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Haemaphysalis longicornis is a common Ixodida tick species found in temperate areas of Asian countries. An anti-tick assay was conducted on adult female H. longicornis ticks. Plant extract solutions were prepared at concentrations of 50, 25, and 10 mg/mL. Tick survival and mortality were assessed by counting the number of dead and live ticks at 24 h, 48 h, 72 h, and 96 h posttreatment. Out of 11 plant extracts screened, Artemisia judaica extract exhibited the highest potency with 100% mortality (5/5) at 48 h when applied at high and moderate concentrations (50 and 25 mg/mL). Similar results were observed at 96 h for the 10 mg/mL group compared to the untreated ticks. Cleome droserifolia extract demonstrated partial activity with 60% (3/5) and 20% (1/5) mortality at 96 h posttreatment at concentrations of 50 and 25 mg/mL, respectively. Forsskaolea tenacissima extract showed a weak effect with 100% tick mortality (5/5) only at the highest treatment concentration after 96 h. To confirm the activity of A. judaica, trial 2 was conducted. A. judaica demonstrated potency within 48 h in high dose and 72 h in moderate dose, with 100% mortality (15/15) at 96 h posttreatment compared to untreated ticks. The median lethal time 50 (LT50) values were 30.37 h for the high and 55.08 h for the moderate doses. Liquid chromatography‒mass spectrometry was performed on the most potent candidate (A. judaica) to identify its phytochemical components. The results revealed the presence of 9 compounds identified through manual annotation and 74 compounds from the Global Natural Products Social library. These compounds included terpenoids, steroids, phenylpropanoids, flavonoid glycosides, flavonoids, and benzenoids. Camphor was identified as the major component via both approaches. These findings suggest the potential use of A. judaica extract in the future development of acaricidal therapeutics. [ABSTRACT FROM AUTHOR]

Amblyomma americanum, a known vector of multiple tick-borne pathogens, has expanded its geographic distribution across the United States in the past decades. Tick microbiomes may play a role shaping their host's life history and vectorial capacity. Bacterial communities associated with A. americanum may reflect, or enable, geographic expansion and studying the microbiota will improve understanding of tick-borne disease ecology. We examined the microbiota structure of 189 adult ticks collected in four regions encompassing their historical and current geographic distribution. Both geographic region of origin and sex were significant predictors of alpha diversity. As in other tick models, within-sample diversity was low and uneven given the presence of dominant endosymbionts. Beta diversity analyses revealed that bacterial profiles of ticks of both sexes collected in the West were significantly different from those of the Historic range. Biomarkers were identified for all regions except the historical range. In addition, Bray-Curtis dissimilarities overall increased with distance between sites. Relative quantification of ecological processes showed that, for females and males, respectively, drift and dispersal limitation were the primary drivers of community assembly. Collectively, our findings highlight how microbiota structural variance discriminates the western-expanded populations of A. americanum ticks from the Historical range. Spatial autocorrelation, and particularly the detection of non-selective ecological processes, are indicative of geographic isolation. We also found that prevalence of Ehrlichia chaffeensis, E. ewingii, and Anaplasma phagocytophilum ranged from 3.40–5.11% and did not significantly differ by region. Rickettsia rickettsii was absent from our samples. Our conclusions demonstrate the value of synergistic analysis of biogeographic and microbial ecology data in investigating range expansion in A. americanum and potentially other tick vectors as well. [ABSTRACT FROM AUTHOR]

Amblyomma ticks are vectors of both Rickettsia rickettsii and R. parkeri in the Americas, where capybaras (Hydrochoerus hydrochaeris) are the main hosts in urban areas, thus contributing to the transmission of spotted fever. Herein, we studied: (i) the seasonal dynamics and abundance of ticks in areas where capybaras live, (ii) the effect of environmental variables on tick abundance, and (iii) the presence of Rickettsia-infected ticks. Between September 2021 and September 2022, we sampled ticks using cloth-dragging at 194 sites on the shore of Lake Paranoá in Brasília, Brazil. We measured environmental data (season, vegetation type, canopy density, temperature, humidity, and presence or vestige of capybara) at each site. Nymphs and adults were morphologically identified to the species level, and a selected tick sample including larvae was subjected to genotypic identification. We investigated Rickettsia-infected ticks by PCR (gltA, htrA, ompB, and ompA genes) and associations between tick abundance and environmental variables using Generalized Linear Models. A total of 30,334 ticks (96% larvae) were captured. Ticks were identified as Amblyomma, with A. sculptum comprising 97% of the adult/nymphs. Genotype identification of a larval sample confirmed that 95% belonged to A. dubitatum. Seasonal variables showed significant effects on tick abundance. Most larvae and nymphs were captured during the early dry season, while the adults were more abundant during the wet season. Vegetation variables and the presence of capybaras showed no association with tick abundance. Rickettsia parkeri group and R. bellii were identified in A. dubitatum, while A. sculptum presented R. bellii. We conclude that: (i) Amblyomma ticks are widely distributed in Lake Paranoá throughout the year, especially larvae at the dry season, (ii) the abundance of Amblyomma ticks is explained more by climatic factors than by vegetation or presence of capybaras, and (iii) A. dubitatum ticks are potential vectors of R. parkeri in Brasília. [ABSTRACT FROM AUTHOR]

Lyme disease is the most common wildlife-to-human transmitted disease reported in North America. The study of this disease requires an understanding of the ecology of the complex communities of ticks and host species involved in harboring and transmitting this disease. Much of the ecology of this system is well understood, such as the life cycle of ticks, and how hosts are able to support tick populations and serve as disease reservoirs, but there is much to be explored about how the population dynamics of different host species and communities impact disease risk to humans. In this study, we construct a stage-structured, empirically-informed model with host dynamics to investigate how host population dynamics can affect disease risk to humans. The model describes a tick population and a simplified community of three host species, where primary nymph host populations are made to fluctuate on an annual basis, as commonly observed in host populations. We tested the model under different environmental conditions to examine the effect of environment on the interactions of host dynamics and disease risk. Results show that allowing for host dynamics in the model reduces mean nymphal infection prevalence and increases the maximum annual prevalence of nymphal infection and the density of infected nymphs. Effects of host dynamics on disease measures of nymphal infection prevalence were nonlinear and patterns in the effect of dynamics on amplitude in nymphal infection prevalence varied across environmental conditions. These results highlight the importance of further study of the effect of community dynamics on disease risk. This will involve the construction of further theoretical models and collection of robust field data to inform these models. With a more complete understanding of disease dynamics we can begin to better determine how to predict and manage disease risk using these models. [ABSTRACT FROM AUTHOR]

The states of Kansas and Oklahoma, in the central Great Plains, lie at the western periphery of the geographic distributions of several tick species. As the focus of most research on ticks and tick-borne diseases has been on Lyme disease which commonly occurs in areas to the north and east, the ticks of this region have seen little research attention. Here, we report on the phenology and activity patterns shown by tick species observed at 10 sites across the two states and explore factors associated with abundance of all and life specific individuals of the dominant species. Ticks were collected in 2020–2022 using dragging, flagging and carbon-dioxide trapping techniques, designed to detect questing ticks. The dominant species was A. americanum (24098, 97%) followed by Dermacentor variabilis (370, 2%), D. albipictus (271, 1%), Ixodes scapularis (91, <1%) and A. maculatum (38, <1%). Amblyomma americanum, A. maculatum and D. variabilis were active in Spring and Summer, while D. albipictus and I. scapularis were active in Fall and Winter. Factors associated with numbers of individuals of A. americanum included day of year, habitat, and latitude. Similar associations were observed when abundance was examined by life-stage. Overall, the picture is one of broadly distributed tick species that shows seasonal limitations in the timing of their questing activity. [ABSTRACT FROM AUTHOR]

Ticks are arachnid ectoparasites that rank second only to mosquitoes in the transmission of human diseases including bacteria responsible for anaplasmosis, ehrlichiosis, spotted fevers, and Lyme disease among other febrile illnesses. Due to the paucity of data on bacteria transmitted by ticks in Kenya, this study undertook a bacterial metagenomic-based characterization of ticks collected from Isiolo, a semi-arid pastoralist County in Eastern Kenya, and Kwale, a coastal County with a monsoon climate in the southern Kenyan border with Tanzania. A total of 2,918 ticks belonging to 3 genera and 10 species were pooled and screened in this study. Tick identification was confirmed through the sequencing of the Cytochrome C Oxidase Subunit 1 (COI) gene. Bacterial 16S rRNA gene PCR amplicons obtained from the above samples were sequenced using the MinION (Oxford Nanopore Technologies) platform. The resulting reads were demultiplexed in Porechop, followed by trimming and filtering in Trimmomatic before clustering using Qiime2-VSearch. A SILVA database pretrained naïve Bayes classifier was used to classify the Operational Taxonomic Units (OTUs) taxonomically. The bacteria of clinical interest detected in pooled tick assays were as follows: Rickettsia spp. 59.43% of pools, Coxiella burnetii 37.88%, Proteus mirabilis 5.08%, Cutibacterium acnes 6.08%, and Corynebacterium ulcerans 2.43%. These bacteria are responsible for spotted fevers, query fever (Q-fever), urinary tract infections, skin and soft tissue infections, eye infections, and diphtheria-like infections in humans, respectively. P. mirabilis, C. acnes, and C. ulcerans were detected only in Isiolo. Additionally, COI sequences allowed for the identification of Rickettsia and Coxiella species to strain levels in some of the pools. Diversity analysis revealed that the tick genera had high levels of Alpha diversity but the differences between the microbiomes of the three tick genera studied were not significant. The detection of C. acnes, commonly associated with human skin flora suggests that the ticks may have contact with humans potentially exposing them to bacterial infections. The findings in this study highlight the need for further investigation into the viability of these bacteria and the competency of ticks to transmit them. Clinicians in these high-risk areas also need to be appraised for them to include Rickettsial diseases and Q-fever as part of their differential diagnosis. [ABSTRACT FROM AUTHOR]

