Plus in preventing D. caninum infection in cats for 1 month through killing of the flea intermediate host. Methods Two studies (study 1 and 2) with the same design were conducted. In both studies, 2 treatment groups of ten cats each were enrolled. On Day 0, the cats in group 1 were treated with a placebo, and the cats in group 2 were treated with Revolution Plus at the minimum recommended dose of 6.0 mg/kg selamectin and 1.0 mg/kg sarolaner. After treatment on Day 0, as well as on Days 7, 14, 21, and 30, the cats in both treatment groups were infested with 100 (± 5) unfed, D. caninum -infected fleas. Live flea counts were conducted on Day 33 (72 ± 2 h after Day 30 infestation). All cats were euthanized on Day 58, and necropsies were performed to enumerate D. caninum scolices in the gastrointestinal tract. Results In both study 1 and 2, all placebo-treated cats were infested with two or more D. caninum scolices at necropsy. Significantly lower mean flea counts were recorded for the Revolution Plus-treated cats compared with placebo-treated cats ( P  ≤ 0.0001), and efficacy based on least squares mean flea counts on Day 33 was 100% (in study 1) and 94.3% (in study 2). Scolex counts were also significantly decreased in Revolution Plus-treated cats compared with placebo-treated cats, with a 97.1% efficacy in study 1 and a 99.3% efficacy in study 2. Conclusions One topical administration of Revolution Plus at the minimum dosage of 6.0 mg/kg selamectin and 1.0 mg/kg sarolaner provided high efficacy in the prevention of D. caninum infection through the killing of its vector, C. felis , for an entire month. Graphical Abstract

Background The parthenogenic reproductive ability of Haemaphysalis longicornis , facilitating quick life cycle completion and rapid geographic spread and its pathogen vector potential make infestations a risk to human and canine health. Two 90-day studies were initiated to evaluate the efficacy of a single fluralaner administration for the treatment and prevention of H. longicornis infestations on dogs. Methods Dogs were randomly assigned (10 dogs/group) to either an untreated control group or a group treated once (Day 0) with 13.64% w/w fluralaner chewable tablets (Bravecto ® ) at the minimum label dose rate of 25 mg/kg. Each dog was infested with approximately 50 H. longicornis ticks on Days -9 or -6 and on Days -2, 28, 58 and 88. A different US tick isolate was used in each study. Tick counts were completed on Days -7 or -4, 2, 30, 60 and 90. The primary efficacy criterion was a 90% reduction in arithmetic mean tick counts between the treated and control groups. For between-group comparisons at any assessment, at least six control dogs were required to retain at least 25% of the infestation dose (13 live ticks). Results Pre-study infestations demonstrated susceptibility of all study dogs to challenge with H. longicornis . At each subsequent assessment in both studies, at least seven untreated control dogs retained ≥ 25% of the challenge, demonstrating adequate infestations for each efficacy calculation. On Days 2, 30, 60 and 90 the mean live tick infestation rate (number of ticks recovered from each dog/infesting challenge of each dog) of untreated control dogs ranged from 27.8 to 60.8%. No live ticks, free or attached, were found on any fluralaner-treated dog in either study. Between-group differences were statistically significant ( P  ≤ 0.0002) at each assessment. Conclusion At the minimum recommended label dose rate of 25 mg/kg, fluralaner chewable tablets were 100% effective in eliminating H. longicornis ticks from dogs infested at the time of treatment. Complete efficacy against both US isolates of this tick was maintained through 90 days following a single treatment. Therefore, fluralaner is a treatment of choice for protecting dogs against this invasive tick species. Graphical abstract

