Wild ungulate species differ in their contribution to the transmission of Ixodes ricinus-borne pathogens.

BACKGROUND: Several ungulate species are feeding and propagation hosts for the tick Ixodes ricinus as well as hosts to a wide range of zoonotic pathogens. Here, we focus on Anaplasma phagocytophilum and Borrelia burgdorferi (s.l.), two important pathogens for which ungulates are amplifying and dilution hosts, respectively. Ungulate management is one of the main tools to mitigate human health risks associated with these tick-borne pathogens. Across Europe, different species of ungulates are expanding their ranges and increasing in numbers. It is currently unclear if and how the relative contribution to the life-cycle of I. ricinus and the transmission cycles of tick-borne pathogens differ among these species. In this study, we aimed to identify these relative contributions for five European ungulate species. METHODS: We quantified the tick load and collected ticks and spleen samples from hunted fallow deer (Dama dama, n = 131), moose (Alces alces, n = 15), red deer (Cervus elaphus, n = 61), roe deer (Capreolus capreolus, n = 30) and wild boar (Sus scrofa, n = 87) in south-central Sweden. We investigated the presence of tick-borne pathogens in ticks and spleen samples using real-time PCR. We determined if ungulate species differed in tick load (prevalence and intensity) and in infection prevalence in their tissue as well as in the ticks feeding on them. RESULTS: Wild boar hosted fewer adult female ticks than any of the deer species, indicating that deer are more important as propagation hosts. Among the deer species, moose had the lowest number of female ticks, while there was no difference among the other deer species. Given the low number of infected nymphs, the relative contribution of all ungulate species to the transmission of B. burgdorferi (s.l.) was low. Fallow deer, red deer and roe deer contributed more to the transmission of A. phagocytophilum than wild boar. CONCLUSIONS: The ungulate species clearly differed in their role as a propagation host and in the transmission of B. burgdorferi and A. phagocytophilum. This study provides crucial information for ungulate management as a tool to mitigate zoonotic disease risk and argues for adapting management approaches to the local ungulate species composition and the pathogen(s) of concern.

First report of Theileria annulata in Nigeria: Findings from cattle ticks in Zamfara and Sokoto States.

BACKGROUND: Ticks and tick-borne pathogens (TBPs) represent a significant economic burden to cattle farming in sub-Saharan Africa including Nigeria. However, in the northern part of this country, where the largest livestock population resides, little is known about the contemporary diversity of ticks and TBPs. This area is particularly vulnerable to climate change, undergoing marked transformation of habitat and associated flora and fauna that is also likely to include ticks. This study aimed to document the occurrence of tick species and Apicomplexan TBPs in cattle from north-western Nigeria. METHODS: In 2017, ticks were collected from cattle in Zamfara and Sokoto States and identified morphologically. Additionally, a subset of ticks was screened molecularly for the detection of apicomplexan DNA. RESULTS: A total of 494 adult ticks were collected from 80 cattle in Zamfara and 65 cattle in Sokoto State. Nine tick species were encountered, among which the presence of one, Hyalomma turanicum, had not previously been recorded in Nigeria. Hyalomma rufipes was the most prevalent tick infesting cattle in Zamfara State (76%), while Hyalomma dromedarii was the most prevalent in Sokoto State (44%), confirming the widespread transfer of this species from camels onto livestock and its adaptation to cattle in the region. Of 159 ticks screened, 2 out of 54 (3.7%) from Zamfara State and 29 out of 105 (27.6%) from Sokoto State harboured DNA of Theileria annulata, the agent of tropical theileriosis. CONCLUSIONS: This study confirms the presence of a broad diversity of tick species in cattle from north-western Nigeria, providing the first locality records for Zamfara State. The occurrence of H. turanicum indicates a distribution of this tick beyond northern Africa. This study provides the first report for T. annulata in Nigerian ticks. Given its enormous burden on livestock farming in north Africa and across Asia, further investigations are needed to better understand its epidemiology, vector transmission and potential clinical significance in cattle from northern Nigeria and neighbouring Sahelian countries.

Why Lyme disease is common in the northern US, but rare in the south: The roles of host choice, host-seeking behavior, and tick density.

