Invasive Tick Research Articles

Records of the Invasive Cattle Tick Rhipicephalus microplus on Wild Hosts and of a Neotropical Tick, Amblyomma dubitatum, on Invasive Alien Mammals, Southern Brazil.

Invasive species threaten wildlife. We detected invasive ticks on wildlife and indigenous ticks on invasive mammals in southern Brazil. The invasive cattle tick Rhipicephalus microplus was found on 12 native mammals and one invasive exotic (Axis axis). The Neotropical tick Amblyomma dubitatum was found on three invasive alien mammal species.

Hyalomma marginatum - A silent stowaway after vacation at the Adriatic Sea

Hyalomma marginatum is an invasive tick species capable of transmitting pathogens that cause severe diseases such as Crimean-Congo hemorrhagic fever. In Austria, H. marginatum occurs sporadically, and migratory birds are believed to bring H. marginatum from distant regions. We report several incidents of H. marginatum imported by travelers in private cars from Croatia to Austria. One tick was positive for Rickettsia aeschlimannii. Tourist traffic may play a significant role in the northward expansion of this tick species aside from introduction by birds.

Ecological Changes Exacerbating the Spread of Invasive Ticks has Driven the Dispersal of Severe Fever with Thrombocytopenia Syndrome Virus Throughout Southeast Asia.

Severe fever with thrombocytopenia syndrome virus (SFTSV) is a tick-borne virus recognized by the World Health Organization as an emerging infectious disease of growing concern. Utilizing phylodynamic and phylogeographic methods, we have reconstructed the origin and transmission patterns of SFTSV lineages and the roles demographic, ecological, and climatic factors have played in shaping its emergence and spread throughout Asia. Environmental changes and fluctuations in tick populations, exacerbated by the widespread use of pesticides, have contributed significantly to its geographic expansion. The increased adaptability of Lineage L2 strains to the Haemaphysalis longicornis vector has facilitated the dispersal of SFTSV through Southeast Asia. Increased surveillance and proactive measures are needed to prevent further spread to Australia, Indonesia, and North America.

Spatial patterns of Hyalomma marginatum-borne pathogens in the Occitanie region (France), a focus on the intriguing dynamics of Rickettsia aeschlimannii.

Hyalomma marginatum is an invasive tick species recently established in mainland southern France. This tick is known to host a diverse range of human and animal pathogens. While information about the dynamics of these pathogens is crucial to assess disease risk and develop effective monitoring strategies, few data on the spatial dynamics of these pathogens are currently available. We collected ticks in 27 sites in the Occitanie region to characterize spatial patterns of H. marginatum-borne pathogens. Several pathogens have been detected: Theileria equi (9.2%), Theileria orientalis (0.2%), Anaplasma phagocytophilum (1.6%), Anaplasma marginale (0.8%), and Rickettsia aeschlimannii (87.3%). Interestingly, we found a spatial clustered distribution for the pathogen R. aeschlimannii between two geographically isolated areas with infection rates and bacterial loads significantly lower in Hérault/Gard departments (infection rate 78.6% in average) compared to Aude/Pyrénées-Orientales departments (infection rate 92.3% in average). At a smaller scale, R. aeschlimannii infection rates varied from one site to another, ranging from 29% to 100%. Overall, such high infection rates (87.3% on average) and the effective maternal transmission of R. aeschlimannii might suggest a role as a tick symbiont in H. marginatum. Further studies are thus needed to understand both the status and the role of R. aeschlimannii in H. marginatum ticks.IMPORTANCETicks are obligatory hematophagous arthropods that transmit pathogens of medical and veterinary importance. Pathogen infections cause serious health issues in humans and considerable economic loss in domestic animals. Information about the presence of pathogens in ticks and their dynamics is crucial to assess disease risk for public and animal health. Analyzing tick-borne pathogens in ticks collected in 27 sites in the Occitanie region, our results highlight clear spatial patterns in the Hyalomma marginatum-borne pathogen distribution and strengthen the postulate that it is essential to develop effective monitoring strategies and consider the spatial scale to better characterize the circulation of tick-borne pathogens.

