Molecular Analysis of Tick-Borne Bacterial Pathogens from Ticks Infesting Animal Hosts in Kyrgyzstan, 2021.

This study investigated the prevalence of Anaplasma and Ehrlichia in 494 engorged ticks collected from various animal hosts, including cattle, horses, sheep, chickens, dogs, and cats, in six regions of northern Kyrgyzstan. Ten tick species, belonging to two families and six genera, were identified based on CO1, 16S rRNA, and ITS2 genes: Argas persicus (26.5%), Haemaphysalis punctata (18.0%), Dermacentor spp. (16.0%), Rhipicephalus annulatus (11.8%), R. turanicus (10.9%), D. marginatus (7.7%), Hyalomma scupense (4.5%), Hy. marginatum (3.8%), R. sangineus complex (0.6%), and Ornithodoros lahorensis (0.2%). PCR analysis revealed a 15.0% (74/494) overall infection rate of Anaplasma and Ehrlichia. Anaplasma species were found in six tick species and were identified as A. bovis (n = 44), Anaplasma spp. (n = 20), A. ovis (n = 5), and A. capra (n = 2). Ehrlichia species were found only in H. punctata (n = 5) and identified as E. chaffeensis (n = 1) and Ehrlichia spp. (n = 4). Additionally, two H. punctata were co-infected with Anaplasma and Ehrlichia. This is the first study to investigate tick-borne bacterial pathogens in ticks collected from animal hosts in Kyrgyzstan. Our findings contribute to a better understanding of the epidemiology and emergence of tick-borne infections in Kyrgyzstan.

Xinyang flavivirus, from Haemaphysalis flava ticks in Henan Province, China, defines a basal, likely tick-only Orthoflavivirus clade.

Tick-borne orthoflaviviruses (TBFs) are classified into three conventional groups based on genetics and ecology: mammalian, seabird and probable-TBF group. Recently, a fourth basal group has been identified in Rhipicephalus ticks from Africa: Mpulungu flavivirus (MPFV) in Zambia and Ngoye virus (NGOV) in Senegal. Despite attempts, isolating these viruses in vertebrate and invertebrate cell lines or intracerebral injection of newborn mice with virus-containing homogenates has remained unsuccessful. In this study, we report the discovery of Xinyang flavivirus (XiFV) in Haemaphysalis flava ticks from Xìnyáng, Henan Province, China. Phylogenetic analysis shows that XiFV was most closely related to MPFV and NGOV, marking the first identification of this tick orthoflavivirus group in Asia. We developed a reverse transcriptase quantitative PCR assay to screen wild-collected ticks and egg clutches, with absolute infection rates of 20.75 % in adult females and 15.19 % in egg clutches, suggesting that XiFV could be potentially spread through transovarial transmission. To examine potential host range, dinucleotide composition analyses revealed that XiFV, MPFV and NGOV share a closer composition to classical insect-specific orthoflaviviruses than to vertebrate-infecting TBFs, suggesting that XiFV could be a tick-only orthoflavivirus. Additionally, both XiFV and MPFV lack a furin cleavage site in the prM protein, unlike other TBFs, suggesting these viruses might exist towards a biased immature particle state. To examine this, chimeric Binjari virus with XIFV-prME (bXiFV) was generated, purified and analysed by SDS-PAGE and negative-stain transmission electron microscopy, suggesting prototypical orthoflavivirus size (~50 nm) and bias towards uncleaved prM. In silico structural analyses of the 3'-untranslated regions show that XiFV forms up to five pseudo-knot-containing stem-loops and a prototypical orthoflavivirus dumbbell element, suggesting the potential for multiple exoribonuclease-resistant RNA structures.

Identification of Anaplasma marginale adhesins for entry into Dermacentor andersoni tick cells using phage display.

