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Background Borrelia burgdorferi , the causative agent of Lyme disease, is a zoonotic vector-borne pathogen transmitted by various Ixodes tick species. Lyme disease, while commonly asymptomatic, can induce fever and intermittent lameness in dogs. Highly effective acaricidal products with a rapid onset of action along with prompt removal of attached ticks are important aspects of successful Lyme disease prevention strategies. Methods Two studies were conducted with a total of 30 dogs each. Dogs were randomized to receive a control sham dose, Credelio Quattro, or Credelio. Treatment was administered on Day 0 in a fed state. On Day 28, all dogs were experimentally infested with wild-caught adult Ixodes scapularis . Blood samples for B. burgdorferi antibody analysis utilizing the SNAP 4Dx Plus and Lyme Quant C 6 tests were collected on Days 27, 49, 63, 77, 91, and 105. Skin biopsies were collected from four different areas of heavy tick attachment from each dog for polymerase chain reaction (PCR) detection of B. burgdorferi on Day 104 or 105. Results All control dogs demonstrated adequate I. scapularis infestation rates on Day 33 in both studies. In Study 2, on Day 27, one control dog tested positive for B. burgdorferi on the Lyme Quant C 6 test, prior to experimental tick infestation, and therefore was excluded from analysis. A total of eight out of 10 (Study 1) and nine out of nine (Study 2) control dogs tested positive for B. burgdorferi on at least one test after Day 27. One dog in the Credelio Quattro-treated group tested positive for B. burgdorferi on SNAP 4Dx Plus on Day 105 in Study 1 but was negative on all other tests and study days. None of the dogs treated with Credelio tested positive for B. burgdorferi at any point during either study. Conclusions The laboratory studies described herein confirm that a single dose of lotilaner, at the minimum effective dosage of 20 mg/kg, administered as Credelio Quattro, in combination with moxidectin, praziquantel, and pyrantel or Credelio, is effective for the prevention of transmission of B. burgdorferi from infected I. scapularis for a full month in dogs. Graphical Abstract

Background The invasive tick species, Haemaphysalis longicornis , is becoming established in the USA, presenting a growing threat to dogs and cats. Two 90-day studies were initiated, the same protocol in each, to confirm the efficacy of a single application of two fluralaner formulations against H. longicornis infestations of cats. Methods Cats were randomized among three groups in a 1:1:1 ratio (10 cats/group). Group 1 cats were untreated controls; Group 2 cats were treated with a topical fluralaner formulation (Bravecto®); Group 3 cats received a topical formulation containing fluralaner and moxidectin (Bravecto® Plus). Treatments were administered once (Day 0) at the label dose rates. Each cat was infested with 50 H. longicornis ticks on Day −7 for study qualification and also infested with 50 ticks on Days −2, 28, 58 and 88. Tick counts were completed on Days −5, 2, 30, 60 and 90. The primary objective was based on percentage reductions in arithmetic mean tick counts. Results Pre-study infestations showed all study cats were susceptible to tick challenge. Except for Day 2 in one study, at least six control cats retained ≥ 25% of each challenge, demonstrating an adequate infestation for efficacy assessments. Across studies on Days 2, 30, 60 and 90, the mean live tick infestation rate (number of ticks recovered from each cat/infesting challenge to each cat) of Group 1 cats ranged from 25.0 to 69.6%. Efficacy of each formulation, based on live tick counts, was 100% on Day 2 and > 95 to 100% at each subsequent assessment. Between-group differences were statistically significant ( P  < 0.0001) for each treatment versus control comparison. Conclusion At the label dose rate, both topical fluralaner formulations were 100% effective in eliminating H. longicornis ticks from cats infested at the time of treatment. Efficacy of > 95 to 100% was then maintained through 90 days following a single application. Fluralaner is therefore a treatment of choice for protecting cats against this invasive tick species. Graphical Abstract

Background Haemaphysalis longicornis , the longhorned tick, is an invasive tick species that has been identified in increasing numbers and regions across the USA. This tick species is a competent vector for various pathogens to dogs, humans, and other species, with heavy infestations documented to lead to exsanguination. Therefore, determination of ectoparasiticides providing adequate treatment and control of H. longicornis is imperative to help reduce vector-borne disease transmission and protect against infestation in dogs. Methods Three laboratory studies were conducted to evaluate the efficacy of Credelio Quattro and Credelio for the treatment and control of H. longicornis. A total of 30 dogs per study were randomized to receive either placebo, Credelio Quattro, or Credelio on Day 0 according to a complete block design on the basis of pre-treatment live attached H. longicornis counts, infested on Day −7. To assess efficacy against preexisting infestations, enrolled dogs were infested with 50 unfed adult H. longicornis on Day −2 or −1 prior to treatment. Residual efficacy post-treatment was evaluated with subsequent infestations on Days 5, 12, 19, and 30. All ticks were collected and evaluated as live or dead and free or attached 48 h after treatment or infestation. Results Adequacy of infestation was achieved in at least two studies for every infestation time point evaluated. Both Credelio Quattro and Credelio provided 100% efficacy against H. longicornis from Day 2 through Day 32, with no live ticks observed on any dogs. A statistically significant number of dead ticks were recovered from both treated groups as compared with control on all assessment days. Post-treatment, treatment-related diarrhea was reported in six dogs receiving Credelio Quattro and one dog receiving Credelio on Day 0, with all dogs recovering on Day 1. Conclusions The laboratory studies described confirm the safety and effectiveness of a single dose of Credelio Quattro and Credelio, at the minimum effective dosage of 20 mg/kg lotilaner, 0.02 mg/kg moxidectin, 5 mg/kg praziquantel, 5 mg/kg pyrantel, and 20 mg/kg lotilaner, respectively, for the treatment and control of H. longicornis infestations in dogs for one month. Graphical abstract