Lyme disease is common in the northeastern United States, but rare in the southeast, even though the tick vector is found in both regions. Infection prevalence of Lyme spirochetes in host-seeking ticks, an important component to the risk of Lyme disease, is also high in the northeast and northern midwest, but declines sharply in the south. As ticks must acquire Lyme spirochetes from infected vertebrate hosts, the role of wildlife species composition on Lyme disease risk has been a topic of lively academic discussion. We compared tick-vertebrate host interactions using standardized sampling methods among 8 sites scattered throughout the eastern US. Geographical trends in diversity of tick hosts are gradual and do not match the sharp decline in prevalence at southern sites, but tick-host associations show a clear shift from mammals in the north to reptiles in the south. Tick infection prevalence declines north to south largely because of high tick infestation of efficient spirochete reservoir hosts (rodents and shrews) in the north but not in the south. Minimal infestation of small mammals in the south results from strong selective attachment to lizards such as skinks (which are inefficient reservoirs for Lyme spirochetes) in the southern states. Selective host choice, along with latitudinal differences in tick host-seeking behavior and variations in tick densities, explains the geographic pattern of Lyme disease in the eastern US.

Ixodes scapularis (Acari: Ixodidae) Nymphal Survival and Host-Finding Success in the Eastern United States.

The blacklegged tick (Ixodes scapularis Say) is the primary vector of Borrelia burgdorferi sensu stricto (Spirochaetales: Spirochaetaceae), the Lyme disease agent in North America. The basic reproduction number (R0) for B. burgdorferi in I. scapularis in the Northeast is highly sensitive to the probability that engorged larvae survive the winter, molt into nymphs, and find a host. These processes are dependent on local environmental variables, including climate, host population size and movement, and tick behavior. A simple model is presented for estimating host-finding success from the ratio of tick abundance in two subsequent years, accounting for overwinter survival and possible differences in host associations between nymphs and larvae. This model was parameterized using data from two sites in mainland Connecticut and two on Block Island, RI. Host abundance and tick burdens were estimated via mark-recapture trapping of the primary host, Peromyscus leucopus Rafinesque. Overwintering survival was estimated using engorged larvae placed in field enclosures at each site. Only nymphs were recovered alive, and no significant differences in model parameters were observed between Connecticut and Block Island. Host-finding success was predicted to be high across a wide range of host association patterns at three of four sites. Assuming equivalent host association between larvae and nymphs, R0 was also estimated to be greater than one at three of four sites, suggesting these conditions allow for the persistence of B. burgdorferi. The model output was highly sensitive to differences between nymphal and larval host associations.

Benefits and Drawbacks of Citizen Science to Complement Traditional Data Gathering Approaches for Medically Important Hard Ticks (Acari: Ixodidae) in the United States.

Tick-borne diseases are increasing in North America. Knowledge of which tick species and associated human pathogens are present locally can inform the public and medical community about the acarological risk for tick bites and tick-borne infections. Citizen science (also called community-based monitoring, volunteer monitoring, or participatory science) is emerging as a potential approach to complement traditional tick record data gathering where all aspects of the work is done by researchers or public health professionals. One key question is how citizen science can best be used to generate high-quality data to fill knowledge gaps that are difficult to address using traditional data gathering approaches. Citizen science is particularly useful to generate information on human-tick encounters and may also contribute to geographical tick records to help define species distributions across large areas. Previous citizen science projects have utilized three distinct tick record data gathering methods including submission of: 1) physical tick specimens for identification by professional entomologists, 2) digital images of ticks for identification by professional entomologists, and 3) data where the tick species and life stage were identified by the citizen scientist. We explore the benefits and drawbacks of citizen science, relative to the traditional scientific approach, to generate data on tick records, with special emphasis on data quality for species identification and tick encounter locations. We recognize the value of citizen science to tick research but caution that the generated information must be interpreted cautiously with data quality limitations firmly in mind to avoid misleading conclusions.

A combined transcriptomic approach to identify candidates for an anti-tick vaccine blocking B. afzelii transmission.