Anaplasma marginale is an obligate, intracellular, tick-borne bacterial pathogen that causes bovine anaplasmosis, an often severe, production-limiting disease of cattle found worldwide. Methods to control this disease are lacking, in large part due to major knowledge gaps in our understanding of the molecular underpinnings of basic host-pathogen interactions. For example, the surface proteins that serve as adhesins and, thus, likely play a role in pathogen entry into tick cells are largely unknown. To address this knowledge gap, we developed a phage display library and screened 66 A. marginale proteins for their ability to adhere to Dermacentor andersoni tick cells. From this screen, 17 candidate adhesins were identified, including OmpA and multiple members of the Msp1 family, including Msp1b, Mlp3, and Mlp4. We then measured the transcript of ompA and all members of the msp1 gene family through time, and determined that msp1b, mlp2, and mlp4 have increased transcript during tick cell infection, suggesting a possible role in host cell binding or entry. Finally, Msp1a, Msp1b, Mlp3, and OmpA were expressed as recombinant protein. When added to cultured tick cells prior to A. marginale infection, all proteins except the C-terminus of Msp1a reduced A. marginale entry by 2.2- to 4.7-fold. Except OmpA, these adhesins lack orthologs in related pathogens of humans and animals, including Anaplasma phagocytophilum and the Ehrlichia spp., thus limiting their utility in a universal tick transmission-blocking vaccine. However, this work greatly advances efforts toward developing methods to control bovine anaplasmosis and, thus, may help improve global food security.

Molecular detection of Borrelia theileri in cattle in Korea.

Bovine borreliosis, caused by Borrelia theileri which is transmitted via hard tick bites, is associated with mild clinical symptoms, such as fever, lethargy, hemoglobinuria, anorexia, and anemia. Borrelia theileri infects various animals, such as cattle, deer, horses, goats, sheep, and wild ruminants, in Africa, Australia, and South America. Notably, no case of B. theileri infection has been reported in Korean cattle to date. In this study, 101 blood samples were collected from a Korean indigenous cattle breed, among which 1.98% tested positive for B. theileri via nested PCR. The obtained sequences exhibited high homology with B. theileri strains identified in other regions. Phylogenetic analysis of 16S rRNA confirmed the B. theileri group affiliation; however, flagellin B sequences exhibited divergence, potentially due to regional evolutionary differences. This study provides the first molecular confirmation of B. theileri infection in Korean livestock. Further isolation and nucleotide sequence analyses are necessary to better understand the presence of B. theileri strains in cows in Korea.

Cascading impacts of overstory structure in managed forests on understory structure, microclimate conditions, and Ixodes scapularis (Acari: Ixodidae) densities.

Forest management practices designed to meet varied landowner objectives affect wildlife habitat and may interrupt the life-cycle stages of disease vectors, including the black-legged tick, Ixodes scapularis Say (Acari: Ixodidae). Ixodes scapularis transmits multiple pathogens including Borrelia burgdorferi, the causative agent of Lyme disease, which is the most common tick-borne disease in the United States. There is evidence that a range of active forest management practices (e.g., invasive plant removal, prescribed burning) can alter tick densities and pathogen transmission. However, few studies have investigated relationships between forest stand structural variables commonly manipulated by timber harvesting and tick ecology. Foresters may harvest timber to create certain forest structural conditions like the mean number of trees, or basal area, per hectare. This study used a spatially replicated experiment in a blocked design to compare forest stands with a range of overstory structures and document variations in the midstory, understory, and forest floor, as well as microclimate conditions within tick off-host habitat. Greater numbers of trees or basal area per hectare correlated with greater canopy closure but less understory cover, stabilized microclimate temperature, higher microclimate humidity, and greater I. scapularis nymph densities. A random forest model identified understory forest structure as the strongest predictor of nymph densities. There was no relationship between the number of trees or basal area per hectare and daily deer (Odocoileus virginianus Zimmermann) activity or nymphal infection prevalence. These findings provide a deeper understanding of tick-habitat associations within a forest stand and have the potential to inform forest management decisions.

Detection of Anaplasma spp. and Ehrlichia spp. in dogs from a veterinary teaching hospital in Italy: a retrospective study 2012-2020.