Attractant and repellent properties of Senna didymobotrya plant extracts to Amblyomma variegatum and Rhipicephalus appendiculatus

The growing challenge of acaricide resistance and geographical range expansion of invasive tick species demands other interventions, like plant-based alternatives, for sustainable tick control. Leaves, flowers, seedpods, and twig branch extracts of Senna didymobotrya were analyzed using coupled gas chromatography mass spectrometry (GC-MS). Response of adult Amblyomma variegatum and Rhipicephalus appendiculatus to extracts was evaluated. The most attractive plant extract was fractionated and ticks’ responses to its fractions assessed. Potential tick attractants in the attractive plant part extract and its fractions were identified by GC-MS analysis. Non- significant qualitative and quantitative differences were observed in the plant parts’ extract composition (R = 0.6178). Flower extracts attracted both species, with a 0.1-fold higher attraction in A. variegatum compared to the standard attraction aggregation attachment pheromone (AAAP). Leaf and seedpod extracts repelled ticks at various concentrations. Bioassays after fractionating flower extracts identified hexane and ethyl acetate fractions as most attractive to A. variegatum (P < 0.001) and R. appendiculatus (P < 0.001), respectively. Chemical analysis of the most attractive extracts and fractions identified compounds, including documented acarine attractants, squalene and linoleic acid. A squalene and linoleic acid blend (1:1) at 1 mg/mL significantly attracted adult A. variegatum (P < 0.01) and R. appendiculatus (P < 0.001). The results of this study broaden comprehension of how ticks respond to plants in nature, and showcase the promising potential for integrating these insights into effective tick management programs.

Spatially limited pathogen pollution in an invasive tick and host system

Expansion of global commerce has facilitated pathogen pollution via the transportation and translocation of invasive species and their associated parasites and pathogens. In Florida, imported cane toads (Rhinella horribilis) were accidentally and intentionally released on multiple occasions. Early populations were found to be infested with the invasive tick, Amblyomma rotundatum, yet it is unknown if these ticks dispersed with their hosts as cane toads spread throughout much of the state. The objectives of our investigation were to (1) determine if there are fewer tick infestations on toads at the periphery than at the core of their distribution as predicted by founder effect events, and (2) identify if ticks were infected with exotic pathogens. We captured toads from 10 populations across Florida. We collected ticks, vent tissue, and tick attachment site tissue from each toad, then tested samples for bacteria in the genus, Rickettsia. We found that 3/10 populations had toads that were infested with A. rotundatum, and infested individuals were in the earliest introduced populations at the core of their distribution. Pathogen testing confirmed Rickettisa bellii in ticks, but not in toad tissues. Haplotype networks could not clearly distinguish if R. bellii in Florida was more closely related to North or South American strains, but host-tick associations suggest that the pathogen was exotic to Florida. Our investigation demonstrated that an invasive species facilitated the introduction of parasites and pathogens into Florida, yet the invasive tick species encountered limitations to dispersal on this host species.

Population genetics of the invasive tick &lt;i&gt;Rhipicephalus microplus&lt;/i&gt; in Benin and Burkina Faso (West Africa)

Rhipicephalus microplus, is an invasive tick species associated with the highest economic losses where it occurs. Invasion of West Africa, where breeding is in 95% of case with low input, started in 2000s and was followed by breeder’s complaints of acaricide resistance. Since understanding of population structure could help tick control, this study aimed to investigate processes which influence R. microplus invasion in Benin and Burkina Faso. Thus, seven microsatellites (SSRs) markers were applied to analyse 436 ticks from Benin and Burkina Faso. Subsequently, determination of population limits, population size, and investigation on isolation by distance pattern were achieved. Analyses revealed that herd is the relevant level of population limit in Benin and with the whole dataset. Significant differentiation was highlighted between herds and between R. microplus population from Benin and Burkina Faso. Migration occurred between herds, as indicated by assignment results and migration rates. Furthermore, any bottleneck was not evidenced within dataset. Results suggested that the origin of R. microplus population of Burkina Faso could be Côte d’Ivoire. The limit of population being the herd has obviously to be considered in tick control strategies. Thus, to be efficient, tick control programs should primarily focus on the cattle coming back north after transhumance.

Molecular identification of cattle ticks in the Greater Accra Region of Ghana: a high occurrence of Rhipicephalus microplus.

