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Background: Ticks carry a variety of microorganisms, some of which are pathogenic to humans. The human risk of tick-borne diseases depends on, among others, the prevalence of pathogens in ticks biting humans. To follow-up on this prevalence over time, a Belgian study from 2017 was repeated in 2021. Methods: During the tick season 2021, citizens were invited to have ticks removed from their skin, send them and fill in a short questionnaire on an existing citizen science platform for the notification of tick bites (TekenNet). Ticks were morphologically identified to species and life stage level and screened using multiplex qPCR targeting, among others, Borrelia burgdorferi (sensu lato), Anaplasma phagocytophilum, Borrelia miyamotoi, Neoehrlichia mikurensis, Babesia spp., Rickettsia helvetica and tick-borne encephalitis virus (TBEV). The same methodology as in 2017 was used. Results: In 2021, the same tick species as in 2017 were identified in similar proportions; of 1094 ticks, 98.7% were Ixodes ricinus, 0.8% Ixodes hexagonus and 0.5% Dermacentor reticulatus. A total of 928 nymphs and adults could be screened for the presence of pathogens. Borrelia burgdorferi (s.l.) was detected in 9.9% (95% CI 8.2–12.0%), which is significantly lower than the prevalence of 13.9% (95% CI 12.2–15.7%) in 2017 (P = 0.004). The prevalences of A. phagocytophilum (4.7%; 95% CI 3.5–6.3%) and R. helvetica (13.3%; 95% CI 11.2–15.6%) in 2021 were significantly higher compared to 2017 (1.8%; 95% CI 1.3–2.7% and 6.8%; 95% CI 5.6–8.2% respectively) (P < 0.001 for both). For the other pathogens tested, no statistical differences compared to 2017 were found, with prevalences ranging between 1.5 and 2.9% in 2021. Rickettsia raoultii was again found in D. reticulatus ticks (n = 3/5 in 2021). Similar to 2017, no TBEV was detected in the ticks. Co-infections were found in 5.1% of ticks. When combining co-infection occurrence in 2017 and 2021, a positive correlation was observed between B. burgdorferi (s.l.) and N. mikurensis and B. burgdorferi (s.l.) and B. miyamotoi (P < 0.001 for both). Conclusions: Although the 2021 prevalences fell within expectations, differences were found compared to 2017. Further research to understand the explanations behind these differences is needed. [ABSTRACT FROM AUTHOR]

Severe babesiosis with 9.8% parasitemia was diagnosed in a patient in the Netherlands who had previously undergone splenectomy. We confirmed Babesia venatorum using PCR and sequencing. B. venatorum was also the most prevalent species in Ixodes ricinus ticks collected around the patient’s home. Our findings warrant awareness for severe babesiosis in similar patients. [ABSTRACT FROM AUTHOR]

Background: Bartonella spp. infect a variety of vertebrates throughout the world, with generally high prevalence. Several Bartonella spp. are known to cause diverse clinical manifestations in humans and have been recognized as emerging pathogens. These bacteria are mainly transmitted by blood-sucking arthropods, such as fleas and lice. The role of ticks in the transmission of Bartonella spp. is unclear. Methods: A recently developed quadruplex polymerase chain reaction (PCR) amplicon next-generation sequencing approach that targets Bartonella-specific fragments on gltA, ssrA, rpoB, and groEL was applied to test host-seeking Ixodes scapularis ticks (n = 1641; consisting of 886 nymphs and 755 adults) collected in 23 states of the eastern half of the United States and Ixodes pacificus ticks (n = 966; all nymphs) collected in California in the western United States for the presence of Bartonella DNA. These species were selected because they are common human biters and serve as vectors of pathogens causing the greatest number of vector-borne diseases in the United States. Results: No Bartonella DNA was detected in any of the ticks tested by any target. Conclusions: Owing to the lack of Bartonella detection in a large number of host-seeking Ixodes spp. ticks tested across a broad geographical region, our results strongly suggest that I. scapularis and I. pacificus are unlikely to contribute more than minimally, if at all, to the transmission of Bartonella spp. [ABSTRACT FROM AUTHOR]