Esafoxolaner, a purified enantiomer of afoxolaner with insecticidal and acaricidal properties, is combined with eprinomectin and praziquantel in NexGard ® Combo, a novel topical endectoparasiticide formulation for cats. The efficacy of this novel formulation against adult and immature stages of Ctenocephalides felis fleas was tested in four experimental studies. Two studies were designed to test adulticide efficacy, one to test inhibition of immature stages, and one to test both adulticide efficacy and inhibition of immature stages. In each study, cats were randomly allocated to a placebo control group or to a novel formulation group treated once at the minimum recommended dose. Cats were experimentally infested weekly for one to two months with unfed C. felis originating from North America or Europe. For adulticide efficacy evaluations, live fleas were counted 24 h after treatment and after subsequent weekly infestations. For immature stages, flea eggs were collected and counted weekly for evaluation of egg production inhibition and incubated for larval hatching evaluation. In the three studies testing adult fleas, curative efficacies, 24 h after treatment, were 92.1%, 98.3% and 99.7%; preventive weekly efficacies, 24 h after weekly infestations, remained higher than 95.5% for at least one month. In the two studies testing immature stages, egg production and larval hatching was significantly reduced for at least one month. These studies provide robust evidence of efficacy of the novel formulation against experimental adult flea infestations and for the prevention of environmental contamination by immature flea stages, for at least one month. L’esafoxolaner, un énantiomère purifié de l’afoxolaner aux propriétés insecticides et acaricides, est associé à l’éprinomectine et au praziquantel dans NexGard ® Combo, une nouvelle formulation endectoparasiticide topique pour chats. L’efficacité de cette nouvelle formulation contre les stades adultes et immatures des puces Ctenocephalides felis a été testée dans quatre études expérimentales. Deux études ont été conçues pour tester l’efficacité des adulticides, une pour tester l’inhibition des stades immatures et une pour tester à la fois l’efficacité des adulticides et l’inhibition des stades immatures. Dans chaque étude, les chats ont été répartis au hasard dans un groupe témoin placebo ou dans un groupe de formulation traité une fois par la nouvelle formulation à la dose minimale recommandée. Des chats ont été expérimentalement infestés chaque semaine pendant un à deux mois par des C. felis non nourris provenant d’Amérique du Nord ou d’Europe. Pour les évaluations de l’efficacité des adulticides, les puces vivantes ont été comptées 24 heures après le traitement et après les infestations hebdomadaires suivantes. Pour les stades immatures, les œufs de puces ont été collectés et comptés chaque semaine pour l’évaluation de l’inhibition de la production d’œufs, et incubés pour l’évaluation de l’éclosion des larves. Dans les trois études testant les puces adultes, les efficacités curatives, 24 heures après le traitement, étaient de 92,1 %, 98,3 % et 99,7 %, et les efficacités hebdomadaires préventives, 24 heures après les infestations hebdomadaires, sont restées supérieures à 95,5 % pendant au moins un mois. Dans les deux études testant les stades immatures, la production d’œufs et l’éclosion des larves ont été considérablement réduites pendant au moins un mois. Ces études fournissent des preuves solides de l’efficacité de la nouvelle formulation contre les infestations expérimentales de puces adultes et pour la prévention de la contamination environnementale par les stades de puces immatures, pendant au moins un mois.

The parthenogenic reproductive ability of Haemaphysalislongicornis, facilitating quick life cycle completion and rapid geographic spread and its pathogen vector potential make infestations a risk to human and canine health. Two 90-day studies were initiated to evaluate the efficacy of a single fluralaner administration for the treatment and prevention of H.longicornis infestations on dogs. Dogs were randomly assigned (10 dogs/group) to either an untreated control group or a group treated once (Day 0) with 13.64% w/w fluralaner chewable tablets (Bravecto.sup.[R]) at the minimum label dose rate of 25 mg/kg. Each dog was infested with approximately 50 H.longicornis ticks on Days -9 or -6 and on Days -2, 28, 58 and 88. A different US tick isolate was used in each study. Tick counts were completed on Days -7 or -4, 2, 30, 60 and 90. The primary efficacy criterion was a 90% reduction in arithmetic mean tick counts between the treated and control groups. For between-group comparisons at any assessment, at least six control dogs were required to retain at least 25% of the infestation dose (13 live ticks). Pre-study infestations demonstrated susceptibility of all study dogs to challenge with H.longicornis. At each subsequent assessment in both studies, at least seven untreated control dogs retained [greater than or equal to] 25% of the challenge, demonstrating adequate infestations for each efficacy calculation. On Days 2, 30, 60 and 90 the mean live tick infestation rate (number of ticks recovered from each dog/infesting challenge of each dog) of untreated control dogs ranged from 27.8 to 60.8%. No live ticks, free or attached, were found on any fluralaner-treated dog in either study. Between-group differences were statistically significant (P [less than or equai to] 0.0002) at each assessment. At the minimum recommended label dose rate of 25 mg/kg, fluralaner chewable tablets were 100% effective in eliminating H.longicornis ticks from dogs infested at the time of treatment. Complete efficacy against both US isolates of this tick was maintained through 90 days following a single treatment. Therefore, fluralaner is a treatment of choice for protecting dogs against this invasive tick species.