Anaplasma phagocytophilum, Anaplasma platys and Ehrlichia canis, responsible of diseases in dogs, are tick-borne pathogens with a proven or potential zoonotic role that have shown increasing prevalence worldwide. The aims of this retrospective study were to assess the frequency of Anaplasma spp. and Ehrlichia spp. exposure in dogs tested in a veterinary teaching hospital in Italy over a 9-year period, to compare the performance of the diagnostic tests used, to evaluate correlations with clinical data, and to genetically analyse the identified bacteria. During the study period, 1322 dogs tested by at least one of the rapid immunoenzymatic test, indirect immunofluorescent antibody test or end-point PCR assay for Anaplasmataceae detection were included. Dogs were tested if they had clinical signs or clinicopathological alteration or risk factors related to infection, and if they were potential blood-donor animals. Ninety-four of 1322 (7.1%) dogs tested positive for at least one pathogen: 53 (4.3%) for A. phagocytophilum, one (0.1%) for A. platys and 63 (4.6%) for E. canis. The number of dogs tested increased and the positivity rate progressively declined over the years. Comparison of tests showed a near-perfect agreement between serological tests and a poor agreement between PCR and indirect assays. A breed predisposition has been highlighted for A. phagocytophilum infection in hunting breed dogs and for E. canis infection in mixed breed dogs. Phylogeny confirmed potential zoonotic implications for A. phagocytophilum and showed no correlation of the identified bacteria with the geographical origin. Our study provides new insights into possible risk factors in dogs and evidenced discordant results between different tests, suggesting that a combination of serological and molecular assays is preferable for a correct diagnosis.

Human Tick-Borne Diseases and Advances in Anti-Tick Vaccine Approaches: A Comprehensive Review.

This comprehensive review explores the field of anti-tick vaccines, addressing their significance in combating tick-borne diseases of public health concern. The main objectives are to provide a brief epidemiology of diseases affecting humans and a thorough understanding of tick biology, traditional tick control methods, the development and mechanisms of anti-tick vaccines, their efficacy in field applications, associated challenges, and future prospects. Tick-borne diseases (TBDs) pose a significant and escalating threat to global health and the livestock industries due to the widespread distribution of ticks and the multitude of pathogens they transmit. Traditional tick control methods, such as acaricides and repellents, have limitations, including environmental concerns and the emergence of tick resistance. Anti-tick vaccines offer a promising alternative by targeting specific tick proteins crucial for feeding and pathogen transmission. Developing vaccines with antigens based on these essential proteins is likely to disrupt these processes. Indeed, anti-tick vaccines have shown efficacy in laboratory and field trials successfully implemented in livestock, reducing the prevalence of TBDs. However, some challenges still remain, including vaccine efficacy on different hosts, polymorphisms in ticks of the same species, and the economic considerations of adopting large-scale vaccine strategies. Emerging technologies and approaches hold promise for improving anti-tick vaccine development and expanding their impact on public health and agriculture.

Serological investigation of seven zoonotic pathogens in companion dogs in South Korea, 2018-2021.

Based on the current situation of Korean culture and society, the population of companion animals in South Korea is growing rapidly along with zoonotic risks. The current data regarding zoonotic infections in companion dogs reported in Korea is sparse. This study aims to investigate the seroprevalence of seven potential zoonotic pathogens in companion dogs in South Korea: Anaplasma phagocytophilum, Borrelia burgdoferi, Ehrlichia canis, Coxiella burnetii, Brucella canis, Leptospira spp. and canine influenza A virus. A total of 284 serum samples were collected from 2018 to 2021, and the immunoglobulin G (IgG) antibodies against 7 zoonotic pathogens were detected using enzyme-linked immunosorbent assays. Samples were divided into five groups and analysed based on age. IgG antibodies against six of the seven pathogens were detected. The highest seropositivity rate was detected for canine influenza A virus exposure (59.1%) for which the rates were the highest in dogs under 1 year old and declined with age. Positivity rates of the other pathogens were relatively low: 1.76% for Leptospira spp., 1.40% for A. phagocytophilum and E. canis, 1.06% for B. canis and 0.35% for B. burgdoferi. No antibodies against C. burnetii were detected in this study. The exposure of dogs in South Korea to six zoonotic pathogens was serologically confirmed, highlighting a potential risk for human infection. The zoonotic risk of companion dogs cannot be neglected, and implementation of One Health approach should be advocated to establish effective preventive measures.