Tick‐borne encephalitis (TBE) virus is considered the medically most important arthropod‐borne virus in Europe. Although TBE is endemic throughout central Europe, ticks and rodents determine its maintenance in small, difficult‐to‐assess, natural foci. We investigated the interrelation between the population genetics of the main TBE virus (TBEV) vector tick (Ixodes ricinus), the most important reservoir host (Myodes glareolus, syn. Clethrionomys glareolus), and TBEV. Rodents and ticks were sampled on 15 sites within an exploratory study area, which has been screened regularly for TBEV occurrence in ticks for more than 10 years. On all 15 sites, ticks and bank voles were sampled, screened for TBEV presence via serology and RT‐PCR, and genetically examined. Moreover, TBEV isolates derived from these analyses were sequenced. In long‐term TBEV foci bank vole populations show extraordinary genetic constitutions, leading to a particular population structure, whereas ticks revealed a panmictic genetic structure overall sampling sites. Landscape genetics and habitat connectivity modeling (analysis of isolation by resistance) showed no landscape‐related barriers explaining the genetic structure of the bank vole populations. The results suggest that bank voles do not simply serve as TBEV reservoirs, but their genetic composition appears to have a significant influence on establishing and maintaining long‐term natural TBEV foci, whereas the genetic structure of TBEV's main vector I. ricinus does not play an important role in the sustainability of long‐term TBEV foci. A thorough investigation of how and to which extent TBEV and M. glareolus genetics are associated is needed to further unravel the underlying mechanisms. [ABSTRACT FROM AUTHOR]

Simple Summary: Dermacentor reticulatus is an arthropod vector with great medical and veterinary importance. Its wide distribution and biological characteristics determine its important role in the circulation of pathogens in the parasite–host system. Its occurrence range is divided into western and eastern populations, which are separated from each other by the so-called Dermacentor-free zone localized in central Poland. This study aimed to estimate the potential epidemiological significance of D. reticulatus in the new endemic focus west of the Vistula River (Upper Silesia, central–southern Poland) and its co-occurrence with Ixodes ricinus. The molecular studies revealed the presence of Rickettsia spp. in 23.8% of the D. reticulatus specimens. In turn, 94.1% of the I. ricinus adults were infected with B. burgdorferi s.l., 11.7% with Babesia spp., and 5.8% with Rickettsia spp. Polymicrobial infections were noted in 17.6% of the I. ricinus. Our finding emphasizes the risk of infestation by both tick species and the risk of tick-borne infections in an area previously thought to be free of Dermacentor ticks. It is necessary to enhance medical and veterinary services for the more efficient diagnosis and prevention of tick-borne diseases in this region. Dermacentor reticulatus (Acari: Ixodidae) is an important arthropod vector in medical and veterinary contexts. Its geographic range is divided into western and eastern populations separated by a "Dermacentor-free zone" in central Poland. Recent faunistic studies showed a new endemic locality of the species in Upper Silesia to the west of the Vistula River (central–southern Poland) and its co-occurrence with I. ricinus. The prevalence of five tick-borne pathogens (TBPs), e.g., B. burgdorferi s.l., Bartonella spp., Rickettsia spp., and Babesia spp., in the ticks was assessed with polymerase chain reaction (PCR) methods. The molecular studies revealed the presence of Rickettsia spp. in 23.8% of the D. reticulatus specimens. In turn, 94.1% of the I. ricinus adults were infected with B. burgdorferi s.l., 11.7 % with Babesia spp., and 5.8% with Rickettsia spp. Coinfections with two TBPs were noted in 17.6% of the I. ricinus. These findings highlight not only the risk of infestation by both tick species in an area previously considered Dermacentor-free, but also the high prevalence of TBPs in the study area. Increased focus on medical and veterinary services appears necessary to diagnose and prevent tick-borne diseases in this region. [ABSTRACT FROM AUTHOR]