Ticks are competent vectors of a wide range of pathogens. They are of veterinary and public health importance as they affect both animal and human health. Transhumance and the transboundary movements of cattle within the West African Sub-region have facilitated the spread of ticks which threatens the introduction of invasive species. Currently, Rhipicephalus microplus have been identified in the Upper East Region of Ghana which could mean a wider distribution of the species across the country due to livestock trade. This study focused on three sites in the Greater Accra Region, which serves as the gateway to receiving most of the cattle transported from the northern regions of Ghana. Ticks were sampled from August 2022 in the wet season to January 2023 in the dry season. Three tick genera were identified: Amblyomma (19.5%), Hyalomma (1.1%), and Rhipicephalus (79.3%) from the 1,489 feeding ticks collected from cattle. Furthermore, Rhipicephalus microplus, Hyalomma rufipes and Amblyomma variegatum were identified molecularly using primers that target the mitochondrial COI gene. There was a significant association between the tick species and seasons (p < 0.001). Finding R. microplus in this study indicates the extent of the spread of this invasive tick species in Ghana and highlights the need for efficient surveillance systems and control measures within the country.

Effect of short photoperiod on the development of Haemaphysalis longicornis (Ixodida: Ixodidae).

The invasive tick, Haemaphysalis longicornis Neumann, is now present across most of the mid-Atlantic States in the eastern United States. This tick ends its seasonal activity in late October to early November, with larvae being the last life-stage observed questing. Previous research has revealed that the activity of H. longicornis is influenced by photoperiod: short daylight lengths trigger diapause in nymphs, marking it as the primary overwintering stage. However, whether engorged larvae can enter diapause or are affected by short daylight is unclear. In this study, we tested in the laboratory whether the photoperiod Affects the development of H. longicornis engorged larvae and engorged nymphs under constant temperature and humidity. The results showed that engorged larvae molted significantly faster (3 days faster) when the photoperiod was 9 h of light as opposed to 14 h. In contrast, changes in the photoperiod did not affect the molting of engorged nymphs. Our results demonstrate that engorged larvae respond to short daylight length, by molting faster. These results suggest that engorged larvae are unlikely to overwinter under field conditions and support the expectation that nymphs are the primary overwintering stage for H. longicornis in the United States.

Eco-climate drivers shape virome diversity in a globally invasive tick species.

Spillovers of viruses into human occur more frequently under warmer conditions, particularly arboviruses. The invasive tick species Haemaphysalis longicornis poses a significant public health threat due to its global expansion and its potential to carry a wide range of pathogens. We analyzed meta-transcriptomic data from 3595 adult H. longicornis ticks collected between 2016 and 2019 in 22 provinces across China, encompassing diverse ecological conditions. Generalized additive modelling revealed that climate factors exerted a stronger influence on the virome of H. longicornis compared to other ecological factors, such as ecotypes, distance to coastline, animal host, tick gender, and anti-viral immunity. We investigated the mechanistic understanding of how climate changes drive the tick virome using causality inference and emphasized its significance for public health. Our findings demonstrated that higher temperatures and lower relative humidity/precipitation contribute to variations in animal host diversity, leading to an increased diversity of tick virome, particularly the evenness of vertebrate associated viruses. This finding may explain the evolution of tick-borne viruses into generalists across multiple hosts, thereby increasing the probability of spillover events involving tick-borne pathogens. Deep learning projections indicate that the diversity of H. longicornis virome is expected to increase in 81.9% of regions under the SSP8.5 scenario from 2019-2030. Extension of surveillance should be implemented to avert the spread of tick-borne diseases.

Morphological characterization of ticks of the genus &lt;em&gt;Rhipicephalus (Boophilus)&lt;/em&gt; based on geometric morphometry: the case of the Savannah District in Côte d’Ivoire

The introduction of the invasive tick Rhipicephalus (Boophilus) microplus in Côte d'Ivoire has led to a change in the population of ticks of the genus Rhipicephalus (Boophilus). Thus, this study aims to characterize morphologically the tick species of the genus Rhipicephalus (B.) by geometric morphometry. It consisted of collecting all ticks from cattle in 74 farms in the Savanes District. The collected ticks were identified in the laboratory and, after identification, only non-gorged adult ticks of the genus Rhipicephalus (B.) were retained for morphometric characterisation. A total of 394 ticks were examined under a digital microscope, images were digitised using the online program XYOM and the data were analysed. This study revealed that males of each species have a smaller average size than females. It also showed a variation in mean size in all species except for the females of Rhipicephalus (B.) decoloratus and Rhipicephalus (B.) geigyi whose mean sizes did not differ. Geometric morphometry allowed the separation of Rhipicephalus (B.) annulatus males from other males. In females, Rhipicephalus (B.) geigyi and Rhipicephalus (B.) microplus were distinguished from each other and from other females.