Exploring the influence of host community composition on the outbreak potential of Anaplasma phagocytophilum and Borrelia burgdorferi s.l.

In large parts of the northern hemisphere, multiple deer species coexist, and management actions can strongly influence wild deer communities. Such changes may also indirectly influence other species in the community, such as small mammals and birds, because deer can have strong effects on their habitats and resources. Deer, small mammals and birds play an important role in the dynamics of tick-borne zoonotic diseases. It is, however, relatively underexplored how the abundance and composition of vertebrate communities may affect the outbreak potential, maintenance and circulation of tick-borne pathogens. In this study we focus on the outbreak potential by exploring how the basic reproduction number R0 for different tick-borne pathogens depends on host community composition. We used published data on co-varying roe deer (Capreolus capreolus) and fallow deer (Dama dama) densities following a hunting ban, and different small mammal and bird densities, to investigate how the change in host community influences the R0 of four tick-borne pathogens: one non-zoonotic, namely Anaplasma phagocytophilum ecotype 2, and three zoonotic, namely A. phagocytophilum ecotype 1, Borrelia afzelii and Borrelia garinii. We calculated R0 using a next generation matrix approach, and used elasticities to quantify the contributions to R0 of the different groups of host species. The value of R0 for A. phagocytophilum ecotype 1 was higher with high fallow deer density and low roe deer density, while it was the other way round for A. phagocytophilum ecotype 2. For B. afzelii, R0 was mostly related to the density of small mammals and for B. garinii it was mostly determined by bird density. Our results show that the effect of species composition is substantial in the outbreak potential of tick-borne pathogens. This implies that also management actions that change this composition, can (indirectly and unintentionally) affect the outbreak potential of tick-borne diseases.

Epidemiological and phylogenetic characteristics of emerging Anaplasma capra: A systematic review with modeling analysis.

Anaplasma capra, an emerging tick-borne pathogen, has caused a lot of concern since initially recognized in goats and patients in China in 2015, and has been reported in a wide range of domestic and wild animals as well as ticks worldwide, posing a threat to public health. In this systematic review, we established a comprehensive database to acquire the distribution and prevalence status of this pathogen, and collected all sequences of A. capra to summarize the details of genetic diversity by phylogenetic analysis. We also predicted the possible global distribution of A. capra by using ecological niche model. A. capra has been known to distribute in 18 countries across Asia, Europe and Africa. A total of 19 species of mammals from seven families have been reported as hosts, and domestic ruminants including goats, sheep and cattle were the major hosts. At least 8 tick species of 4 genera have been reported to carry A. capra, and Haemaphysalis longicornis was most commonly infected. Sheep and Rhipicephalus microplus had the highest positive rates among animals and ticks. Phylogenetic analysis based on gltA and groEL genes revealed that A. capra could primarily be divided into two clusters related to geographic location and animal hosts. The predictive model showed that the most suitable habitats for presence of A. capra were mainly located in Asia and eastern Europe. These cumulative data regarding A. capra of our study lay a foundation for the subsequent exploration of this emerging tick-borne pathogen.

Seroprevalence of Crimean Congo Hemorrhagic Fever Virus in Occupational Settings: Systematic Review and Meta-Analysis.