Simple Summary: We conducted a tick surveillance study in northwestern Spain with the aim of identifying tick-associated Rickettsia species among ticks removed from humans and to track how tick populations and their associated pathogens have changed over the years. Five genera and eight species of ticks were collected, with Ixodes ricinus being the most frequently found. By comparing our data with previous reports, a clear geographic and seasonal expansion of tick populations and the associated Rickettsia species is observed, indicating that the spatiotemporal patterns of ticks and bacteria have changed over the years. Considering the medical relevance of tick-borne rickettsioses, evaluating the infection risk to humans in tick-infested areas, as well as detecting spreading species, is essential for establishing adequate prevention and control measures. Tick-borne rickettsioses (TBRs) are distributed worldwide and are recognized as important emerging vector-borne zoonotic diseases in Europe. The aim of this study was to identify tick-associated Rickettsia among ticks removed from humans, and to track how tick populations and their associated pathogens have changed over the years. For this purpose, we conducted a tick surveillance study in northwestern Spain between 2018 and 2022. Ticks were morphologically identified and analyzed for the presence of rickettsial pathogens through the amplification of the citrate synthase (gltA) and the outer membrane protein A (ompA) genes. PCR products were sequenced and subjected to phylogenetic analyses. We collected 7397 ticks, with Ixodes ricinus being the species most frequently isolated. Based on the PCR results, Rickettsia DNA was detected in 1177 (15.91%) ticks, and 10 members of Rickettsia were identified: R. aeschlimannii, R. conorii subsp. conorii, R. conorii subsp. raoultii, R. massiliae, R. monacensis, R. sibirica subsp. mongolitimonae, R. slovaca, R. helvetica, Candidatus R. barbariae, and Candidatus R. rioja. Some of these Rickettsia have gone previously undetected in the study region. There is clear geographic and seasonal expansion not only of tick populations, but also of the associated Rickettsia. The comparison of our data with those obtained years ago provides a clear idea of how the spatiotemporal distributions of ticks and their associated Rickettsiae have changed over the years. [ABSTRACT FROM AUTHOR]

Simple Summary: Ticks are spreading at high altitudes in mountain areas and can come into contact with previously unexposed or poorly exposed hosts, such as Alpine ibex (Capra ibex). We collected ticks on ibex and vegetation in the northwestern Italian Alps and tested them for tick-borne pathogens. Ixodes ricinus was the most abundant tick species collected and was infected with Borrelia burgdorferi s.l., Rickettsia spp. and Anaplasma phagocytophilum. Our results suggest that Alpine ibex, like other wild ungulate species, may act as a reservoir for A. phagocytophilum while being an incompetent reservoir for the Lyme borreliosis agent. Future studies should monitor the possible impacts of ticks and transmitted pathogens on the health and conservation of ibex. In the Maritime Alps (northwestern Italy), we collected ticks from vegetation and Alpine ibex (Capra ibex). Ixodes ricinus was the most abundant species in the study area, questing up to 1824 m a.s.l. and infesting 28 out of 72 ibexes. Haemaphysalis punctata, H. sulcata and Dermacentor marginatus were also collected. The abundance of questing ticks significantly decreased with altitude, with beechwoods being the preferred habitat. By PCR, we identified Borrelia burgdorferi s.l. in questing I. ricinus (28.3%; 95%CI: 19.4–38.6) but not in specimens collected from animals. Rickettsia spp. infected both questing (20.6%; 95%CI: 12.9–30.3) and on-host (30.2%; 95%CI: 21.2–40.4) I. ricinus. Anaplasma phagocytophilum was detected in 4.3% (95%CI: 1.2–10.8) of questing I. ricinus and in 45.3% (95%CI: 34.6–56.4) of I. ricinus collected from ibex. Female I. ricinus collected on animals were significantly more infected with A. phagocytophilum than females collected from vegetation (OR = 11.7; 95%CI: 3.8–48.1). By amplifying and sequencing a fragment of the groEL gene, we identified 13 groEL haplotypes, clustering with ecotypes I and II; ecotype I, prevalent in our sample, is considered zoonotic. Our study demonstrates the presence of different tick-borne zoonotic agents in the study area, encompassing a wide altitudinal range, as confirmed by the ticks found on ibex, a typical mountain-dwelling mammal. The results also confirm the altitudinal range expansion of ticks and associated pathogens in the Alps and suggest that Alpine ibex may act as a reservoir for A. phagocytophilum, as do other wild ungulate species. [ABSTRACT FROM AUTHOR]