Crimean-Congo haemorrhagic fever orthonairovirus (CCHFV) is a negative-sense, single-stranded RNA virus with a segmented genome and the causative agent of a severe Crimean-Congo haemorrhagic fever (CCHF) disease. The virus is transmitted mainly by tick species in Hyalomma genus but other ticks such as representatives of genera Dermacentor and Rhipicephalus may also be involved in virus life cycle. To improve our understanding of CCHFV adaptation to its tick species, we compared nucleotide composition and codon usage patterns among the all CCHFV strains i) which sequences and other metadata as locality of collection and date of isolation are available in GenBank and ii) which were isolated from in-field collected tick species. These criteria fulfilled 70 sequences (24 coding for S, 23 for M, and 23 for L segment) of virus isolates originating from different representatives of Hyalomma and Rhipicephalus genera. Phylogenetic analyses confirmed that Hyalomma- and Rhipicephalus-originating CCHFV isolates belong to phylogenetically distinct CCHFV clades. Analyses of nucleotide composition among the Hyalomma- and Rhipicephalus-originating CCHFV isolates also showed significant differences, mainly in nucleotides located at the 3rd codon positions indicating changes in codon usage among these lineages. Analyses of codon adaptation index (CAI), effective number of codons (ENC), and other codon usage statistics revealed significant differences between Hyalomma- and Rhipicephalus-isolated CCHFV strains. Despite both sets of strains displayed a higher adaptation to use codons that are preferred by Hyalomma ticks than Rhipicephalus ticks, there were distinct codon usage preferences observed between the two tick species. These findings suggest that over the course of its long co-evolution with tick vectors, CCHFV has optimized its codon usage to efficiently utilize translational resources of Hyalomma species. [ABSTRACT FROM AUTHOR]

A cross-sectional study was conducted from May 2017 to March 2019. A participatory epidemiological appraisal was used to gain a rapid overview of the range of camel calf health problems and traditional management, a measure of the importance that people place on each of them, and to identify and prioritize economically important diseases in target zones. The most important constraints to camel production were identified to be the widespread prevalence of diseases such as camelpox, contagious ecthyma, calf scour, ticks, and nonspecific pneumonia; poor management and husbandry practices such as restrictive colostrum feeding, lack of concentrate and salt supplementation and inappropriate housing; shortage of feed; and scarce seasonal variation in water. Additionally, the livestock herders not only showed their knowledge of common camel calf diseases for affected organs and symptoms but also indicated the seasonality of disease occurrences with strong agreement (W = 0.899, P< 0.003) among the informants of all focus group discussions. The overall prevalence of mange, tick infestation, and bacteria-induced diarrhea in the study area was found to be 36.3%, 36%, and 74%, respectively. Sarcoptes scabie var. cameli was the only identified mite species from mange-infested calves, while Hyalomma, Rhipicephalus, and Amblyoma were the most commonly identified tick species. Similarly, the overall prevalence of diarrhea was 74% among this about 34.6% was caused by E. coli while 38.9% was affected by Salmonella and E. coli. Therefore, based on these findings, five diseases have been prioritized as the most significant calf diseases in the area (Camelpox, contagious ecthyma, and causes of pneumonia among camel calves). Improving veterinary health infrastructure and capacity, and increasing community awareness on camel health constraints are also recommended to enhance optimal camel calf rearing. [ABSTRACT FROM AUTHOR]

Ixodid ticks, such as Ixodes ovatus and Haemaphysalis flava, are important vectors of tick-borne diseases in Japan, such as Japanese spotted fever caused by Rickettsia japonica. This study describes the Rickettsia infection rates influenced by the population genetic structure of I.ovatus and H. flava along an altitudinal gradient. A total of 346 adult I. ovatus and 243 H. flava were analyzed for the presence of Rickettsia by nested PCR targeting the 17kDA, gltA, rOmpA, and rOmpB genes. The population genetic structure was analyzed utilizing the mitochondrial cytochrome oxidase 1 (cox1) marker. The Rickettsia infection rates were 13.26% in I. ovatus and 6.17% in H. flava. For I. ovatus, the global FST value revealed significant genetic differentiation among the different populations, whereas H. flava showed non-significant genetic differentiation. The cox1 I. ovatus cluster dendrogram showed two cluster groups, while the haplotype network and phylogenetic tree showed three genetic groups. A significant difference was observed in Rickettsia infection rates and mean altitude per group between the two cluster groups and the three genetic groups identified within I. ovatus. No significant differences were found in the mean altitude or Rickettsia infection rates of H. flava. Our results suggest a potential correlation between the low gene flow in I. ovatus populations and the spatially heterogeneous Rickettsia infection rates observed along the altitudinal gradient. This information can be used in understanding the relationship between the tick vector, its pathogen, and environmental factors, such as altitude, and for the control of tick-borne diseases in Japan. [ABSTRACT FROM AUTHOR]

Ticks are important vectors of zoonotic diseases and play a major role in the circulation and transmission of many rickettsial species. The aim of this study was to investigate the carriage of Candidatus Rickettsia tarasevichiae (CRT) in a total of 1168 ticks collected in Inner Mongolia to elucidate the potential public health risk of this pathogen, provide a basis for infectious disease prevention, control and prediction and contribute diagnostic ideas for clinical diseases that present with fever in populations exposed to ticks. A total of four tick species, Haemaphysalis concinna (n = 21), Dermacentor nuttalli (n = 122), Hyalomma marginatum (n = 148), and Ixodes persulcatus (n = 877), were collected at nine sampling sites in Inner Mongolia, China, and identified by morphological and molecular biological methods. Reverse transcription PCR targeting the 16S ribosomal RNA (rrs), gltA, groEL, ompB and Sca4 genes was used to detect CRT DNA. Sequencing was used for pathogen species confirmation. The molecular epidemiological analysis showed that three species of ticks were infected with CRT, and the overall positive rate was as high as 42%. The positive rate of I. persulcatus collected in Hinggan League city was up to 96%, and that of I. persulcatus collected in Hulun Buir city was 50%. The pool positive rates of D. nuttalli and H. marginatum collected in Bayan Nur city and H. concinna collected in Hulun Buir city were 0%, 28% and 40%, respectively. This study revealed the high prevalence of CRT infection in ticks from Inner Mongolia and the first confirmation of CRT detected in H. marginatum in China. The wide host range and high infection rate in Inner Mongolia may dramatically increase the exposure of CRT to humans and other vertebrates. The role of H. marginatum in the transmission of rickettsiosis and its potential risk to public health should be further considered. [ABSTRACT FROM AUTHOR]

Robust tick surveillance enhances diagnosis and prevention of tick-borne pathogens, yet surveillance efforts in the United States are highly uneven, resulting in large surveillance vacuums, one of which spans the state of New Mexico. As part of a larger effort to fill this vacuum, we conducted both active and passive tick sampling in New Mexico, focusing on the southern portion of the state. We conducted active tick sampling using dragging and CO₂ trapping at 45 sites across Hidalgo, Doña Ana, Otero, and Eddy counties between June 2021 to May 2022. Sampling occurred intermittently, with at least one sampling event each month from June to October 2021, pausing in winter and resuming in March through May 2022. We also conducted opportunistic, passive tick sampling in 2021 and 2022 from animals harvested by hunters or captured or collected by researchers and animals housed in animal hospitals, shelters, and farms. All pools of ticks were screened for Rickettsia rickettsii, Rickettsia parkeri, Rickettsia amblyommatis, Ehrlichia ewingii, and Ehrlichia chaffeensis. Active sampling yielded no ticks. Passive sampling yielded 497 ticks comprising Carios kelleyi from pallid bats, Rhipicephalus sanguineus from dogs, mule deer, and Rocky Mountain elk, Otobius megnini from dogs, cats, horses, and Coues deer, Dermacentor parumapertus from dogs and black-tailed jackrabbits, Dermacentor albipictus from domesticated cats, mule deer and Rocky Mountain elk, and Dermacentor spp. from American black bear, Rocky Mountain elk, and mule deer. One pool of D. parumapterus from a black-tailed jackrabbit in Luna County tested positive for R. parkeri, an agent of spotted fever rickettsiosis. Additionally, a spotted fever group Rickettsia was detected in 6 of 7 C. kelleyi pools. Two ticks showed morphological abnormalities; however, these samples did not test positive for any of the target pathogens, and the cause of the abnormalities is unknown. Passive surveillance yielded five identified species of ticks from three domestic and six wild mammal species. Our findings update tick distributions and inform the public, medical, and veterinary communities of the potential tick-borne pathogens present in southern New Mexico. [ABSTRACT FROM AUTHOR]