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

Borrelia burgdorferi, the causative agent of Lyme disease, is a zoonotic vector-borne pathogen transmitted by various Ixodes tick species. Lyme disease, while commonly asymptomatic, can induce fever and intermittent lameness in dogs. Highly effective acaricidal products with a rapid onset of action along with prompt removal of attached ticks are important aspects of successful Lyme disease prevention strategies. Two studies were conducted with a total of 30 dogs each. Dogs were randomized to receive a control sham dose, Credelio Quattro, or Credelio. Treatment was administered on Day 0 in a fed state. On Day 28, all dogs were experimentally infested with wild-caught adult Ixodes scapularis. Blood samples for B. burgdorferi antibody analysis utilizing the SNAP 4Dx Plus and Lyme Quant C.sub.6 tests were collected on Days 27, 49, 63, 77, 91, and 105. Skin biopsies were collected from four different areas of heavy tick attachment from each dog for polymerase chain reaction (PCR) detection of B. burgdorferi on Day 104 or 105. All control dogs demonstrated adequate I. scapularis infestation rates on Day 33 in both studies. In Study 2, on Day 27, one control dog tested positive for B. burgdorferi on the Lyme Quant C.sub.6 test, prior to experimental tick infestation, and therefore was excluded from analysis. A total of eight out of 10 (Study 1) and nine out of nine (Study 2) control dogs tested positive for B. burgdorferi on at least one test after Day 27. One dog in the Credelio Quattro-treated group tested positive for B. burgdorferi on SNAP 4Dx Plus on Day 105 in Study 1 but was negative on all other tests and study days. None of the dogs treated with Credelio tested positive for B. burgdorferi at any point during either study. The laboratory studies described herein confirm that a single dose of lotilaner, at the minimum effective dosage of 20 mg/kg, administered as Credelio Quattro, in combination with moxidectin, praziquantel, and pyrantel or Credelio, is effective for the prevention of transmission of B. burgdorferi from infected I. scapularis for a full month in dogs.

Haemaphysalis longicornis, the longhorned tick, is an invasive tick species that has been identified in increasing numbers and regions across the USA. This tick species is a competent vector for various pathogens to dogs, humans, and other species, with heavy infestations documented to lead to exsanguination. Therefore, determination of ectoparasiticides providing adequate treatment and control of H. longicornis is imperative to help reduce vector-borne disease transmission and protect against infestation in dogs. Three laboratory studies were conducted to evaluate the efficacy of Credelio Quattro and Credelio for the treatment and control of H. longicornis. A total of 30 dogs per study were randomized to receive either placebo, Credelio Quattro, or Credelio on Day 0 according to a complete block design on the basis of pre-treatment live attached H. longicornis counts, infested on Day -7. To assess efficacy against preexisting infestations, enrolled dogs were infested with 50 unfed adult H. longicornis on Day -2 or -1 prior to treatment. Residual efficacy post-treatment was evaluated with subsequent infestations on Days 5, 12, 19, and 30. All ticks were collected and evaluated as live or dead and free or attached 48 h after treatment or infestation. Adequacy of infestation was achieved in at least two studies for every infestation time point evaluated. Both Credelio Quattro and Credelio provided 100% efficacy against H. longicornis from Day 2 through Day 32, with no live ticks observed on any dogs. A statistically significant number of dead ticks were recovered from both treated groups as compared with control on all assessment days. Post-treatment, treatment-related diarrhea was reported in six dogs receiving Credelio Quattro and one dog receiving Credelio on Day 0, with all dogs recovering on Day 1. The laboratory studies described confirm the safety and effectiveness of a single dose of Credelio Quattro and Credelio, at the minimum effective dosage of 20 mg/kg lotilaner, 0.02 mg/kg moxidectin, 5 mg/kg praziquantel, 5 mg/kg pyrantel, and 20 mg/kg lotilaner, respectively, for the treatment and control of H. longicornis infestations in dogs for one month.