Crimean Congo Hemorrhagic Fever (CCHF) Virus can cause a serious human disease, with the case fatality ratio previously estimated to be 30-40%. Our study summarized seroprevalence data from occupational settings, focusing on the following occupational groups: animal handlers, abattoir workers, farmers, healthcare workers, veterinarians, rangers, and hunters. Systematic research was performed on three databases (PubMed, EMBASE, MedRxiv), and all studies reporting seroprevalence rates (IgG-positive status) for CCHF virus were retrieved and their results were reported, summarized, and compared. We identified a total of 33 articles, including a total of 20,195 samples, i.e., 13,197 workers from index occupational groups and 6998 individuals from the general population. Pooled seroprevalence rates ranged from 4.751% (95% confidence intervals (95% CI) 1.834 to 11.702) among animal handlers, to 3.403% (95% CI 2.44 to 3.932) for farmers, 2.737% (95% CI 0.896 to 8.054) among rangers and hunters, 1.900% (95% CI 0.738 to 4.808) for abattoir workers, and 0.644% (95% CI 0.223-1.849) for healthcare workers, with the lowest estimate found in veterinarians (0.283%, 95% CI 0.040-1.977). Seroprevalence rates for abattoir workers (odds ratio (OR) 4.198, 95% CI 1.060-16.464), animal handlers (OR 2.399, 95% CI 1.318-4.369), and farmers (OR 2.280, 95% CI 1.419 to 3.662) largely exceeded the official notification rates for CCHF in the general population. CCHF is reasonably underreported, and pooled estimates stress the importance of improving the adherence to personal protective equipment use and appropriate preventive habits.

Tick Densities and Infection Prevalence on Coastal Islands in Massachusetts, USA: Establishing a Baseline.

Tick-borne diseases and a tick-induced red meat allergy have become increasingly common in the northeastern USA and elsewhere. At the scale of local communities, few studies have documented tick densities or infection levels to characterize current conditions and provide a baseline for further monitoring. Using the town of Nantucket, MA, as a case study, we recorded tick densities by drag sampling along hiking trails in nature preserves on two islands. Nymphal blacklegged ticks (Ixodes scapularis Say) were most abundant at shadier sites and least common in grasslands and scrub oak thickets (Quercus ilicifolia). Lone star ticks (Amblyomma americanum L.) were common on Tuckernuck Island and rare on Nantucket Island, while both tick species were more numerous in 2021 compared to 2020 and 2022. We tested for pathogens in blacklegged nymphs at five sites over two years. In 2020 and 2021, infection levels among the four Nantucket Island sites averaged 10% vs. 19% for Borrelia burgdorferi, 11% vs. 15% for Babesia microti, and 17% (both years) for Anaplasma phagocytophilum, while corresponding levels were significantly greater on Tuckernuck in 2021. Our site-specific, quantitative approach represents a practical example of how potential exposure to tick-borne diseases can be monitored on a local scale.

Strict Conservation yet Non-Essential Nature of Plasmid Gene bba40 in the Lyme Disease Spirochete Borrelia burgdorferi.

The highly segmented genome of Borrelia burgdorferi, the tick-borne bacterium that causes Lyme disease, is composed of a linear chromosome and more than 20 co-existing endogenous plasmids. Many plasmid-borne genes are unique to B. burgdorferi and some have been shown to provide essential functions at discrete points of the infectious cycle between a tick vector and rodent host. In this study, we investigated the role of bba40, a highly conserved and differentially expressed gene on a ubiquitous linear plasmid of B. burgdorferi. In a prior genome-wide analysis, inactivation of bba40 by transposon insertion was linked with a noninfectious phenotype in mice, suggesting that conservation of the gene in the Lyme disease spirochete reflected a critical function of the encoded protein. To address this hypothesis, we moved the bba40::Tn allele into a similar wild-type background and compared the phenotypes of isogenic wild-type, mutant and complemented strains in vitro and throughout the in vivo mouse/tick infectious cycle. In contrast to the previous study, we identified no defect in the ability of the bba40 mutant to colonize the tick vector or murine host, or to be efficiently transmitted between them. We conclude that bba40 joins a growing list of unique, highly conserved, yet fully dispensable plasmid-borne genes of the Lyme disease spirochete. We infer that the experimental infectious cycle, while including the tick vector and murine host, lacks key selective forces imposed during the natural enzootic cycle. IMPORTANCE The key finding of this study contradicts our premise that the ubiquitous presence and strict sequence conservation of a unique gene in the Lyme disease spirochete, Borrelia burgdorferi, reflect a critical role in either the murine host or tick vector in which these bacteria are maintained in nature. Instead, the outcome of this investigation illustrates the inadequate nature of the experimental infectious cycle currently employed in the laboratory to fully model the enzootic cycle of the Lyme disease spirochete. This study also highlights the importance of complementation for accurate interpretation of mutant phenotypes in genetic studies of Borrelia burgdorferi.