Dabieshan tick virus (DBTV) is a newly identified arbovirus, first detected in Haemaphysalis longicornis collected from Hubei Province in 2015. It has been confirmed that DBTV is widely distributed in Shandong Province, China. However, its entomological and epidemiological features remain to be further explored, particularly the feasibility of transovarial transmission. Our research tries to explain the possibility of transovarial transmission of DBTV from engorged female ticks to their offspring. All engorged female adult ticks were sampled from domestic sheep and allowed to lay eggs and hatch in appropriate laboratory conditions. All engorged ticks, larvae and unhatched eggs were classified into pools for nucleic acid extraction and DBTV RNA detection. According to the results of qRT-PCR, the positive rate of DBTV was 6.25% (8/128) in engorged female ticks, 3.57% (1/28) in eggs and 5% (3/60) in larvae pools, respectively. Phylogenetic analysis indicated that DBTV isolates from larvae were similar to those from maternal ticks with more than 99.5% homology, and DBTV was relatively conservative in evolution. Our findings are the first to provide molecular evidence of potential transovarial transmission of DBTV among H. longicornis. Nonetheless, the transovarial transmission of DBTV in frequency and proportion occurring in nature deserves further investigation. [ABSTRACT FROM AUTHOR]

Ticks play an important role in transmitting many different emerging zoonotic pathogens that pose a significant threat to human and animal health. In Switzerland and abroad, the number of tick-borne diseases, in particular tick-borne encephalitis (TBE), has been increasing over the last few years. Thus, it remains essential to investigate the pathogen spectrum of ticks to rapidly detect emerging pathogens and initiate the necessary measures. To assess the risk of tick-borne diseases in different regions of Switzerland, we collected a total of 10'286 ticks from rural and urban areas in ten cantons in 2021 and 2022. Ticks were pooled according to species, developmental stage, gender, and collection site, and analyzed using next generation sequencing (NGS) and quantitative polymerase chain reaction (qPCR). The metagenomic analysis revealed for the first time the presence of Alongshan virus (ALSV) in Swiss ticks. Interestingly, the pool-prevalence of ALSV was higher than that of tick-borne encephalitis virus (TBEV). Furthermore, several TBEV foci have been identified and pool prevalence of selected non-viral pathogens determined. [ABSTRACT FROM AUTHOR]

Since the 2017 discovery of established populations of the Asian longhorned tick, (Haemaphysalis longicornis Neumann) in the United States, populations continue to be detected in new areas. For this exotic and invasive species, capable of transmitting a diverse repertoire of pathogens and blood feeding on a variety of host species, there remains a lack of targeted information on how to best prepare for this tick and understand when and where it occurs. To fill this gap, we conducted two years of weekly tick surveillance at four farms in Tennessee (three H. longicornis-infested and one without) to identify environmental factors associated with each questing life stage, to investigate predictors of abundance, and to determine the likelihood of not collecting ticks at different life stages. A total of 46,770 ticks were collected, of which 12,607 H. longicornis and five other tick species were identified. Overall, abundance of H. longicornis were associated with spring and summer seasons, forested environments, relative humidity and barometric pressure, sunny conditions, and in relation with other tick species. The likelihood of not collecting H. longicornis was associated with day length and barometric pressure. Additional associations for different life stages were also identified and included other tick species, climatic variables, and environmental conditions. Here, we demonstrated that environmental variables can be useful to predict the presence of questing H. longicornis and provide ideas on how to use this information to develop a surveillance plan for different southeastern areas with and without infestations. [ABSTRACT FROM AUTHOR]

Background: The study aimed to estimate the burden of ticks and tick-borne diseases (TBDs) among rural cattle-keeping households of the Eastern Cape province of South Africa using Productivity Adjusted Life Years (PALYs). We modified Disability Adjusted Life Year (DALY) equations for humans to PALYs to estimate the societal burden of tick-borne animal diseases. Whilst the World Health Organization has indicated the adaptability of DALYs to assess burden of animal diseases, nothing has been done in this regard. This could be due to several reasons including that the assessment of animal disease burden is often less of a priority compared to human diseases, particularly in low- and middle-income countries where resources may be limited. As a result, less funding and attention may be given to developing and implementing PALYs for animal diseases. Furthermore, technical and conceptual challenges may be associated with applying DALYs equations to animal diseases, such as determining appropriate measures of productivity loss for different types and categories of animals and diseases. This motivated our study, which is focused on modelling the burden of ticks and TBDs in cattle (cows, oxen and bulls) reared in resource-poor settings. Method: We formulated a PALYs approach for cattle populations by adapting the DALYs approach to assess the burden of ticks and TBDs for cattle populations in 20 villages in the Eastern Cape province of South Africa. PALYs is a measurement used to assess the burden of disease in cattle populations, quantifying the years of life lost due to premature mortality and disability. It encompasses years of life lost due to premature mortality (YLL) and years lost due to disability (YLD) caused by health conditions. PALYs provide a comprehensive perspective on the effective number of years lost due to disability and premature death in cattle populations. The PALYs model involves several parameters that are examined to understand their impact on the model's behavior. To illustrate this, we used a structured questionnaire to collect data on parameters that feed into PALY equations. We coded and entered data from questionnaires directly into Statistical Package of Social Sciences (IBM SPSS Version 20) and entered the estimated values of PALY parameters to calculate PALYs equations, which were to estimate the societal disease burden of ticks and TBDs in cattle. PALYs calculations were done in three categories; PALYs without discounting and age weighting, PALYs with only discounting, and PALYs with discounting and age weighting in a practical example to study how these parameters influence the outcomes of the PALYs model. Results: Our results revealed that the years of productivity lost by a cow, bull, and ox that suffered from ticks and TBDs could be estimated at various disability weights. Approximately 26%, 23% and 35% of the productivity years of a cow, ox and bull, respectively, reared by resource-poor livestock owners are lost due to the burden of ticks and TBDs in the Eastern Cape province of South Africa. However, introducing tick control measures reduces the loss to approximately 3%, 2% and 3% of their lifespan productivity, an indication that tick control will save approximately 23%, 21% and 32% of years of the productive life of cows, oxen and bulls, respectively. Therefore, it is evident that using ticks and TBD prevention measures at an early age of cattle will improve cattle productivity and hence the socioeconomic welfare of resource-poor rural farming communities in the Eastern Cape province of South Africa. Conclusion: The findings generated from the PALYs approach are helpful in projections for the future burden of any livestock disease. They may be used as a basis in policy formulation and decision-making by various stakeholders, and hence a priority in animal health economics. We recommend that a classification of livestock diseases of national economic importance should consider both the societal burden (non-monetary) and economic impact instead of the common practice of only considering the economic (monetary) impact. Adding a societal burden measure to existing economic measures provides a holistic understanding of the impact of a disease on society especially in resource-limited settings where the livestock value goes beyond monetary value. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Tularemia and Q fever are endemic diseases in Iran; however, little information is available on the prevalence of the causative agents, Coxiella burnetii and Francisella tularensis, in Iranian ticks. This study investigated C. burnetii and F. tularensis among hard ticks in this country. We collected ticks from livestock and other mammals in Guilan, Mazandaran, Golestan (northern Iran), Kurdistan (western Iran), and West Azerbaijan (northwestern Iran) provinces. Genomic DNA from collected ticks was extracted and screened for C. burnetii and F. tularensis using Real-time PCR. A total of 4,197 ticks (belonging to 12 different species) were collected, and Ixodes ricinus (46.4%), Rhipicephalus turanicus (25%), and Rhipicephalus sanguineus sensu lato (19.1%) were the most collected species. Of 708 pooled tick samples, 11.3% and 7.20% were positive for C. burnetii and F. tularensis, respectively. The genus of Rhipicephalus had the highest (18.3%) C. burnetii infection among the collected tick pools (P<0.001). Furthermore, the most positive pools for F. tularensis belonged to Haemaphysalis spp. (44.4%). Kurdistan had the most significant percentage of C. burnetii-infected ticks (92.5%), and there was a meaningful relationship between the provinces and the infection (P< 0.001). The ticks from Golestan exhibited the highest F. tularensis infection rate (10. 9%), and the infection showed no significant relationship with the provinces (P = 0.19). Ticks collected from grasslands had a higher Coxiella burnetii infection rate than those collected from animals (39.4% vs. 7.9%; p<0.01). However, ticks collected from animal surfaces had a slightly higher rate of Francisella tularensis infection than those collected from grasslands (7.6% vs. 3.9%; p = 0.24). Here, we demonstrated the presence of both pathogens in the north (Guilan, Mazandaran, and Golestan provinces), the west (Kurdistan province), and the northwest (West Azerbaijan province) of Iran. The public health system should pay particular attention to tick bites in veterinary medicine and humans. [ABSTRACT FROM AUTHOR]

The Vietnamese government introduced a change in the minimum tick size for stock trading on 12 September 2016 to improve market quality and reduce trade execution costs. The intended effects of this policy have not been widely investigated in an emerging market such as Vietnam. We use data on trade and quote intraday of all stocks listed on the Ho Chi Minh Stock Exchange for the periods before and after the event, with a one-week break from 12/9/2016 to 18/9/2016, for the market to adapt to the new tick size policy. Findings from this paper confirm that the trading cost is reduced following the change to the smallest tick size. However, this is different for large trades executed at the stock price associated with a larger tick size. Furthermore, the findings are robust with a different sample period. These findings imply that introducing a change in tick size in Vietnam in 2016 is desirable for improving market quality. However, the differentiation of these changes in different ranges of stock prices is not necessarily effective for improving market quality and reducing trade execution costs. [ABSTRACT FROM AUTHOR]