Simple Summary: Ticks are ectoparasites with medical significance. They inhabit diverse environments and maintain close interactions with numerous vertebrate hosts. Ixodes ticks can transmit various pathogens to animals and humans. The aim here was to examine Ixodes ticks from Bosnia and Herzegovina to check for specific pathogens. This study found Rickettsia spp., Babesia spp., Anaplasma phagocytophilum, and Borrelia burgdorferi sensu lato in ticks from domestic animals. These findings highlight the need for the ongoing monitoring of ticks and tick-borne pathogens to protect animal and public health. Additionally, this study provides valuable insights into the occurrence and spread of these pathogens, emphasizing the importance of broader surveillance and control measures. Effective prevention, surveillance, and control of tick-borne diseases require urgent regional and international collaboration. Limited information is available regarding the presence of tick-borne pathogens and their distribution within Ixodes species in Bosnia and Herzegovina. This study aimed to identify Rickettsia spp., Babesia spp., Anaplasma phagocytophilum, and Borrelia burgdorferi sensu lato (s.l.) in Ixodes ticks collected from domestic and wild animals and vegetation in different regions across Bosnia and Herzegovina. A total of 7438 adult ticks, including 4526 Ixodes ricinus, Ixodes canisuga, and Ixodes hexagonus, were collected. Real-time PCR screening of 450 pooled I. ricinus samples revealed a 22.1% infection rate with at least one pathogen. Rickettsia spp. (6.3%) were found in ticks from dogs, cats, and goats, Babesia spp. (3.1%) in ticks from dogs and cattle, A. phagocytophilum (8.8%) in ticks from dogs, goats, and cattle, and B. burgdorferi s.l. (3.4%) in ticks from dogs and cats. Mixed infections with B. burgdorferi s.l. and A. phagocytophilum, as well as B. burgdorferi s.l. and Rickettsia spp., were found in two pools of I. ricinus from dogs and cats, respectively. Additionally, co-infection with Rickettsia spp. and A. phagocytophilum was confirmed in three tick pools from dogs and goats. Each tick from these pooled samples was individually retested to confirm the presence of pathogens. In the examined pooled samples of I. canisuga (1) and I. hexagonus (6), none of the tested pathogens were detected. Our findings represent the first detection of Rickettsia spp., Babesia spp., A. phagocytophilum, and B. burgdorferi s.l. in I. ricinus collected from domestic animals and vegetation in Bosnia and Herzegovina. Considering the established infection rates, the detection of tick-borne pathogens in adult ticks collected from domestic animals and vegetation enriches the current knowledge of the presence of tick-borne pathogens at the local, regional, national, and broader levels. [ABSTRACT FROM AUTHOR]