Presence of Anaplasma phagocytophilum Ecotype I in UK Ruminants and Associated Zoonotic Risk.

Anaplasma phagocytophilum is the causative agent of tick-borne fever in sheep, pasture fever in cattle, and granulocytic anaplasmosis in humans. The increasing prevalence and transboundary spread of A. phagocytophilum in livestock, ticks, and wildlife in the UK poses a potential zoonotic risk that has yet to be estimated. Several ecotypes of A. phagocytophilum show variable zoonotic potential. To evaluate the possible risk associated with the transmission of A. phagocytophilum from ruminants to humans, the ecotype was determined by sequencing the groEL gene from 71 positive blood and tissue samples from UK ruminants. Thirty-four groEL sequences were obtained, fourteen of which were identified in multiple samples. Of the 13 nucleotide polymorphisms identified through pairwise comparison, all corresponded to synonymous substitutions. The subsequent phylogenetic estimation of the relationship with other European/world isolates indicated that all the groEL sequences clustered with other ecotype I sequences. The presence of ecotype I closely reflects that observed in ruminants in continental Europe and suggests a lower risk of zoonotic transmission from this reservoir.

Neoehrlichia mikurensis-A New Emerging Tick-Borne Pathogen in North-Eastern Poland?

Neoehrlichia mikurensis is a new emerging tick-borne Gram-negative bacterium, belonging to the family Anaplasmataceae, the main vector of which in Europe is the tick Ixodes ricinus. N. mikurensis is responsible for neoehrlichiosis, occurring mostly in patients with underlying diseases. In the present study, a total of 348 I. ricinus and Dermacentor reticulatus ticks collected in north-eastern Poland were analyzed for the prevalence of N. mikurensis. A total of 140 questing ticks (124 of I. ricinus ticks and 16 D. reticulatus) collected with the flagging method and 208 ticks (105 and 103 I. ricinus and D. reticulatus, respectively) removed from dogs were selected for the study. cDNA (questing ticks) and total DNA (questing and feeding ticks) were analyzed by qPCR targeting the 16S rRNA gene of N. mikurensis. Positive samples were further analyzed by nested PCR and sequencing. The prevalence differed between ticks collected from vegetation (19.3%; 27/140) and ticks removed from dogs (6.7%; 14/208). The presence of the pathogen in questing and feeding D. reticulatus ticks was proven in Poland for the first time. In summary, our research showed that infections of ticks of both the most common tick species I. ricinus and D. reticulatus in north-eastern Poland are present and ticks collected from urban areas were more often infected than ticks from suburban and natural areas. The detection of N. mikurensis in I. ricinus and D. reticulatus ticks from north-eastern Poland indicates potential transmission risk for tick-bitten humans at this latitude.

Voles, shrews and red squirrels as sources of tick blood meals and tick-borne pathogens on an island in southwestern Finland.

Molecular identification of the previous blood meal source of a questing tick (Acari: Ixodidae) from blood meal fragments was proposed a few decades ago. Following this, several blood meal assays have been developed and published, but none of them have been taken into widespread use. Recently, novel retrotransposon-based qPCR assays designed for detecting blood meal fragments of North American host species were published. We wanted to assess their function with host species present in Finland. Questing ticks were collected by cloth dragging in August-September 2021 from an island in southwestern Finland. DNA was extracted from Ixodes ricinus nymphs (n=438) and qPCR assays applied to identify larval blood meal sources (voles, shrews and red squirrels) and screen for several tick-borne human pathogens and other microbes with pathogenic potential [Borrelia spp. (including specific assays for Borrelia afzelii, Borrelia garinii, Borrelia valaisiana), Anaplasma phagocytophilum, Babesia spp., Rickettsia spp., and Neoehrlichia mikurensis]. The probability of a nymph having fed as larva on either a vole, shrew or red squirrel was 0.34 (0.30 - 0.38; 95% confidence interval). Bacteria of the genus Borrelia were the most common pathogens detected, with host-specific probabilities of carrying Borrelia of 0.30 (0.18 - 0.44) for nymphs that had fed on voles, 0.23 (0.14 - 0.35) for nymphs that had fed on shrews, and 0.42 (0.28 - 0.58) for nymphs that had fed on red squirrels. Other microbes were rarely acquired from these hosts, apart from N. mikurensis from voles. This study highlights that shrews and red squirrels may equal voles as blood meal sources for I. ricinus larvae. Overall, variation in proportions of blood meals provided by these animals may be high across even proximate study areas. All studied host species appeared to be important sources for particularly Borrelia afzelii, and voles also for N. mikurensis.