Ixodes ricinus is the vector for Borrelia afzelii, the predominant cause of Lyme borreliosis in Europe, whereas Ixodes scapularis is the vector for Borrelia burgdorferi in the USA. Transcription of several I. scapularis genes changes in the presence of B. burgdorferi and contributes to successful infection. To what extend B. afzelii influences gene expression in I. ricinus salivary glands is largely unknown. Therefore, we measured expression of uninfected vs. infected tick salivary gland genes during tick feeding using Massive Analysis of cDNA Ends (MACE) and RNAseq, quantifying 26.179 unique transcripts. While tick feeding was the main differentiator, B. afzelii infection significantly affected expression of hundreds of transcripts, including 465 transcripts after 24 h of tick feeding. Validation of the top-20 B. afzelii-upregulated transcripts at 24 h of tick feeding in ten biological genetic distinct replicates showed that expression varied extensively. Three transcripts could be validated, a basic tail protein, a lipocalin and an ixodegrin, and might be involved in B. afzelii transmission. However, vaccination with recombinant forms of these proteins only marginally altered B. afzelii infection in I. ricinus-challenged mice for one of the proteins. Collectively, our data show that identification of tick salivary genes upregulated in the presence of pathogens could serve to identify potential pathogen-blocking vaccine candidates.

An exploratory analysis of demography and movement patterns of dogs: New insights in the ecology of endemic Rocky Mountain-Spotted Fever in Mexicali, Mexico.

An outbreak of Rocky Mountain spotted fever (RMSF) transmitted by the brown dog tick (Rhipicephalus sanguineus sensu lato) has emerged as a major human and animal health concern in Mexicali, Mexico. Due to high rates of brown dog tick infestation, susceptibility, and association with humans, dogs serve as sentinels and have a key role in the ecology of RMSF. A cross-sectional household questionnaire study was conducted in six rural and urban locations to characterize dog ecology and demography in RMSF high-and low-risk areas of Mexicali. In addition, we tracked movement patterns of 16 dogs using a GPS data logger. Of 253 households, 73% owned dogs, and dog ownership tended to be higher in high-risk areas, with a mean dog:human ratio of 0.43, compared with 0.3 in low-risk areas. Dogs in high-risk areas had higher fecundity and roamed more, but the dog density and numbers of free-roaming dogs were comparable. There was a higher proportion of younger dogs and lower proportion of older dogs in high-risk areas. The high proportion of immunologically naïve puppies in high risk areas could result in a lack of herd immunity leading to a more vulnerable dog and human population. The marked increase of space use of free-roaming dogs in high-risk areas suggests that unrestrained dogs could play an important role in spreading ticks and pathogens. As means to limit RMSF risk, practical changes could include increased efforts for spay-neuter and policies encouraging dog restraint to limit canine roaming and spread of ticks across communities; due to dog density is less impactful such policies may be more useful than restrictions on the number of owned dogs.

Experimental infection of Monodelphis domestica with Rickettsia parkeri.

This study aimed to evaluate the infection of the marsupial Monodelphis domestica (the gray short-tailed opossum) by Rickettsia parkeri and its role as an amplifier of the bacterium for Amblyomma ticks. Ten M. domestica males were inoculated with phosphate-buffered saline (PBS) and 106 Vero cells infected with R. parkeri. In seven animals, inoculation was intramuscular, and in three intraperitoneal. One male (control) received 1 ml of the same vehicle used for inoculation intraperitoneally. The three animals inoculated intraperitoneally were infested with uninfected A. sculptum larvae and nymphs between the 2nd and 9th day post-infection (DPI). Parasitemia was monitored from the 3rd to 9th DPI by polymerase chain reaction, using primers for 17 kDa and ompA. The animals were also clinically evaluated. Of the animals infected intramuscularly, only one was blood-positive by the 5th DPI. The three animals infected intraperitoneally were blood-positive on the 2nd, 5th, 7th, and 9th DPI. Of the ten pools of recovered engorged ticks, six had positive bands. The kidney, liver, heart, and spleen of an intramuscularly infected animal were also positive. The rectal temperature of the animals tested increased only in the first three DPI. The animals inoculated intraperitoneally showed prostration, bristled hair, and weight loss. The study found that R. parkeri was capable of infecting M. domestica, which developed rickettsemia and caused infection in xenodiagnostic ticks.

An individual-based model for the dispersal of Ixodes scapularis by ovenbirds and wood thrushes during fall migration.