Kyasanur Forest Disease is a tick-borne flavivirus is endemic in the Southern India. The recent expansion and resurgence of sporadic outbreaks in southern parts of country is the most important concern. Although only formalin inactivated vaccine is available for treatment with limited efficacy the early detection and timely identification is a only way to prevent spread of cases. If the disease can be identified prior to infection in humans like in forest areas from ticks and vectors the disease spread supposed to be managed quickly. Here we have standardized a single tube ready to use dry-down probe free real time RT-PCR targeted against virus envelope gene for detection of KFDV infection. The assay was standardized in liquid format first, later it was converted into dry-down format with addition of stabilizers with a similar sensitivity and specificity (10RNA Copies/rxn). The sensitivity was comparable to the most widely used and accepted diagnostic platform i.e. TaqMan qRT-PCR. However as the reported assay here omit the need of probes makes it cost effective and dry-down reagents makes more stability to the developed assay in this study if compare to TaqMan qPCR. The assay was evaluated with KFD positive samples and healthy sample panel which revealed high concordance with TaqMan qRT-PCR. Stability was unaffected by temperature fluctuations during transportation even in cold chain free conditions, thus reduce the maintenance of strict cold storage. These findings demonstrated that the reported assay is convenient with 100% sensitivity and specificity to TaqMan qPCR. Thus this assay has the potential usefulness for diagnosis KFDV for routine surveillance in resource limited laboratory settings omitting the use costly and heat sensitive TaqMan qRT-PCR reagents without compromising the sensitivity and specificity of the diagnosis assay. [ABSTRACT FROM AUTHOR]

Due to recent climatic and land use changes, Canada has experienced changes in tick populations, leading to an increased risk of tick bites and tick-borne pathogen exposure, especially in eastern Canada. Preventative recommendations for companion animals from veterinary professionals include regular use of tick prevention products and tick checks. Tick checks, specifically, should target regions of an animal's body which are deemed to be high risk for tick attachment. However, tick species-specific infestation patterns on dogs and cats are largely understudied, and additional research is needed to help guide targeted tick checks. The objective of this study was to identify tick species-specific infestation patterns on dogs and cats. Ticks were collected for one year (April 2019 –March 2020) from 94 veterinary clinics across Canada as part of the Canadian Pet Tick Survey. All ticks were identified to species, and data on the location of tick attachment were ascertained with each submission. To examine the association between location of attachment (outcome) and tick species (explanatory variable), specifically Ixodes scapularis and Dermacentor variabilis, mixed effects univariable models were built. Two thousand three hundred and six submissions were received from 1925 dogs and 381 cats across Canada. Of these submissions, 1377 comprised Ixodes scapularis, and 620 comprised Dermacentor variabilis. Clear tick species-specific infestation patterns for dogs were present, with I. scapularis being significantly more likely to be found on the shoulders, and D. variabilis more likely to be found on the ears and neck. Dermacentor variabilis was more likely to be found on the cranial aspect of cats' limbs, compared to I. scapularis. Up-to-date information on infestation patterns can be used to inform veterinary professionals and pet owners of common attachment sites based on established ticks in their region and thus conduct targeted tick checks. [ABSTRACT FROM AUTHOR]

Tick-borne zoonotic diseases pose a threat to public health; hence, identifying the pathogenic agents associated with them is critical. The prevalence of Bartonella and Rickettsia in Iran is unknown. This study aimed to detect Rickettsia spp. and Bartonella species in ticks in northeast Iran and conduct phylogenetic analysis on these bacteria. Ticks from the sample bank in the Research Center for Emerging and Re-emerging Diseases were included in this study. The ticks were collected in 2017 and 2018 from domestic animals (sheep, goats, cows, camels, horses, dogs, and donkeys) and rodents in Golestan, Mazandaran, and Guilan provinces. Molecular methods were used to examine the DNA extracted from these samples to detect Rickettsia spp. and Bartonella species. The study examined a total of 3999 ticks. Ixodes ricinus (46.4%), Rhipicephalus turanicus (26.3%), and Rhipicephalus sanguineus (17.1%) were the most prevalent species. Among 638 DNA pools, real-time-PCR detected Rickettsia spp. in 161 (25.2%), mostly belonging to Rh. sanguineus (48.9%) and Rh. turanicus (41.9%). Golestan Province had the highest number of positive pools (29.7%). No positive samples for Bartonella were detected in a 638 pooled samples. Eight distinct Rickettsia species were detected in 65 sequenced samples, the majority of which were R. massiliae (n = 32, 49.2%) and R. sibirica (n = 20, 30.8%). Other species included R. rhipicephali (n = 3), R. aeschlimannii (n = 5), R. helvetica (n = 5), R. asiatica (n = 4), R. monacensis (n = 6), and R. raoultii (n = 1). The research findings may provide helpful information about tick-borne Rickettsiae in Iran and help to clarify the role of these arthropods in maintaining these agents. Rickettsia species were found to be circulating in three Northern provinces; thus, it is recommended that this disease be considered in the differential diagnosis of febrile diseases caused by tick bites and febrile diseases with skin rashes such as Crimean–Congo hemorrhagic fever (CCHF). [ABSTRACT FROM AUTHOR]

Repellents serve an important role in bite protection. Tick repellents largely rely on biomechanisms that induce responses with direct contact, but synthetic pyrethroids used as spatial repellents against insects have received recent attention for potential use in tick protection systems. An in vitro vertical climb assay was designed to assess spatial repellency against Dermacentor variabilis, Amblyomma americanum, and Ixodes scapularis adult, female ticks. Climbing behavior was assessed with and without the presence of two spatial repellents, transfluthrin and metofluthrin. Repellency parameters were defined to simulate the natural questing behavior of ambushing ticks, including measures of detachment, pseudo-questing duration, climbing deterrence, and activity. Significant effects were observed within each parameter. D. variabilis showed the greatest general susceptibility to each repellent, followed by A. americanum, and I. scapularis. The most important and integrative measure of repellency was climbing deterrence–a measure of the spatial repellent's ability to disrupt a tick's natural propensity to climb. Transfluthrin deterred 75% of D. variabilis, 67% of A. americanum, and 50% of I. scapularis. Metofluthrin was slightly more effective, deterring 81% of D. variabilis, 73% of A. americanum, and 72% of I. scapularis. The present study poses a novel paradigm for repellency and reports a preliminary assessment of spatial repellent effect on tick behavior. Further research will assess spatial repellency in a more natural setting, scale exposure conditions, and incorporate host cues. [ABSTRACT FROM AUTHOR]

The distribution, abundance and seasonal activity of vector species, such as ticks and mosquitoes, are key determinants of vector-borne disease risk, and are strongly influenced by abiotic and habitat conditions. Despite the numerous species of tick vectors in the heavily populated North American West Coast, all but Ixodes pacificus, the primary vector of the Lyme disease spirochete, is poorly characterized with regard to seasonal activity patterns and fine scale drivers of distribution and abundance, particularly in heavily populated regions of southern California. This lack of knowledge inhibits both scientific understanding and public health efforts to minimize vector exposure and risk of pathogen transmission to humans. Here we address this gap by characterizing the abiotic and habitat drivers of the distribution, abundance, and diversity of the vector tick community using fine scale temporal surveys over two seasons (2014 and 2015) across coastal and inland regions of Santa Barbara County, CA. We also characterize patterns of seasonal activity of the more common vector species to understand seasonality in risk of vector exposure, and specifically focus on human encounter risk using standardized tick drags as our method of collection. Leveraging plot-level habitat and abiotic variables in partial least squares regression analysis, we find the seven different vector species collected in this study have divergent drivers of activity and abundance. For example, I. pacificus is strongly associated with dense forest habitats and cool and moist microclimates, while Dermacentor occidentalis and Dermacentor variabilis, competent vectors of Rocky Mountain Spotted Fever, were found to be more tolerant of higher average temperatures and more open habitats. These results suggest that I. pacificus may be expected to experience reductions in geographic distribution and seasonal activity under projected land cover and climate change in coastal southern California, while D. occidentalis may experience more limited effects. We discuss implications for changing tick-borne disease risk associated with pathogens transmitted by Ixodes as well as Dermacentor species ticks in the western US, and contrast these predictions with eastern North America.