Establishment and range expansion of Dermacentor variabilis in the northern Maritimes of Canada: Community participatory science documents establishment of an invasive tick species.

Tick populations are dependent on a complex interplay of abiotic and biotic influences, many of which are influenced by anthropic factors including climate change. Dermacentor variabilis, the wood tick or American dog tick, is a hardy tick species that feeds from a wide range of mammals and birds that can transmit pathogens of medical and agricultural importance. Significant range expansion across North America has been occurring over the past decades;this study documents northwards range expansion in the Canadian Maritime provinces. Tick recoveries from passive surveillance between 2012 and 2021 were examined to assess northward population expansion through Atlantic Canada. At the beginning of this period, D. variabilis was abundant in the most southerly province, Nova Scotia, but was not considered established in the province to the north, New Brunswick. During the 10-year span covered by this study, an increasing number of locally acquired ticks were recovered in discrete foci, suggesting small established or establishing populations in southern and coastal New Brunswick. The pattern of population establishment follows the climate-driven establishment pattern of Ixodes scapularis to some extent but there is also evidence of successful seeding of disjunct populations in areas identified as sub-optimal for tick populations. Dogs were the most common host from which these ticks were recovered, which raises the possibility of human activity, via movement of companion animals, having a significant role in establishing new populations of this species. Dermacentor variabilis is a vector of several pathogens of medical and agricultural importance but is not considered to be a competent vector for Borrelia burgdorferi, the etiological agent of Lyme disease; our molecular analysis of a subset of D. variabilis for both B. burgdorferi and B. miyamotoi did not confirm any with Borrelia. This study spans the initial establishment of this tick species and documents the pattern of introduction, providing a relatively unique opportunity to examine the first stages of range expansion of a tick species.

Management of Haemaphysalis longicornis (Acari: Ixodidae) on a cow-calf farm in East Tennessee, USA.

Haemaphysalis longicornis Neumann is an exotic and invasive tick species rapidly expanding across the United States. Large infestations of these ticks on cattle can cause anemia, and in eastern regions of the country, they are transmitting Theileria orientalis Ikeda within herds. Methods for managing H. longicornis in the environment are rarely successful and rely on chemicals. To document population effects from producer management decisions, we investigated the change in H. longicornis populations to different tick management practices chosen by producers at 3 different H. longicornis-infested farms. Farm 1 kept a closed herd, chose to cut brush monthly, used on-animal acaricides, and allowed us to drag weekly. Farm 2 kept an open herd, chose to cut brush yearly, did not use on-animal acaricides, and allowed us to drag for ticks weekly/monthly. Farm 3 kept an open herd, chose to cut brush yearly, used on-animal acaracides, and allowed us to drag for ticks monthly. We assessed H. longicornis populations responding to those methods over time using active surveillance methods at each farm. Management decisions on Farms 1 and 3 significantly reduced H. longicornis on farms. Importantly, Farm 1 management decisions reduced the chance of a tick being present by 90%; the number of questing ticks collected fell from 5,000 to 12 H. longicornis in 2 years. Therefore, combined management strategies such as keeping a closed herd, use of on-animal chemical control, mechanical control, and reducing the amount of vegetation on farms were effective ways for producers to manage Haemaphysalis longicornis Neuman (Ixodidae).

Invasive Haemaphysalis longicornis (Acari: Ixodidae) investigation in South Carolina: new records of establishment, pathogen prevalence, and blood meal analyses.

The first established population of the Asian longhorned tick Haemaphysalis longicornis (Neumann, Acari: Ixodidae) was discovered in a northern South Carolina county in June 2022. A coordinated investigation was launched to investigate the invasive tick's pathogen infection prevalence and blood meal preferences. Almost 2,000 Ha. longicornis ticks were collected from one cattle field. A majority of collected ticks had evidence of cattle and dog blood meals, and multiple samples were tested positive for Borrelia burgdorferi s.l. and Theileria orientalis-first reports for these pathogens in this tick species in South Carolina. This investigation was the direct result of a collaborative education campaign and tick surveillance program launched earlier in the year with multiple state partners.