Ixodes scapularis is responsible for the transmission of a variety of pathogens in North America, including Borrelia burgdorferi sensu stricto, Anaplasma phagocytophilum and Babesia microti. Songbirds have previously been described as agents of tick dispersal, and a combination of empirical data and modeling efforts have implicated songbirds in the range expansion of I. scapularis northward into Canada during spring bird migration. The role of fall bird migration has received comparatively less attention, particularly at a continental scale. The aim of the current research was to use a novel individual-based modeling approach (IBM) to investigate the role of southward migrating songbirds in the dispersal of I. scapularis within the continental United States. The IBM used in this research explicitly models dispersal by two extensively studied migrating songbird species, wood thrush Hylocichla mustelina and ovenbird Seiurus aurocapillus. Our IBM predicts the annual dispersal of more than four million ticks by H. mustelina and S. aurocapillus, notably into areas as far west as the Dakotas, and as far south as Central Alabama. Predicted dispersal locations include areas where the southern phenotype of I. scapularis dominates, suggestive of a possible mechanism for previously described unidirectional gene flow from north to south. In addition, the model demonstrates that three species-specific songbird traits - breeding range, migration timing, and propensity for tick attachment - each play a major role in the relative magnitude of tick dispersal by different songbird species. The pattern of I. scapularis dispersal predicted by this model suggests that migrating songbirds may have contributed to the range expansion of the tick historically, and may continue to do so presently and into the future, particularly as climate changes the geographic areas that are suitable for I. scapularis. Ultimately, widespread tick dispersal by migrating songbirds likely increases the human risk of Lyme disease and other tick-borne diseases in the United States.

Probability of Rhipicephalus microplus introduction into farms by cattle movement using a Bayesian Belief Network.

Attempts to eliminate Rhipicephalus microplus from Uruguay have been unsuccessful, and, currently, the country is divided into two areas: a tick-free area and a tick-infested area. In the tick-infested area, different farms face different situations. Some farms are in regions where, due to environmental conditions or a lack of infrastructure, it is difficult to eliminate R. microplus, and the only option is to control it. In contrast, other farms can attempt complete removal. Before deciding whether a farmer should attempt to eliminate R. microplus, the probability of reintroduction must be evaluated. The objective of this study was to develop a probabilistic model based on a Bayesian Belief Network (BBN) to assess the likelihood of a farm becoming infested with R. microplus via the introduction of tick-infested cattle. Only the tick-infested area was considered in the development of this model. Nine variables related to environmental conditions and biosecurity measures, with a focus on cattle movement, were considered. Three different sources of data were used to populate the BBN model: data from the literature; a representative national survey from 2016; and a survey developed to identify biosecurity practices on farms. Model sensitivity and specificity were assessed, and an overall accuracy of 92% was obtained. The model was applied to 33 farms located in the tick-infested area. For one farm, the probability of introduction of R. microplus was 1%; for three farms, the probability was between 21% and 34%; for seven farms, it was between 66% and 76%; and for 22 farms, the probability was greater than 83%. This model was useful for estimating the probability of the introduction of R. microplus into farms, making it possible to assess the impact that the evaluated biosecurity measures have on the probability of introduction and, thus, guiding more objective decision making about the control or elimination of R. microplus from farms.

A high gene flow in populations of Amblyomma ovale ticks found in distinct fragments of Brazilian Atlantic rainforest.

The genetic structure of populations of the tick Amblyomma ovale from five distinct areas of the Brazilian Atlantic rainforest was evaluated via DNA sequencing and associated with the presence of domestic dogs acting as hosts at the edge of forest fragments. Ticks were collected from domestic dogs and from the environment between 2015 and 2017. Four collection areas were located in the surroundings and within the Serra do Mar State Park, São Paulo State (23°37'21"S, 45°24'43"W), where dogs were bimonthly monitored along 2 years using camera traps and GSM trackers. To determine the spatial limits of genetic structure, ticks collected upon dogs living near the Serra do Baturié, Ceará State (4°15'40"S, 38°55'54"W) were included as well. A total of 39 haplotypes of 16S rRNA and Cox 1 mitochondrial genes sequences were observed, with 27 of them coming from areas within the Serra do Mar State Park. No haplotype was shared between the Serra do Mar and the Serra do Baturié indicating isolation of tick populations at the scale of 2000 km. Although three different haplotype lineages of A. ovale occurred within the Serra do Mar State Park, no genetic structure was found across the study sites within this park, suggesting high tick gene flow across a range of 45 km. Monitoring data from domestic dogs and wild carnivores showed that these species share the same habitats at the forest edge, with dogs playing a likely limited role in tick dispersal. Our findings have important implications for understanding the genetic structure of wide spread A. ovale along Brazilian rainforest remnants, which can further be associated to tick-borne infectious agents, such as Rickettsia parkeri, and used for predicting future patterns of tick diversity in the Brazilian Atlantic rainforest.