Ticks are the most important vectors of zoonotic disease-causing pathogens in North America and Europe. Many tick species are expanding their geographic range. Although correlational evidence suggests that climate change is driving the range expansion of ticks, experimental evidence is necessary to develop a mechanistic understanding of ticks' response to a range of climatic conditions. Previous experiments used simulated microclimates, but these protocols require hazardous salts or expensive laboratory equipment to manipulate humidity. We developed a novel, safe, stable, convenient, and economical method to isolate individual ticks and manipulate their microclimates. The protocol involves placing individual ticks in plastic tubes, and placing six tubes along with a commercial two-way humidity control pack in an airtight container. We successfully used this method to investigate how humidity affects survival and host-seeking (questing) behavior of three tick species: the lone star tick (Amblyomma americanum), American dog tick (Dermacentor variabilis), and black-legged tick (Ixodes scapularis). We placed 72 adult females of each species individually into plastic tubes and separated them into three experimental relative humidity (RH) treatments representing distinct climates: 32% RH, 58% RH, and 84% RH. We assessed the survival and questing behavior of each tick for 30 days. In all three species, survivorship significantly declined in drier conditions. Questing height was negatively associated with RH in Amblyomma, positively associated with RH in Dermacentor, and not associated with RH in Ixodes. The frequency of questing behavior increased significantly with drier conditions for Dermacentor but not for Amblyomma or Ixodes. This report demonstrates an effective method for assessing the viability and host-seeking behavior of tick vectors of zoonotic diseases under different climatic conditions. [ABSTRACT FROM AUTHOR]

The tropical lineage within the Rhipicephalus sanguineus species complex is cause for growing concern in the U.S. based on its prominent role in creating and perpetuating multiple recently identified outbreaks of Rocky Mountain spotted fever in the southwestern United States and northern Mexico. This lineage is undergoing a northward range expansion in the United States, necessitating the need for enhanced surveillance for Rh. sanguineus. To inform more focused surveillance efforts we use species distribution models (SDMs) to predict current (2015–2019) and future (2021–2040) habitat for the tropical lineage. Models using the MaxEnt algorithm were informed using geolocations of ticks genetically confirmed to be of the tropical lineage, for which data on 23 climatic and ecological variables were extracted. Models predicted that suitability was optimal where temperatures are relatively warm and stable, and there is minimal precipitation. This translated into habitat being predicted along much of the coast of southern states including California, Texas, Louisiana, and Florida. Although the endophilic nature of tropical Rh. sanguineus somewhat violates the assumptions of SDMs, our models correctly predicted known locations of this tick and provide a starting point for increased surveillance efforts. Furthermore, we highlight the importance of using molecular methods to distinguish between ticks in the Rh. sanguineus species complex. [ABSTRACT FROM AUTHOR]

Ticks and tick-borne diseases cause substantial economic losses to the livestock industry in sub-Saharan Africa. Mazao Tickoff is a novel bioacaricide developed for tick control and is based on the entomopathogenic fungus Metarhizium anisopliae sensu lato (s.l.) isolate ICIPE 7. To date, no randomized controlled study has been undertaken to demonstrate the efficacy of this bioacaricide in reducing natural tick infestation on cattle. To this end, this field trial is designed to evaluate the anti-tick efficacy of Mazao Tickoff on cattle in coastal Kenya compared to a standard chemical tick control protocol. In this prospective, multi-center randomized controlled trial, eligible herds will be randomized by the herd size to the intervention arm in a 1:1:1 ratio to either Triatix® (active ingredient: amitraz); Mazao Tickoff (active ingredient: M. anisopliae ICIPE 7); or placebo (excipients of the Mazao Tickoff), with a total enrollment target of 1,077 cattle. Treatments will be dispensed on Day 0 (defined individually as the day each animal receives the first treatment) and thereafter every two weeks until Day 182. Ticks will be counted on every animal in each herd (herds to be included have at least one animal bearing at least one tick on Day 0), and thereafter on bi-weekly intervals until Day 182. The primary efficacy assessments of Mazao Tickoff will be based on the mean percentage reduction in tick counts at each post-treatment follow-up visit compared to the placebo group and the Triatix® arm. Further, the effect of Mazao Tickoff on the prevalence of common cattle pathogens, Anaplasma marginale and Theileria parva, will be determined by assessing incidence and seroprevalence at four different time points. This protocol describes the first rigorous evaluation of the efficacy of Mazao Tickoff and its potential as a viable alternative non-chemical acaricide tool for tick control in Kenya and elsewhere. [ABSTRACT FROM AUTHOR]

Ixodes ricinus, the most common species of tick in Europe, is known to transmit major pathogens to animals and humans such as Babesia spp. or Borrelia spp. Its abundance and distribution have been steadily increasing in Europe during recent decades, due to global environmental changes. Indeed, as ticks spend most of their life in the environment, their activity and life cycle are highly dependent on environmental conditions and therefore, on climate or habitat changes. Simultaneously, wild ungulates have expanded their range and increased dramatically in abundance worldwide, in particular roe deer (Capreolus capreolus), allowing tick populations to grow and spread. Currently, tick infestation on newborn wild ungulates is poorly documented. However, newborn ungulates are considered more sensitive to tick bites and pathogen transmission because of their immature immune systems. Thus, improving knowledge about the factors influencing tick infestation on newborns is essential to better understand their health risks. This study was conducted at Trois-Fontaines Forest, Champagne-Ardenne, France (1992–2018). Based on a long-term monitoring of roe deer fawns, we used a novel Bayesian model of the infestation of fawns to identify which biotic or abiotic factors were likely to modify the level of infestation by ticks of 965 fawns over time. We show that tick burden increased faster during the first days of life of the fawns and became constant when fawns were five days old and more, which could be explained by the depletion of questing ticks or the turnover of ticks feeding on fawns. Moreover, despite the known positive influence of humidity on tick activity, the tick burdens were weakly related to this parameter. Our results demonstrate that tick infestation was highly variable among years, particularly between 2000–2009. We hypothesize that this results from a modification of habitat caused by Hurricane Lothar. [ABSTRACT FROM AUTHOR]

Objective: This study examined the knowledge, attitudes, and practices (KAP) of an at-risk population living in Niigata prefecture regarding tick-borne diseases (TBDs) and preventive strategies. Methods: A cross-sectional questionnaire-based study was conducted to assess the KAP of the community. Results: In total, 186 responses were received. Among the respondents, 130 (69·9%) were men, and the mean age was 51.1 (14·3). Nine (4·8%) respondents reported having experienced tick bites. Of the respondents, 44 (23.7%) knew about both scrub typhus and severe fever with thrombocytopenia syndrome, while 156 (83·9%) and 71 (38·2%) recognized limiting skin exposure and use of insect repellents as preventive measures, respectively. The attitudes towards TBDs: being worried about tick bites (p = 0·018) and interested in preventing TBDs (p = 0·001), were significantly higher among women than men. About 75% of the respondents reported taking preventive measures against tick bites, and limiting skin exposure was the most frequently applied method (69·9%). Insect repellents were used by 58 (31·2%) respondents. Age (p = 0·049), being worried about tick bites (p = 0·046), and knowledge of ticks score (p = 0·024) were the significant independent predictors of practicing countermeasures. Conclusion: We identified gaps in knowledge and practices regarding TBDs. Public health interventions should be implemented to improve public awareness of TBDs. [ABSTRACT FROM AUTHOR]

Ticks are one of the most important vectors of several pathogens affecting humans and animals. In addition to pathogens, ticks carry diverse microbiota of symbiotic and commensal microorganisms. In this study, we have investigated the first Tunisian insight into the microbial composition of the most dominant Hyalomma species infesting Tunisian cattle and explored the relative contribution of tick sex, life stage, and species to the diversity, richness and bacterial species of tick microbiome. In this regard, next generation sequencing for the 16S rRNA (V3-V4 region) of tick bacterial microbiota and metagenomic analysis were established. The analysis of the bacterial diversity reveals that H. marginatum and H. excavatum have greater diversity than H. scupense. Furthermore, microbial diversity and composition vary according to the tick's life stage and sex in the specific case of H. scupense. The endosymbionts Francisella, Midichloria mitochondrii, and Rickettsia were shown to be the most prevalent in Hyalomma spp. Rickettsia, Francisella, Ehrlichia, and Erwinia are the most common zoonotic bacteria found in Hyalomma ticks. Accordingly, Hyalomma ticks could represent potential vectors for these zoonotic bacterial agents. [ABSTRACT FROM AUTHOR]

Models can be applied to extrapolate consequences of climate change for complex ecological systems in the future. The acknowledged systems' behaviour at present is projected into the future considering climate projection data. Such an approach can be used to addresses the future activity and density of the castor bean tick Ixodes ricinus, the most widespread tick species in Europe. It is an important vector of pathogens causing Lyme borreliosis and tick-borne encephalitis. The population dynamics depend on several biotic and abiotic factors. Such complexity makes it difficult to predict the future dynamics and density of I. ricinus and associated health risk for humans. The objective of this study is to force ecological models with high-resolution climate projection data to extrapolate I. ricinus tick density and activity patterns into the future. We used climate projection data of temperature, precipitation, and relative humidity for the period 1971–2099 from 15 different climate models. Tick activity was investigated using a climate-driven cohort-based population model. We simulated the seasonal population dynamics using climate data between 1971 and 2099 and observed weather data since 1949 at a specific location in southern Germany. We evaluated derived quantities of local tick ecology, e.g. the maximum questing activity of the nymphal stage. Furthermore, we predicted spatial density changes by extrapolating a German-wide tick density model. We compared the tick density of the reference period (1971–2000) with the counter-factual densities under the near-term scenario (2012–2041), mid-term scenario (2050–2079) and long-term scenario (2070–2099). We found that the nymphal questing peak would shift towards early seasons of the year. Also, we found high spatial heterogeneity across Germany, with predicted hotspots of up to 2,000 nymphs per 100 m2 and coldspots with constant density. As our results suggest extreme changes in tick behaviour and density, we discuss why caution is needed when extrapolating climate data-driven models into the distant future when data on future climate drive the model projection. [ABSTRACT FROM AUTHOR]