Usutu virus (USUV) is an emerging flavivirus that is maintained in an enzootic cycle with mosquitoes as vectors and birds as amplifying hosts. In Europe, the virus has caused mass mortality of wild birds, mainly among Common Blackbird (Turdus merula) populations. While mosquitoes are the primary vectors for USUV, Common Blackbirds and other avian species are exposed to other arthropod ectoparasites, such as ticks. It is unknown, however, if ticks can maintain and transmit USUV. We addressed this question using in vitro and in vivo experiments and field collected data. USUV replicated in IRE/CTVM19 Ixodes ricinus tick cells and in injected ticks. Moreover, I. ricinus nymphs acquired the virus via artificial membrane blood-feeding and maintained the virus for at least 70 days. Transstadial transmission of USUV from nymphs to adults was confirmed in 4.9% of the ticks. USUV disseminated from the midgut to the haemocoel, and was transmitted via the saliva of the tick during artificial membrane blood-feeding. We further explored the role of ticks by monitoring USUV in questing ticks and in ticks feeding on wild birds in the Netherlands between 2016 and 2019. In total, 622 wild birds and the Ixodes ticks they carried were tested for USUV RNA. Of these birds, 48 (7.7%) carried USUV-positive ticks. The presence of negative-sense USUV RNA in ticks, as confirmed via small RNA-sequencing, showed active virus replication. In contrast, we did not detect USUV in 15,381 questing ticks collected in 2017 and 2019. We conclude that I. ricinus can be infected with USUV and can transstadially and horizontally transmit USUV. However, in comparison to mosquito-borne transmission, the role of I. ricinus ticks in the epidemiology of USUV is expected to be minor. Author summary: Usutu virus (USUV) is a virus primarily circulating between birds and mosquitoes. Other blood-feeding arthropods, such as ticks, feed on birds. However, it is unknown whether ticks can be infected with USUV and transmit the virus to birds or other animals. Therefore, we infected a tick cell line and ticks collected from the field with USUV. We showed that the virus replicates in tick cells, as well as in field-collected ticks. Moreover, we showed that USUV disseminated through the tick body and was excreted with the saliva of ticks during artificial feeding. We further investigated if ticks are infected with USUV while feeding on wild birds and when searching for a host in the vegetation. We found that 7.7% of tick-infested birds carried USUV positive ticks, while USUV was not detected in questing ticks. Our findings provide evidence that ticks can be infected with USUV and might transmit the virus to another host, but the role of ticks in the transmission of USUV is expected to be minor. [ABSTRACT FROM AUTHOR]

Ticks are blood‐sucking parasite and transmit pathogens to humans, domestic and wild animals. Ticks are seriously damage the production of sheep and goats, resulting in heavy economic loss to farmers. The present study investigated microscopic and molecular identification of ticks and tick‐borne anaplasmosis infesting sheep and goats in three agro‐ecological zones of Pakistan. The collected ticks were identified at species level based on their morphological features through microscopy by using taxonomic keys. Four Rhipicephalus (R.) species namely: R. guilhoni Rh. geigyi, R. annulatus, R. pulchellus, two Haemaphysalis (Hae) spp. Hae. sulcate, Hae. punctate, one Ixodes ricinus and Hyalomma anatolicum were identified. The morphological variations in their body sizes, scutal pattern, female genital opening, spiracular plate length and male adanal plate were recorded. Furthermore, we assessed the effectiveness of utilizing the 16S rRNA gene sequence to differentiate Anaplasma samples at the species level. Microscopically suspected positive blood samples for Anaplasma were subjected to DNA extraction. Anaplasma was observed in the blood of 14 animals. The sequence similarity between Anaplasma marginale and the reference genome were 99.25%, while below this value indicating a distinct species. Phylogenetic analysis revealed close similarity with A. marginale isolates from Iran, Vietnam and Pakistan This study contributes to the epidemiology of ticks and TBPs in the KP province and highlights the importance of proactive TBP surveillance in livestock. [ABSTRACT FROM AUTHOR]