Ticks (Acari: Ixodida) on synanthropic small and medium-sized mammals in areas of the northeastern United States infested with the Asian longhorned tick, Haemaphysalis longicornis

The northeastern United States (US) is a hotspot for tick-borne diseases. Adding to an already complex vector landscape, in 2017 large populations of the invasive Haemaphysalis longicornis, the Asian longhorned tick, were detected in New Jersey (NJ) and later found to be widespread from Connecticut to Georgia. In its native range in northeastern Asia, H. longicornis is considered an important vector of deadly pathogens to humans, companion animals, and livestock. To identify the primary hosts of H. longicornis, we surveyed synanthropic small and medium-sized mammals in three different sites in suburban New Brunswick, NJ. Specifically, we collected approximately 9,000 tick specimens belonging to nine species from 11 different species of mammals sampled between May and September 2021. We found that H. longicornis feeds more frequently on rodents than previously thought, and that this invasive tick is likely exposed to important enzootic and zoonotic pathogens. Overall, we obtained detailed information about the seasonal dynamics and feeding patterns of six tick species common in the northeastern US, Haemaphysalis longicornis, Amblyomma americanum, Dermacentor variabilis, Ixodes scapularis, Ixodes texanus and Ixodes cookei. We found that unlike I. scapularis that feeds on mammals of all sizes, H. longicornis feeds on hosts following the general pattern of A. americanum, favoring larger species such as skunks, groundhogs, and raccoons. However, our survey revealed that unlike A. americanum, H. longicornis reaches high densities on Virginia opossum. Overall, the newly invasive H. longicornis was the most numerous tick species, both on multiple host species and in the environment, raising significant questions regarding its role in the epidemiology of tick-borne pathogens, especially those affecting livestock, companion animals and wildlife. In conclusion, our findings provide valuable insights into the tick species composition on mammalian hosts in NJ and the ongoing national expansion of H. longicornis.

EVIDENCE OF PROTOZOAN AND BACTERIAL INFECTION AND CO-INFECTION AND PARTIAL BLOOD FEEDING IN THE INVASIVE TICK HAEMAPHYSALIS LONGICORNIS IN PENNSYLVANIA.

The Asian longhorned tick, Haemaphysalis longicornis, an invasive tick species in the United States, has been found actively host-seeking while infected with several human pathogens. Recent work has recovered large numbers of partially engorged, host-seeking H. longicornis, which together with infection findings raises the question of whether such ticks can reattach to a host and transmit pathogens while taking additional bloodmeals. Here we conducted molecular blood meal analysis in tandem with pathogen screening of partially engorged, host-seeking H. longicornis to identify feeding sources and more inclusively characterize acarological risk. Active, statewide surveillance in Pennsylvania from 2020 to 2021 resulted in the recovery of 22/1,425 (1.5%) partially engorged, host-seeking nymphal and 5/163 (3.1%) female H. longicornis. Pathogen testing of engorged nymphs detected 2 specimens positive for Borrelia burgdorferi sensu lato, 2 for Babesia microti, and 1 co-infected with Bo. burgdorferi s.l. and Ba. microti. No female specimens tested positive for pathogens. Conventional PCR blood meal analysis of H. longicornis nymphs detected avian and mammalian hosts in 3 and 18 specimens, respectively. Mammalian blood was detected in all H. longicornis female specimens. Only 2 H. longicornis nymphs produced viable sequencing results and were determined to have fed on black-crowned night heron, Nycticorax nycticorax. These data are the first to molecularly confirm H. longicornis partial blood meals from vertebrate hosts and Ba. microti infection and co-infection with Bo. burgdorferi s.l. in host-seeking specimens in the United States, and the data help characterize important determinants indirectly affecting vectorial capacity. Repeated blood meals within a life stage by pathogen-infected ticks suggest that an understanding of the vector potential of invasive H. longicornis populations may be incomplete without data on their natural host-seeking behaviors and blood-feeding patterns in nature.

Baseline Susceptibility of Haemaphysalis longicornis to Organophosphate, Carbamate, and Pyrethroid Acaricides.