Distribution, Host-Seeking Phenology, and Host and Habitat Associations of Haemaphysalis longicornis Ticks, Staten Island, New York, USA.

Haemaphysalis longicornis, an invasive Ixodid tick, was recently reported in the eastern United States. The emergence of these ticks represents a potential threat for livestock, wildlife, and human health. We describe the distribution, host-seeking phenology, and host and habitat associations of these ticks on Staten Island, New York, a borough of New York City.

The importance of wildlife in the ecology and epidemiology of the TBE virus in Sweden: incidence of human TBE correlates with abundance of deer and hares.

BACKGROUND: Tick-borne encephalitis (TBE) is one tick-transmitted disease where the human incidence has increased in some European regions during the last two decades. We aim to find the most important factors causing the increasing incidence of human TBE in Sweden. Based on a review of published data we presume that certain temperature-related variables and the population densities of transmission hosts, i.e. small mammals, and of primary tick maintenance hosts, i.e. cervids and lagomorphs, of the TBE virus vector Ixodes ricinus, are among the potentially most important factors affecting the TBE incidence. Therefore, we compare hunting data of the major tick maintenance hosts and two of their important predators, and four climatic variables with the annual numbers of human cases of neuroinvasive TBE. Data for six Swedish regions where human TBE incidence is high or has recently increased are examined by a time-series analysis. Results from the six regions are combined using a meta-analytical method. RESULTS: With a one-year time lag, the roe deer (Capreolus capreolus), red deer (Cervus elaphus), mountain hare (Lepus timidus) and European hare (Lepus europaeus) showed positive covariance; the Eurasian elk (moose, Alces alces) and fallow deer (Dama dama) negative covariance; whereas the wild boar (Sus scrofa), lynx (Lynx lynx), red fox (Vulpes vulpes) and the four climate parameters showed no significant covariance with TBE incidence. All game species combined showed positive covariance. CONCLUSIONS: The epidemiology of TBE varies with time and geography and depends on numerous factors, i.a. climate, virus genotypes, and densities of vectors, tick maintenance hosts and transmission hosts. This study suggests that the increased availability of deer to I. ricinus over large areas of potential tick habitats in southern Sweden increased the density and range of I. ricinus and created new TBEV foci, which resulted in increased incidence of human TBE. New foci may be established by TBE virus-infected birds, or by birds or migrating mammals infested with TBEV-infected ticks. Generally, persistence of TBE virus foci appears to require presence of transmission-competent small mammals, especially mice (Apodemus spp.) or bank voles (Myodes glareolus).

Frequent Exposure to Many Hunting Dogs Significantly Increases Tick Exposure.

Certain professionals have more exposure to animals and therefore an increased risk of zoonoses. Professional hunting dog caretakers work with upwards of 50 dogs and are exposed to zoonoses through exposure to multiple potentially infectious canine secretions or excretions, as well as to the ticks that dogs carry. Dog caretakers reported having found embedded ticks on their bodies 5.83 times more than environment-only controls. Zoonotic Lyme disease, first in the United States for morbidity due to a vector-borne infection, has dramatically expanded its geographic range over the last two decades. This finding emphasizes the increased risk of tick-borne diseases, including Lyme disease, based on dog exposure and in areas of disease emergence.

Infestation of Rhipicephalus sanguineus sensu lato on cats in Malta.