Aims: Urban green spaces are locations of maximal human activity, forming areas of enhanced risk for tick‐borne disease (TBD) transmission. Being also limited in spatial scale, green spaces form prime targets for control schemes aiming to reduce TBD risk. However, for effective control, the key species maintaining local tick and tick‐borne pathogen (TBP) populations must be identified. To determine how patterns of host utilization vary spatially, we utilized blood meal analysis to study the contributions of voles, shrews, squirrels, leporids and cervids towards blood meals and the acquisition of TBPs of juvenile Ixodes ricinus in urban and sylvatic areas in Finland. Methods and Results: A total of 1084 nymphs were collected from the capital city of Finland, Helsinki and from a sylvatic island in southwestern Finland, and subjected to qPCR analysis to identify DNA remnants of the previous host. We found significant differences in host contributions between urban and sylvatic environments. Specifically, squirrels and leporids were more common hosts in urban habitats, whereas cervids and voles were more common in sylvatic habitats. In addition to providing 18.4% of larval blood meals in urban habitats, red squirrels were identified as the source of 28.6% (n = 48) of Borrelia afzelii detections and 58.1% (n = 18) of Borrelia burgdorferi sensu stricto detections, indicating an important role for local enzootic cycles. Conclusions: Our study highlights that the key hosts maintaining tick and TBP populations may be different in urban and sylvatic habitats. Likewise, hosts generally perceived as important for upkeep may have limited importance in urban environments. Consequently, targeting control schemes based on off‐site data of host importance may lead to suboptimal results. [ABSTRACT FROM AUTHOR]

Identifying hybridization between common pathogen vectors is essential due to the major public health implications through risks associated with hybrid's enhanced pathogen transmission potential. The hard‐ticks Ixodes ricinus and Ixodes persulcatus are the two most common vectors of tick‐borne pathogens that affect human and animal health in Europe. Ixodes ricinus is a known native species in Finland with a well‐known distribution, whereas I. persulcatus has expanded in range and abundance over the past 60 years, and currently it appears the most common tick species in certain areas in Finland. Here we used double‐digest restriction site‐associated DNA (ddRAD) sequencing on 186 ticks (morphologically identified as 92 I. ricinus, and 94 I. persulcatus) collected across Finland to investigate whether RAD generated single nucleotide polymorphisms (SNPs) can discriminate tick species and identify potential hybridization events. Two different clustering methods were used to assign specific species based on how they clustered and identified hybrids among them. We were able to discriminate between the two tick species and identified 11 putative hybrids with admixed genomic proportions ranging from approximately 24 to 76 percent. Four of these hybrids were morphologically identified as I. ricinus while the remaining seven were identified as I. persulcatus. Our results thus indicate that RAD SNPs are robust in identifying both species of the ticks as well as putative hybrids. These results further suggest ongoing hybridization between I. ricinus and I. persulcatus in their natural populations in Finland. The unique ability of RAD markers to discriminate between tick species and hybrids adds a useful aspect to tick evolutionary studies. Our findings align with previous studies and suggest a shared evolutionary history between the species, with instances of individuals possessing a considerable proportion of the other species' genome. This study is a significant step in understanding the formation of hybridization zones due to range expansion potentially associated with climate change. [ABSTRACT FROM AUTHOR]

Simple Summary: This study shows the common occurrence of entomopathogenic fungi (EF) in the habitats of Ixodes ricinus, which indicates the necessity to consider EF as a potential factor influencing the tick population. However, the obtained results highlight the need for further research to fully understand the interaction between soil microorganisms and tick occurrence. (1) Background: In addition to the microclimate, host availability, and tick microbiota, soil environmental microorganisms can affect tick populations. This study aimed to (1) determine the presence and diversity of entomopathogenic fungi (EF) in forests, where ticks are abundant, and (2) estimate the effectiveness of the isolated EF strains against Ixodes ricinus. (2) Methods: EF were isolated using the trap insect method from soil collected from tick sites. A bioassay was used to estimate the effectiveness of EF against ticks. (3) Results: The presence of EF was found in all tested forest habitat types. A total of 53 strains belonging to the genera Metarhizium, Beauveria, and Isaria were isolated. All the six strains subjected to the bioassay showed potential efficacy against both adult and nymphal stages of I. ricinus; however, the strains differed in their effectiveness. The most effective isolate against I. ricinus was the soil environmental strain of Metarhizium anisopliae. (4) Conclusion: The study indicates that tick habitats can be the source of entomopathogenic fungi, which have a lethal effect on ticks, as demonstrated in preliminary laboratory tests with I. ricinus. However, for practical use, extensive field tests and further research on application methods and long-term effects are necessary to develop effective and sustainable tick management strategies. [ABSTRACT FROM AUTHOR]