Detection of Tick-Borne Pathogens in Ticks from Cattle in Western Highlands of Cameroon.

This study aimed to detect and identify microorganisms in ticks collected in the Western Highlands of Cameroon. Quantitative real-time and standard PCR assays, coupled with sequencing, were used. A total of 944 ticks collected from cattle in five distinct sites in Cameroon were selected for the analyses. They belonged to five genera (Amblyomma, Hyalomma, Rhipicephalus, Haemaphysalis, and Ixodes) and twelve species. Real-time PCR revealed that 23% (n = 218) of the ticks were positive for Rickettsia spp., 15% (n = 141) for bacteria of the Anaplasmataceae family, 3% (n = 29) for Piroplasmida, 0.5% (n = 5) for Coxiella burnetii, 0.4% (n = 4) for Borrelia spp., and 0.2% (n = 2) for Bartonella spp. The co-infection rate (3.4%, n = 32) involved mainly Rickettsia spp. and Anaplasmataceae. Of the Rickettsia spp. positive ticks, the targeted PCR and sequencing yielded Rickettsia africae (78.9%), Rickettsia aeschlimannii (6.4%), Rickettsia massiliae (7.8%), Candidatus Rickettsia barbariae (0.9%), and Rickettsia sp. (0.9%). Anaplasmataceae included Anaplasma marginale (4.3%), Anaplasma platys (1.4%), Anaplasma centrale (0.7%), Ehrlichia ruminantium (0.7%), Wolbachia sp., Candidatus Ehrlichia rustica (13.5%), Candidatus Ehrlichia urmitei (7%), and an uncultured Ehrlichia sp. (4.2%). Borrelia theileri was identified in one Rhipicephalus microplus tick. Unfortunately, Piroplasmida could not be identified to the species level. This study demonstrates that in Cameroon, ticks harbour a wide variety of microorganisms and present a risk of zoonotic diseases.

Environmental factors driving fine-scale ixodid tick abundance patterns.

Tick abundance is an essential demographic parameter to infer tick-borne pathogen transmission risks. Spatiotemporal patterns of tick abundance are heterogeneous, so its determinants at small spatial scales need to be understood to reduce their negative effects on hosts. Current knowledge of these determinants is scarce, especially in Mediterranean environments, limiting the possibilities for designing efficient tick control strategies. With the goal of unravelling tick abundance determinants and informing new tick management strategies, we estimated tick burdens on 1965 wild ungulates in Doñana National Park, Spain, annually between 2010 and 2020. Under the hypothesis of a predominant host influence on tick abundance, we modelled the burdens of Rhipicephalus annulatus, Hyalomma lusitanicum, and Ixodes ricinus with relevant predictors grouped into four factors: i) environment; ii) host population; iii) host individual; and iv) land-use. Generalized linear mixed models with a zero-inflated negative binomial distribution were built. Additionally, we analysed the differential contribution to abundance of each factor by deviance partitioning. We finally estimated the similarity in the environmental space of tick species by analysing their niche overlap with the environmental principal component analysis method. Our work hypothesis was confirmed for R. annulatus and H. lusitanicum, but we found that tick abundance at a fine spatial scale is jointly driven by multiple drivers, including all four factors considered in this study. This result points out that understanding the demography of ticks is a complex multifactorial issue, even at small spatial scales. We found no niche differences between the three tick species at the study spatial scale, thus showing similar host and environmental dependencies. Overall results identify that host aggregation areas displaying environmentally favourable traits for ticks are relevant tick and vector-borne pathogen transmission hotspots. Our findings will facilitate the design of new strategies to reduce the negative effects of tick parasitism.