The Asian longhorned tick, Haemaphysalis longicornis, continues to expand its range in North America, and synthetic acaricides are likely to play an increasing role in managing this species. Acaricide resistance is common in some tick species that infest livestock. However, baseline acaricide susceptibility has not previously been examined in this invasive tick. We used a standard larval packet test to evaluate the susceptibility of the Asian longhorned tick to acaricides currently or formerly used in tick control: propoxur, carbaryl, bifenthrin, permethrin, and coumaphos. Discriminating concentrations were estimated at 6.5, 27.9, 988, 2242, and 808 ppm, respectively. The LC50 for propoxur, carbaryl, permethrin and coumaphos were compared with data available for other tick species and showed that H. longicornis was more susceptible to propoxur, carbaryl, and coumaphos and had a similar susceptibility to permethrin. The results indicate that resistance to these acaricides is not presently a concern for H. longicornis in the United States. However, responsible integrated management and early detection of resistance can help ensure the long-term efficacy of products used for controlling this tick species. This article is protected by copyright. All rights reserved.

New York State Tick Blitz: harnessing community-based science to understand range expansion of ticks.

Ticks and tick-borne diseases are an immense public health burden in New York State (NYS), United States. Tick species and associated pathogens are expanding into new areas, shifting threats to human, and animal health across the state. For example, the invasive tick, Haemaphysalis longicornis Neumann (Acari: Ixodidae), was first detected in the United States in 2017 and has since been identified in 17 states, including NYS. In addition, Amblyomma americanum (L.) (Acari: Ixodidae) is a native tick thought to be reestablishing historical populations in NYS. We implemented a community-based science project called the "NYS Tick Blitz" to determine the distribution of A. americanum and H. longicornis in NYS. Community volunteers were recruited, provided with education, training, and materials to conduct active tick sampling during a 2-wk period in June 2021. Fifty-nine volunteers sampled 164 sites across 15 counties, for a total of 179 separate collection events and 3,759 ticks collected. The most frequently collected species was H. longicornis, followed by Dermacentor variabilis Say (Acari: Ixodidae), Ixodes scapularis Say (Acari: Ixodidae), and A. americanum, respectively. Through the NYS Tick Blitz collections, H. longicornis was identified for the first time in Putnam County. We conducted pooled pathogen testing on a subset of specimens, with the highest rates of infection detected for pathogens transmitted by I. scapularis, including Borrelia burgdorferi, Anaplasma phagocytophilum, and Babesia microti. Most participants who completed a follow-up survey (n = 23, 71.9%) were promoters of the NYS Tick Blitz and 50% (n = 15) reported that they enjoyed participating in meaningful science.

Seasonal Dynamics of Tick Species in the Ecotone of Parks and Recreational Areas in Middlesex County (New Jersey, USA).

People often use parks and other forested areas for outdoor activities such as hiking and walking their dogs. Areas of primary use are paths or grassy meadows on the edges of the forests that constitute transitional areas between different plant communities (aka ecotones). In this study, we monitored the seasonal dynamics of questing ticks in forest/meadow and forest/path ecotones in five areas in Middlesex County, New Jersey (NJ). We found anthropophilic species such as Ixodes scapularis, Amblyomma americanum, and Dermacentor variabilis coexisting with Haemaphysalis longicornis, an invasive tick species first detected in NJ in 2017. Surveillance was conducted weekly from March to November 2020, and collected ticks were identified. The most abundant tick species was H. longicornis (83%), followed by A. americanum (9%), I. scapularis (7%), and D. variabilis (<1%). The seasonal dynamics of A. americanum and I. scapularis in the ecotone were similar to previous surveys in forest habitats. The presence of anthropophilic ticks, particularly I. scapularis, suggests the need for specific control approaches to target these habitats. In addition, the extraordinarily high numbers of H. longicornis collected in ecotones (1.70 ticks/m2) and frequent reports of this species on dogs highlight the importance of monitoring its expansion due to its potential as a vector of animal and human diseases.

Efficacy of fluralaner chewable tablets (Bravecto®) against Asian longhorned tick (Haemaphysalislongicornis) infestations of dogs

BackgroundThe 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.MethodsDogs 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).ResultsPre-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.ConclusionAt 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