Ixodes scapularis ticks are expanding their range in parts of northeastern North America, bringing with them pathogens of public health concern. While rodents like the white-footed mouse, Peromyscus leucopus, are considered the primary reservoir of many emerging tick-borne pathogens, the contribution of birds, as alternative hosts and reservoirs, to local transmission cycles has not yet been firmly established. From 2016 to 2018, we collected host-seeking ticks and examined rodent and bird hosts for ticks at 48 sites in a park where blacklegged ticks are established in Quebec, Canada, in order to characterize the distribution of pathogens in ticks and mammalian and avian hosts. We found nearly one third of captured birds (n = 849) and 70% of small mammals (n = 694) were infested with I. scapularis. Five bird and three mammal species transmitted Borrelia burgdorferi to feeding larvae (n larvae tested = 2257) and we estimated that about one fifth of the B. burgdorferi-infected questing nymphs in the park acquired their infection from birds, the remaining being attributable to mice. Ground-foraging bird species were more parasitized than other birds, and species that inhabited open habitat were more frequently infested and were more likely to transmit B. burgdorferi to larval ticks feeding upon them. Female birds were more likely to transmit infection than males, without age differentiation, whereas in mice, adult males were more likely to transmit infection than juveniles and females. We also detected Borrelia miyamotoi in larvae collected from birds, and Anaplasma phagocytophilum from a larva collected from a white-footed mouse. This study highlights the importance of characterising the reservoir potential of alternative reservoir hosts and to quantify their contribution to transmission dynamics in different species assemblages. This information is key to identifying the most effective host-targeted risk mitigation actions. [ABSTRACT FROM AUTHOR]

The incidence of Lyme disease is increasing in Québec and is closely linked to the distribution of Ixodes scapularis ticks. A time-to-establishment model developed in 2012 by Leighton and colleagues predicted the year of tick population establishment for each municipality in eastern Canada. To validate if this model correctly predicted tick distribution in Québec, predicted tick establishment was compared to field data from active tick surveillance (2010–2018) using two criteria: i) the detection of at least one tick and ii) the detection of the three questing stages of the tick. The speed of tick establishment and the increase in the exposed human population by 2100 were predicted with the time-to-establishment model. Field observations were consistent with model predictions. Ticks were detected on average 3 years after the predicted year. The probability of tick detection is significantly higher after the predicted year than before (61% vs 27% of collections). The trend was similar for the detection of three tick stages (16% vs 9% of collections). The average speed of tick range expansion was estimated by the model to be 18 km/year in Québec, with 90% of the human population exposed by 2027. The validation of the time-to-establishment model using field data confirmed that it could be used to project I. scapularis range expansion in Québec, and consequently the increase in Lyme disease risk over the coming decades. This will help public health authorities anticipate and adapt preventive measures, especially in areas not yet affected by Lyme disease. [ABSTRACT FROM AUTHOR]

Borrelia burgdorferi and Borrelia miyamotoi are tick-vectored zoonotic pathogens maintained in wildlife species. Tick populations are establishing in new areas globally in response to climate change and other factors. New Brunswick is a Canadian maritime province at the advancing front of tick population establishment and has seen increasing numbers of ticks carrying B. burgdorferi, and more recently B. miyamotoi. Further, it is part of a region of Atlantic Canada with wildlife species composition differing from much of continental North America and little information exists as to the presence and frequency of infection of Borrelia spp. in wildlife in this region. We used a citizen science approach to collect a wide range of animals including migratory birds, medium-sized mammals, and small mammals. In total we tested 339 animals representing 20 species for the presence of B. burgdorferi and B. miyamotoi. We have developed new nested PCR primers and a protocol with excellent specificity for detecting both of these Borrelia species, both single and double infections, in tissues and organs of various wildlife species. The positive animals were primarily small non-migratory mammals, approximately twice as many were infected with B. burgdorferi than B. miyamotoi and one animal was found infected with both. In addition to established reservoir species, the jumping mouse (Napaeozapus insignis) was found frequently infected; this species had the highest infection prevalence for both B. burgdorferi and B. miyamotoi and has not previously been identified as an important carrier for either Borrelia species. Comprehensive testing of tissues found that all instances of B. burgdorferi infection were limited to one tissue within the host, whereas two of the five B. miyamotoi infections were diffuse and found in multiple systems. In the one coinfected specimen, two fetuses were also recovered and found infected with B. miyamotoi. This presumptive transplacental transmission suggests that vertical transmission in mammals is possible. This finding implies that B. miyamotoi could rapidly spread into wildlife populations, as well as having potential human health implications. [ABSTRACT FROM AUTHOR]

Based on chromosome sequences, the human pathogen Borrelia miyamotoi phylogenetically clusters with species that cause relapsing fever. But atypically for relapsing fever agents, B. miyamotoi is transmitted not by soft ticks but by hard ticks, which also are vectors of Lyme disease Borrelia species. To further assess the relationships of B. miyamotoi to species that cause relapsing fever, I investigated extrachromosomal sequences of a North American strain with specific attention on plasmid-borne vsp and vlp genes, which are the underpinnings of antigenic variation during relapsing fever. For a hybrid approach to achieve assemblies that spanned more than one of the paralogous vsp and vlp genes, a database of short-reads from next-generation sequencing was supplemented with long-reads obtained with real-time DNA sequencing from single polymerase molecules. This yielded three contigs of 31, 16, and 11 kb, which each contained multiple and diverse sequences that were homologous to vsp and vlp genes of the relapsing fever agent B. hermsii. Two plasmid fragments had coding sequences for plasmid partition proteins that differed from each other from paralogous proteins for the megaplasmid and a small plasmid of B. miyamotoi. One of 4 vsp genes, vsp1, was present at two loci, one of which was downstream of a candiate prokaryotic promoter. A limited RNA-seq analysis of a population growing in the blood of mice indicated that of the 4 different vsp genes vsp1 was the one that was expressed. The findings indicate that B. miyamotoi has at least four types of plasmids, two or more of which bear vsp and vlp gene sequences that are as numerous and diverse as those of relapsing fever Borrelia. The database and insights from these findings provide a foundation for further investigations of the immune responses to this pathogen and of the capability of B. miyamotoi for antigenic variation.

Adaptation is key for survival as vector-borne pathogens transmit between the arthropod and vertebrate, and temperature change is an environmental signal inducing alterations in gene expression of tick-borne spirochetes. While plasmids are often associated with adaptation, complex genomes of relapsing fever spirochetes have hindered progress in understanding the mechanisms of vector colonization and transmission. We utilized recent advances in genome sequencing to generate the most complete version of the Borrelia turicatae 150 kb linear megaplasmid (lp150). Additionally, a transcriptional analysis of open reading frames (ORFs) in lp150 was conducted and identified regions that were up-regulated during in vitro cultivation at tick-like growth temperatures (22°C), relative to bacteria grown at 35°C and infected murine blood. Evaluation of the 3' end of lp150 identified a cluster of ORFs that code for putative surface lipoproteins. With a microbe's surface proteome serving important roles in pathogenesis, we confirmed the ORFs expression in vitro and in the tick compared to spirochetes infecting murine blood. Transcriptional evaluation of lp150 indicates the plasmid likely has essential roles in vector colonization and/or initiating mammalian infection. These results also provide a much needed transcriptional framework to delineate the molecular mechanisms utilized by relapsing fever spirochetes during their enzootic cycle.

Preventing bites from undetected ticks through bathing practices would benefit public health, but the effects of these practices have been researched minimally. We immersed nymphal and adult hard ticks of species common in the eastern United States in tap water, using temperatures and durations that are realistic for human hot bathing. The effect of (a) different skin-equivalent surfaces (silicone and pig skin), and (b) water temperature was tested on Amblyomma americanum, Dermacentor variabilis and Ixodes scapularis nymphs. Overall, the type of surface had a much larger effect on the nymphs' tendency to stay in contact with the surface than water temperature did. Most nymphs that separated from the surface did so within the first 10 s of immersion, with the majority losing contact due to the formation of an air bubble between their ventral side and the test surface. In addition, adult Ixodes scapularis were tested for the effect of immersion time, temperature, and soap on tick responsiveness. Some individual adults moved abnormally or stopped moving as a result of longer or hotter immersion, but soap had little effect on responsiveness. Taken together, our results suggest that the surface plays a role in ticks' tendency to stay in contact; the use of different bath additives warrants further research. While water temperature did not have a significant short-term effect on tick separation, ticks that have not attached by their mouth parts may be rendered unresponsive and eventually lose contact with a person's skin in a hot bath. It should be noted that our research did not consider potential temperature effects on the pathogens themselves, as previous research suggests that some tickborne pathogens may become less hazardous even if the tick harboring them survives hot-water exposures and later bites the bather after remaining undetected. [ABSTRACT FROM AUTHOR]

A recent research study on prevalence of tick-borne pathogens in Burundi reported high prevalence and endemicity of Theileria parva, Anaplasma marginale and Babesia bigemina infections in cattle. Detailed information about tick species infesting animals, their distribution and genetic diversity in Burundi is outdated and limited. This study therefore assessed the prevalence and genetic diversity of tick species infesting cattle across agroecological zones (AEZs) in Burundi. A cross-sectional study on the occurrence of tick species was conducted in 24 districts of Burundi between October and December 2017. Differential identification and characterization of ticks collected was conducted using tick morphological keys and molecular tools (cox1 and 12S rRNA gene). Chi-square test was used to test for association between agroecological zones and the prevalence of tick species. Phylogenetic relationships were inferred using bayesian and maximum likelihood algorithms. A total of 483 ticks were collected from the five AEZs sampled. Six tick species comprising of Rhipicephalus appendiculatus, R. sanguineus, R. evertsi evertsi, R. microplus, R. decoloratus and Amblyomma variegatum were observed. Rhipicephalus appendiculatus were the most prevalent ticks (~45%). A total of 138 specimens (28%) were found to be Rhipicephalus microplus, suggesting an emerging threat for cattle farmers. Twelve R. appendiculatus cox1 haplotypes were obtained from 106 specimens that were sequenced. Two cox1 haplotypes of R. microplus which clustered into previously reported Clade A were observed. Rhipicephalus sanguineus and R. evertsi evertsi ticks, the vectors of numerous zoonotic pathogens, were collected from cattle, which constitute a high risk for public health. These findings reveal an overlapping distribution of tick vectors in Burundi. The design of ticks and tick-borne diseases control strategies should consider the distribution of different vectors across the AEZs particularly the presence of the highly invasive R. microplus tick in Burundi and the potential risk of introducing the pathogenic Babesia bovis. [ABSTRACT FROM AUTHOR]