Studies on cats as hosts of Rhipicephalus sanguineus sensu lato (s.l.) are scarce. Cats are regarded as infrequent hosts of this species complex, and usually only when dogs are also present. In order to compare the occurrence of developmental stages and mitochondrial DNA haplotypes of R. sanguineus s.l. on cats and other domestic or synanthropic mammalian species, 540 ticks were collected from cats, dogs, hedgehogs and one goat. Collections were made from April to September in 2016 and 2017, from 20 locations in Malta in southern Europe. The sampling sites included six cat colonies, where no dogs were present. Compared to adults, significantly more immatures of R. sanguineus s.l. were found on cats (123 larvae and nymphs versus 10 adults) than on dogs (190 larvae and nymphs versus 173 adults). Furthermore, compared to nymphs, significantly more larvae of R. sanguineus s.l. were found on cats (50 larvae versus 73 nymphs) than on dogs (11 larvae versus 179 nymphs). Adult ticks predominated on male dogs (42 adults versus 28 larvae or nymphs), whereas immatures were significantly more abundant compared to adult ticks on female dogs (142 larvae or nymphs versus 80 adults). Similarly, immature as compared with adult ticks were significantly more likely to occur on female cats (72 immature ticks versus 1 adult) in comparison with male cats (46 immature ticks versus 8 adults). Moreover, R. sanguineus s.l. larvae were found significantly more frequently as compared with nymphs on female cats (38 larvae versus 34 nymphs) than on male cats (12 larvae versus 34 nymphs). To confirm morphological identification and to compare mitochondrial markers (cytochrome c oxidase subunit 1 [cox1] and 16S rRNA genes) of ticks across hosts, 57 ticks were analysed. The amplified parts of the cox1 and 16S rRNA genes of R. sanguineus s.l. collected from various hosts showed 100% sequence identity with each other and with those in GenBank from the middle to western Mediterranean Basin. In conclusion, the present study highlights that cats can be important hosts of the immature life stages of R. sanguineus s.l., even in the absence of dogs. This finding has veterinary-medical significance, because stray cats and free-roaming cats may transport immature stages of R. sanguineus s.l. into gardens, i.e. near dogs and humans.

Implication of Nilgai Antelope (Artiodactyla: Bovidae) in Reinfestations of Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) in South Texas: A Review and Update.

The Cattle Fever Tick Eradication Program was the first parasite eradication program of veterinary importance in the United States and is considered to be one of the greatest disease eradication programs of all time. The program's utilization of pasture vacation and dipping of cattle in acaricide has been extremely successful for controlling Rhipicephalus (Boophilus) microplus (Canestrini) and R. (B.) annulatus (Say), collectively known as cattle fever ticks, on cattle along the Texas border with Mexico for decades. However, the increase of white-tailed deer, Odocoileus virginianus (Zimmermann), populations in South Texas over the last 50 yr has compromised the success of the program. R. (B.) microplus and R. (B.) annulatus infestation data have confirmed that O. virginianus can support the maintenance and movement of both species of cattle fever tick within the permanent quarantine or buffer zone in South Texas along the Rio Grande, and also in the cattle fever tick-free area north and east of the buffer zone. Over the last two decades, increasing populations of exotic nilgai antelope, Boselaphus tragocamelus (Pallas), in South Texas have further complicated cattle fever tick eradication efforts. Historical cattle fever tick infestation data, host source data, and geographical data support the continued role of O. virginianus in maintaining reinfestations of R. (B.) microplus and R. (B.) annulatus in South Texas as well as the increasing role of nilgai antelope in cattle fever tick maintenance and dispersal.

Two randomized, controlled studies to assess the efficacy and safety of lotilaner (Credelio™) in preventing Dermacentor reticulatus transmission of Babesia canis to dogs.