Diversity of Tick-Borne Pathogens in Tick Larvae Feeding on Breeding Birds in France.

Birds play a role in maintaining tick-borne diseases by contributing to the multiplication of ticks and pathogens on a local scale during the breeding season. In the present study, we describe the diversity of tick and pathogen species of medical and veterinary importance in Europe hosted by 1040 captured birds (56 species) during their breeding season in France. Of the 3114 ticks collected, Ixodes ricinus was the most prevalent species (89.5%), followed by I. frontalis (0.8%), I. arboricola (0.7%), Haemaphysalis concinna (0.5%), H. punctata (0.5%), Hyalomma spp. (0.2%), and Rhipicephalus spp. (0.06%). Because they may be representative of the bird infection status for some pathogen species, 1106 engorged tick larvae were screened for pathogens. Borrelia burgdorferi sensu lato was the most prevalent pathogen genus in bird-feeding larvae (11.7%), followed by Rickettsia spp. (7.4%), Anaplasma spp. (5.7%), Babesia spp. (2.3%), Ehrlichia spp. (1.4%), and B. miyamotoi (1%). Turdidae birds (Turdus merula and T. philomelos), Troglodytes troglodytes, and Anthus trivialis had a significantly higher prevalence of B. burgdorferi s.l.-infected larvae than other pathogen genera. This suggests that these bird species could act as reservoir hosts for B. burgdorferi s.l. during their breeding season, and thus play an important role in acarological risk.

Predicting habitat suitability for Ixodes ricinus and Ixodes persulcatus ticks in Finland.

BACKGROUND: Ticks are responsible for transmitting several notable pathogens worldwide. Finland lies in a zone where two human-biting tick species co-occur: Ixodes ricinus and Ixodes persulcatus. Tick densities have increased in boreal regions worldwide during past decades, and tick-borne pathogens have been identified as one of the major threats to public health in the face of climate change. METHODS: We used species distribution modelling techniques to predict the distributions of I. ricinus and I. persulcatus, using aggregated historical data from 2014 to 2020 and new tick occurrence data from 2021. By aiming to fill the gaps in tick occurrence data, we created a new sampling strategy across Finland. We also screened for tick-borne encephalitis virus (TBEV) and Borrelia from the newly collected ticks. Climate, land use and vegetation data, and population densities of the tick hosts were used in various combinations on four data sets to estimate tick species' distributions across mainland Finland with a 1-km resolution. RESULTS: In the 2021 survey, 89 new locations were sampled of which 25 new presences and 63 absences were found for I. ricinus and one new presence and 88 absences for I. persulcatus. A total of 502 ticks were collected and analysed; no ticks were positive for TBEV, while 56 (47%) of the 120 pools, including adult, nymph, and larva pools, were positive for Borrelia (minimum infection rate 11.2%, respectively). Our prediction results demonstrate that two combined predictor data sets based on ensemble mean models yielded the highest predictive accuracy for both I. ricinus (AUC = 0.91, 0.94) and I. persulcatus (AUC = 0.93, 0.96). The suitable habitats for I. ricinus were determined by higher relative humidity, air temperature, precipitation sum, and middle-infrared reflectance levels and higher densities of white-tailed deer, European hare, and red fox. For I. persulcatus, locations with greater precipitation and air temperature and higher white-tailed deer, roe deer, and mountain hare densities were associated with higher occurrence probabilities. Suitable habitats for I. ricinus ranged from southern Finland up to Central Ostrobothnia and North Karelia, excluding areas in Ostrobothnia and Pirkanmaa. For I. persulcatus, suitable areas were located along the western coast from Ostrobothnia to southern Lapland, in North Karelia, North Savo, Kainuu, and areas in Pirkanmaa and Päijät-Häme. CONCLUSIONS: This is the first study conducted in Finland that estimates potential tick species distributions using environmental and host data. Our results can be utilized in vector control strategies, as supporting material in recommendations issued by public health authorities, and as predictor data for modelling the risk for tick-borne diseases.