Ticks and tick-borne diseases represent a growing public health threat in North America and Europe. The number of ticks, their geographical distribution, and the incidence of tick-borne diseases, like Lyme disease, are all on the rise. Accurate, real-time tick-image identification through a smartphone app or similar platform could help mitigate this threat by informing users of the risks associated with encountered ticks and by providing researchers and public health agencies with additional data on tick activity and geographic range. Here we outline the requirements for such a system, present a model that meets those requirements, and discuss remaining challenges and frontiers in automated tick identification. We compiled a user-generated dataset of more than 12,000 images of the three most common tick species found on humans in the U.S.: Amblyomma americanum, Dermacentor variabilis, and Ixodes scapularis. We used image augmentation to further increase the size of our dataset to more than 90,000 images. Here we report the development and validation of a convolutional neural network which we call "TickIDNet," that scores an 87.8% identification accuracy across all three species, outperforming the accuracy of identifications done by a member of the general public or healthcare professionals. However, the model fails to match the performance of experts with formal entomological training. We find that image quality, particularly the size of the tick in the image (measured in pixels), plays a significant role in the network's ability to correctly identify an image: images where the tick is small are less likely to be correctly identified because of the small object detection problem in deep learning. TickIDNet's performance can be increased by using confidence thresholds to introduce an "unsure" class and building image submission pipelines that encourage better quality photos. Our findings suggest that deep learning represents a promising frontier for tick identification that should be further explored and deployed as part of the toolkit for addressing the public health consequences of tick-borne diseases. [ABSTRACT FROM AUTHOR]

Unexpected questing activity of ticks was noted during the winter months of January and February in the Central Midwestern states of Kansas, Missouri, Oklahoma, and Arkansas. From nine geographically distinct locations, four species of ticks were collected using the flagging method, of which the lone star tick, Amblyomma americanum, was most abundant, followed by the American dog tick, Dermacentor variabilis, the Gulf coast tick, Amblyomma maculatum, and the Black legged tick, Ixodes scapularis. More A. americanum nymphs were caught questing than male or female adults. The winter activity of these medically important ticks in this region poses concern for public health and offers an insight into future tick activity in light of ongoing climate change. More studies on the seasonality of these tick species, and how different climate parameters affect their seasonal activity in this region are warranted and would be expected to benefit for both human and veterinary medicine. [ABSTRACT FROM AUTHOR]

In this study, we investigated the presence of tick-borne pathogens in ticks removed from tick-bitten humans in the southwestern provinces of the Republic of Korea (ROK). We identified 33 ticks from three tick species, namely Amblyomma testudinarium (60.6%), Haemaphysalis longicornis (27.3%), and Ixodes nipponensis (12.1%) in order of occurrence via morphology and 16S rDNA-targeting polymerase chain reaction (PCR). Tick-borne pathogens were detected in 16 ticks using pathogen-specific PCR. From the results, 12 ticks (36.4%) tested positive for spotted fever group (SFG) Rickettsia: Rickettsia monacensis (1/12), R. tamurae (8/12), and Candidatus Rickettsia jingxinensis (3/12). Three ticks (9.1%) were positive for Anaplasma phagocytophilum. In addition, three ticks (9.1%) tested positive for Babesia gibsoni (1/3) and B. microti (2/3). In conclusion, we identified three tick species; the most common species was A. testudinarium, followed by H. longicornis and I. nipponensis. SFG Rickettsia, A. phagocytophilum, and Babesia spp. were the most frequently detected pathogens in ticks removed from tick-bitten humans. To our knowledge, this is the first report of R. tamurae and Ca. R. jingxinensis detection in Korea. The present results will contribute to the understanding of tick-borne infections in animals and humans in the ROK. [ABSTRACT FROM AUTHOR]

Between March 2014 and February 2017, host-seeking ticks were collected during the late spring and summer months seasonally, and as well as continually through all seasons from several sites in a periurban environment in Pittsburg, Kansas, located in the Central Midwestern United States. All three post-emergent life-stages of Amblyomma americanum, and the adults of three other ticks viz. Dermacentor variabilis, A. maculatum, and Ixodes scapularis were collected using the flagging method, and were taxonomically identified using morphological and molecular methods. A total of 15946 ticks were collected from these sites. A vast majority of the ticks collected over the three-year study period was A. americanum (79.01%). The three other species collected included D. variabilis (13.10%), A. maculatum (7.15%), and Ixodes scapularis (0.73%). More female ticks of each species were collected throughout the study period from all sites, and a unimodal activity period was noted for all four species. The diversity, composition, and phenology of these medically significant tick species are discussed. [ABSTRACT FROM AUTHOR]

Background: Tick-borne encephalitis (TBE) is the most important tick-borne viral disease in Eurasia and causes disease in humans and in a number of animals, among them dogs and horses. There is still no good correlation between tick numbers, weather conditions and human cases. There is the hypothesis that co-feeding due to simultaneous occurrence of larvae and nymphs may be a factor for the increased transmission of the virus in nature and for human disease. Based on long-term data from a natural TBEV focus, phylogenetic results and meteorological data we sought to challenge this hypothesis. Methods: Ticks from an identified TBE natural focus were sampled monthly from 04/2009 to 12/2018. Ticks were identified and pooled. Pools were tested by RT-qPCR. Positive pools were confirmed by virus isolation and/or sequencing of additional genes (E gene, NS2 gene). Temperature data such as the decadal (10-day) mean daily maximum air temperature (DMDMAT) were obtained from a nearby weather station and statistical correlations between tick occurrence and minimal infection rates (MIR) were calculated. Results: In the study period from 04/2009 to 12/2018 a total of 15,530 ticks (2,226 females, 2,268 males, 11,036 nymphs) were collected. The overall MIR in nymphs over the whole period was 77/15,530 (0.49%), ranging from 0.09% (2009) to 1.36% (2015). The overall MIR of female ticks was 0.76% (17/2,226 ticks), range 0.14% (2013) to 3.59% (2016). The overall MIR of males was 0.57% (13/2,268 ticks), range from 0.26% (2009) to 0.97% (2015). The number of nymphs was statistically associated with a later start of spring/vegetation period, indicated by the onset of forsythia flowering. Conclusion: There was no particular correlation between DMDMAT dynamics in spring and/or autumn and the MIR of nymphs or adult ticks detected. However, there was a positive correlation between the number of nymphs and the number of reported human TBE cases in the following months, but not in the following year. The hypothesis of the importance of co-feeding of larvae and nymphs for the maintenance of transmission cycle of TBEV in nature is not supported by our findings. [ABSTRACT FROM AUTHOR]

In the twenty-first century, ticks and tick-borne diseases have expanded their ranges and impact across the US. With this spread, it has become vital to monitor vector and disease distributions, as these shifts have public health implications. Typically, tick-borne disease surveillance (e.g., Lyme disease) is passive and relies on case reports, while disease risk is calculated using active surveillance, where researchers collect ticks from the environment. Case reports provide the basis for estimating the number of cases; however, they provide minimal information on vector population or pathogen dynamics. Active surveillance monitors ticks and sylvatic pathogens at local scales, but it is resource-intensive. As a result, data are often sparse and aggregated across time and space to increase statistical power to model or identify range changes. Engaging public participation in surveillance efforts allows spatially and temporally diverse samples to be collected with minimal effort. These citizen-driven tick collections have the potential to provide a powerful tool for tracking vector and pathogen changes. We used MaxEnt species distribution models to predict the current and future distribution of Ixodes pacificus across the Western US through the use of a nationwide citizen science tick collection program. Here, we present niche models produced through citizen science tick collections over two years. Despite obvious limitations with citizen science collections, the models are consistent with previously-predicted species ranges in California that utilized more than thirty years of traditional surveillance data. Additionally, citizen science allows for an expanded understanding of I. pacificus distribution in Oregon and Washington. With the potential for rapid environmental changes instigated by a burgeoning human population and rapid climate change, the development of tools, concepts, and methodologies that provide rapid, current, and accurate assessment of important ecological qualities will be invaluable for monitoring and predicting disease across time and space. [ABSTRACT FROM AUTHOR]

Tick-borne relapsing fever is an infectious disease caused by Borrelia species and are primarily transmitted by Ornithodoros ticks. Prior work indicated that in vitro cultivated spirochetes remain infectious to mice by needle inoculation; however, the impact of laboratory propagation on the pathogens natural life cycle has not been determined. Our current study assessed the effect of serial cultivation on the natural tick-mammalian transmission cycle. First, we evaluated genomic DNA profiles from B. turicatae grown to 30, 60, 120, and 300 generations, and these spirochetes were used to needle inoculate mice. Uninfected nymphal ticks were fed on these mice and acquisition, transstadial maintenance, and subsequent transmission after tick bite was determined. Infection frequencies in mice that were fed upon by ticks colonized with B. turicatae grown to 30, 60, and 120 generations were 100%, 100%, and 30%, respectively. Successful infection of mice by tick feeding was not detected after 120 generations. Quantifying B. turicatae in tick tissues indicated that by 300 generations they no longer colonized the vector. The results indicate that in vitro cultivation significantly affects the establishment of tick colonization and murine infection. This work provides a foundation for the identification of essential genetic elements in the tick-mammalian infectious cycle. [ABSTRACT FROM AUTHOR]