BACKGROUND: Dogs worldwide are at risk of Babesia spp. infections. Preventive efficacy of lotilaner tablets (Credelio™, Elanco) against Babesia canis was evaluated in two studies. METHODS: Sixteen dogs in Study 1 and 12 dogs in Study 2, all seronegative and polymerase chain reaction (PCR) negative for B. canis, were randomized to a sham-treated control group or a lotilaner (20-43 mg/kg) treatment group, administered on Day 0 (Study 1: n = 8/group; Study 2: n = 6/group). Dogs were each infested with 50 Dermacentor reticulatus, a percentage of which (Study 1: 8.0-30.0%; Study 2: 12.2%) were infected with B. canis, in Study 1 on Days 2, 7, 14, 21 and 28, and in Study 2 on Day 28. Ticks were removed and counted on Day 30 in Study 1, and Day 34 in Study 2. Blood was collected for Babesia detection via smear, PCR and immunofluorescence assay (IFA) in Study 1 on Day 2, then approximately weekly through Day 56, and in Study 2 at weekly intervals between Days 28 to 49, and on Days 63 and 91. Additional samples were collected from dogs with body temperature > 39.4 °C (measured three times weekly, from Days 7 to 56 in Study 1 and from Days 35 to 56 in Study 2) and positive for B. canis on blood smear. Dogs with confirmed infections were rescue-treated, removed from the study and, in Study 1, replaced. RESULTS: Across both studies B. canis infection of ticks ranged between 8.0-30.0%. In Study 1, all control dogs were positive for B. canis on blood smear and PCR on Day 10 and IFA on Day 21; on Day 21 seven of eight replacement control dogs were B. canis-positive; no replacement dogs were B. canis-positive following tick removal on Day 30. In Study 2, all control dogs were B. canis-positive on Day 56. All lotilaner-treated dogs remained B. canis-negative at all assessments in both studies. CONCLUSION: Lotilaner efficacy was 100% in preventing establishment of B. canis infection, despite post-treatment challenge with infected ticks on Days 2, 7, 14, 21 and 28.

Transstadial Transmission of Hepatozoon canis by Rhipicephalus sanguineus (Acari: Ixodidae) in Field Conditions.

This study investigated possible transovarial and transstadial transmission of Hepatozoon canis by Rhipicephalus sanguineus (Latreille) ticks collected from naturally infected dogs in a municipal dog shelter and the grounds of the shelter. Four hundred sixty-five engorged nymphs were collected from 16 stray dogs that were found to be infected with H. canis by blood smear and PCR analyses and maintained in an incubator at 28 °C for moulting. Four hundred eighteen nymphs moulted to adults 14-16 d post collection. Unfed ticks from the shelter grounds comprised 1,500 larvae, 2,100 nymphs, and 85 adults; were sorted according to origin, developmental stage, and sex into 117 pools; and screened by 18S rRNA PCR for Hepatozoon infection. Of 60 adult tick pools examined, 51 were infected with H. canis. The overall maximum likelihood estimate (MLE) of infection rate was calculated as 21.0% (CI 15.80-28.21). Hepatozoon canis was detected in 31 out of 33 female pools (MLE 26.96%, CI 17.64-44.33) and 20 out of 27 male pools (MLE 14.82%, CI 20.15-46.41). Among 42 unfed nymph pools collected from the shelter, 26 were infected with H. canis, and MLE of infection was calculated as 1.9% (CI 1.25-2.77). No H. canis DNA was detected in any of the gDNA pools consisting of larva specimens. Partial sequences of the 18S rRNA gene shared 99-100% similarity with the corresponding H. canis isolates. Our results revealed the transstadial transmission of H. canis by R. sanguineus, both from larva to nymph and from nymph to adult, in field conditions. However, there were no evidence of transovarial transmission.

Genetic profiling for Anaplasma and Ehrlichia species in ticks collected in the Eastern Cape Province of South Africa.

BACKGROUND: Anaplasma and Ehrlichia are emerging tick-borne pathogens that cause anaplasmosis and ehrlichiosis in humans and other animals worldwide. Infections caused by these pathogens are deadly if left untreated. There has been relatively no systematic survey of these pathogens among ticks in South Africa, thus necessitating this study. The presence of Anaplasma and Ehrlichia species were demonstrated by PCR in ticks collected from domestic ruminants at some selected communities in the Eastern Cape of South Africa. The ticks were identified by morphological characteristics and thereafter processed to extract bacterial DNA, which was analyzed for the presence of genetic materials of Anaplasma and Ehrlichia. RESULTS: Three genera of ticks comprising five species were identified. The screening yielded 16 positive genetic materials that were phylogenetically related to Ehrlichia sequences obtained from GenBank, while no positive result was obtained for Anaplasma. The obtained Ehrlichia sequences were closely related to E. chaffeensis, E. canis, E. muris and the incompletely described Ehrlichia sp. UFMG-EV and Ehrlichia sp. UFMT. CONCLUSION: The findings showed that ticks in the studied areas were infected with Ehrlichia spp. and that the possibility of transmission to humans who might be tick infested is high.