In studies assessing tick abundance, the use of live traps to capture and euthanize rodent hosts is a commonly used method to determine their burden. However, captive animals can experience debilitating or fatal capture stress as a result prior to collection. An alternative method is the use of lethal traps, but this can potentially lead to tick drop-off between the time of capture and collection. In this study, in order to determine whether subjecting animals to capture stress is inevitable, we tested the difference in sheep tick (Ixodes ricinus) larval burdens between bank voles (Myodes glareolus) captured alive and euthanized, and lethally trapped bank voles. During 2017 and 2018, 1318 bank voles were captured using live (Ugglan Special no. 2) and lethal (Rapp2 Mousetrap) traps during two consecutive years over three seasons in two locations in Norway. Voles captured alive would remain captive until euthanized, while lethally trapped voles were killed instantly upon capture. Log-linear models, accounting for overdispersion, were used to determine whether trap type was influencing observed tick burden. Bank voles captured in lethal traps carried 5.7% more larvae compared to euthanized voles captured in live traps, but this difference was not significant (p = 0.420). Males were overall captured 2.7 times more frequently than females, and the sex ratio was equal in both trap types. This study shows that the use of lethal traps to determine tick burden of rodents is sufficiently reliable, without having to subject animals to potentially lethal stress, hereby reducing some ethical concerns of animal suffering and the results thereof, without compromising accuracy. Lethal trapping is also often more economical and practical, further favoring this collection method. [ABSTRACT FROM AUTHOR]

The role of questing ticks in the epidemiology of tick-borne diseases in Kenya's Maasai Mara National Reserve (MMNR), an ecosystem with intensified human-wildlife-livestock interactions, remains poorly understood. We surveyed the diversity of questing ticks, their blood-meal hosts, and tick-borne pathogens to understand potential effects on human and livestock health. By flagging and hand-picking from vegetation in 25 localities, we collected 1,465 host-seeking ticks, mostly Rhipicephalus and Amblyomma species identified by morphology and molecular analysis. We used PCR with high-resolution melting (HRM) analysis and sequencing to identify Anaplasma, Babesia, Coxiella, Ehrlichia, Rickettsia, and Theileria pathogens and blood-meal remnants in 231 tick pools. We detected blood-meals from humans, wildebeest, and African buffalo in Rh. appendiculatus, goat in Rh. evertsi, sheep in Am. gemma, and cattle in Am. variegatum. Rickettsia africae was detected in Am. gemma (MIR = 3.10) that had fed on sheep and in Am. variegatum (MIR = 250) that had fed on cattle. We found Rickettsia spp. in Am. gemma (MIR = 9.29) and Rh. evertsi (MIR = 200), Anaplasma ovis in Rh. appendiculatus (MIR = 0.89) and Rh. evertsi (MIR = 200), Anaplasma bovis in Rh. appendiculatus (MIR = 0.89), and Theileria parva in Rh. appendiculatus (MIR = 24). No Babesia, Ehrlichia, or Coxiella pathogens were detected. Unexpectedly, species-specific Coxiella sp. endosymbionts were detected in all tick genera (174/231 pools), which may affect tick physiology and vector competence. These findings show that ticks from the MMNR are infected with zoonotic R. africae and unclassified Rickettsia spp., demonstrating risk of African tick-bite fever and other spotted-fever group rickettsioses to locals and visitors. The protozoan pathogens identified may also pose risk to livestock production. The diverse vertebrate blood-meals of questing ticks in this ecosystem including humans, wildlife, and domestic animals, may amplify transmission of tick-borne zoonoses and livestock diseases. [ABSTRACT FROM AUTHOR]

The American dog tick, Dermacentor variabilis, is a veterinary- and medically- significant tick species that is known to transmit several diseases to animal and human hosts. The spatial distribution of this species in North America is not well understood, however; and knowledge of likely changes to its future geographic distribution owing to ongoing climate change is needed for proper public health planning and messaging. Two recent studies have evaluated these topics for D. variabilis; however, less-rigorous modeling approaches in those studies may have led to erroneous predictions. We evaluated the present and future distribution of this species using a correlative maximum entropy approach, using publicly available occurrence information. Future potential distributions were predicted under two representative concentration pathway (RCP) scenarios; RCP 4.5 for low-emissions and RCP 8.5 for high-emissions. Our results indicated a broader current distribution of this species in all directions relative to its currently known extent, and dramatic potential for westward and northward expansion of suitable areas under both climate change scenarios. Implications for disease ecology and public health are discussed. [ABSTRACT FROM AUTHOR]

Rhipicephalus microplus is recognized as a tick species highly prevalent in cattle, with a wide pantropical distribution that seems to continue spreading geographically. However, its role as a biological vector has been scarcely studied in the livestock context. In this study, a 16S rRNA next-generation sequencing analysis was used to determine bacterial diversity in salivary glands and gut of R. microplus from two contrasting livestock agroecosystems in Antioquia, Colombia. Both the culture-independent approach (CI) and the culture-dependent (CD) approach were complementarily adopted in this study. A total of 341 unique OTUs were assigned, the richness showed to be higher in the Northern than in the Middle Magdalena region, and a high diversity was found at the phylum and genus levels in the samples obtained. With the CI approach, Proteobacteria, Bacteroidetes, Firmicutes, and Actinobacteria were the most common phylum of bacteria regardless of the organ, or geographic origin of the specimens analyzed. While the relative abundance of bacteria at a phylum level with the CD approach varied between analyzed samples, the data obtained suggest that a high diversity of species of bacteria occurs in R. microplus from both livestock agroecosystems. Bacterial genera such as Anaplasma, Coxiella, and Ehrlichia, recognized for their implications in tick-borne diseases, were also detected, together with endosymbionts such as Lysinibacillus, previously reported as a potential tool for biological control. This information is useful to deepen the knowledge about microbial diversity regarding the relations between endosymbionts and pathogens and could facilitate the future development of epidemiological surveillance in livestock systems. [ABSTRACT FROM AUTHOR]

Birds are important hosts for the first life stages of the Ixodes ricinus tick and they can transport their parasites over long distances. The aim of this study was to investigate the prevalence of Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum, Neoehrlichia mikurensis and Rickettsia helvetica in ticks collected from migratory birds in Norway. A total of 815 Ixodes ricinus ticks from 216 birds trapped at Lista Bird Observatory in southern Norway during spring and autumn migration in 2008 were analysed by real-time PCR. B. burgdorferi s. l. was the most prevalent pathogen, detected in 6.1% of the ticks. The prevalence of N. mikurensis, A. phagocytophilum and R. helvetica was 1.2%, 0.9% and 0.4% respectively. In addition, one sample (0.1%) was positive for B. miyamotoi. In total, 8.2% of the ticks were infected with at least one pathogen. Co-infection with B. burgdorferi s. l. and N. mikurensis or A. phagocytophilum was found in 6.0% of the infected ticks. Our results show that all the known major tick-borne bacterial pathogens in Norway are subject to transport by migratory birds, potentially allowing spread to new areas. Our study showed a surprisingly high number of samples with PCR inhibition (57%). These samples had been extracted using standard methodology (phenol-chloroform extraction). This illustrates the need for inhibition controls to determine true prevalence rates. [ABSTRACT FROM AUTHOR]

The order Piroplasmida encompasses two main families: Babesiidae and Theileriidae, containing tick-borne pathogens of veterinary and medical importance worldwide. While only three genera (Babesia, Cytauxzoon and Theileria) comprising piroplasm parasites are currently recognised, phylogenetic studies at the 18S rRNA (18S) gene suggest that these organisms represent at least ten lineages, one of which comprises the relatively unique and highly diverse Theileria spp. from Australian marsupials and ticks. As an alternative to analysing 18S sequences alone, sequencing of mitochondrial genes has proven to be useful for the elucidation of evolutionary relationships amongst some groups of piroplasms. This research aimed to characterise piroplasms from Australian native mammals and ticks using multiple genetic markers (18S, cytochrome c, oxidase subunit III (cox3) and cytochrome B (cytB)) and microscopy. For this, nearly complete piroplasm-18S sequences were obtained from 32 animals belonging to six marsupial species: eastern bettong (Bettongia gaimardi), eastern quoll (Dasyurus viverrinus), eastern grey kangaroo (Macropus giganteus), swamp wallaby (Wallabia bicolor), quokka (Setonix brachyurus) and Gilbert's potoroo (Potorous gilbertii). The organisms detected represented eight novel Theileria genotypes, which formed five sub-clades within the main marsupial clade containing previously reported Australian marsupial and tick-derived Theileria spp. A selection of both novel and previously described Australian piroplasms at the 18S were also successfully characterised, for the first time, at the cox3 and cytB loci, and corroborated the position of Australian native theilerias in a separate, well-supported clade. Analyses of the cox3 and cytB genes also aided in the taxonomic resolution within the clade of Australian Piroplasmida. Importantly, microscopy and molecular analysis at multiple loci led to the discovery of a unique piroplasm species that clustered with the Australian marsupial theilerias, for which we propose the name Theileria lupei n. sp. [ABSTRACT FROM AUTHOR]