Background: Ixodes inopinatus was described from Spain on the basis of morphology and partial sequencing of 16S ribosomal DNA. However, several studies suggested that morphological differences between I. inopinatus and Ixodes ricinus are minimal and that 16S rDNA lacks the power to distinguish the two species. Furthermore, nuclear and mitochondrial markers indicated evidence of hybridization between I. inopinatus and I. ricinus. In this study, we tested our hypothesis on tick dispersal from North Africa to Southern Europe and determined the prevalence of selected tick-borne pathogens (TBPs) in I. inopinatus, I. ricinus, and their hybrids. Methods: Ticks were collected in Italy and Algeria by flagging, identified by sequencing of partial TROSPA and COI genes, and screened for Borrelia burgdorferi s.l., B. miyamotoi, Rickettsia spp., and Anaplasma phagocytophilum by polymerase chain reaction and sequencing of specific markers. Results: Out of the 380 ticks, in Italy, 92 were I. ricinus, 3 were I. inopinatus, and 136 were hybrids of the two species. All 149 ticks from Algeria were I. inopinatus. Overall, 60% of ticks were positive for at least one TBP. Borrelia burgdorferi s.l. was detected in 19.5% of ticks, and it was significantly more prevalent in Ixodes ticks from Algeria than in ticks from Italy. Prevalence of Rickettsia spotted fever group (SFG) was 51.1%, with significantly greater prevalence in ticks from Algeria than in ticks from Italy. Borrelia miyamotoi and A. phagocytophilum were detected in low prevalence (0.9% and 5.2%, respectively) and only in ticks from Italy. Conclusions: This study indicates that I. inopinatus is a dominant species in Algeria, while I. ricinus and hybrids were common in Italy. The higher prevalence of B. burgdorferi s.l. and Rickettsia SFG in I. inopinatus compared with that in I. ricinus might be due to geographical and ecological differences between these two tick species. The role of I. inopinatus in the epidemiology of TBPs needs further investigation in the Mediterranean Basin. [ABSTRACT FROM AUTHOR]

Background: Many pathogens and parasites can infect multiple host species, and the competence of different hosts as pathogen reservoirs is key to understanding their epidemiology. Small mammals are important hosts for the instar stages of Ixodes ricinus ticks, the principal vector of Lyme disease in Europe. Small mammals also act as reservoirs of Borrelia afzelii, the most common genospecies of the Borrelia burgdorferi sensu lato (s.l.) spirochetes causing Lyme disease in Europe. However, we lack quantitative estimates on whether different small mammal species are equally suitable hosts for feeding I. ricinus and whether they show differences in pathogen transmission from host to tick. Methods: Here, we analysed the feeding success and prevalence of B. burgdorferi s.l. infections in 12,987 instar I. ricinus found on captured small mammals with known infection status in Norway (2018–2022). Results: We found that larvae were more likely to acquire a blood meal from common shrews (Sorex araneus, 46%) compared to bank voles (Myodes glareolus, 31%) and wood mice (Apodemus sylvaticus, 36%). Nymphs tended to be more likely to acquire a blood meal from wood mice (66%) compared to bank voles (54%). Common shrews harboured few nymphs (n=19). Furthermore, we found that larvae feeding on infected bank voles (11%) were more likely to be infected with B. burgdorferi s.l. than larvae on infected common shrews (7%) or wood mice (4%). Conclusions: Our study provides quantitative evidence of differences in suitability for the instar stages of I. ricinus across taxa of small mammals and highlights how even known small mammal host species can differ in their ability to feed ticks and infect larval ticks with the pathogen causing Lyme disease. [ABSTRACT FROM AUTHOR]

(1) Background: Tick-borne encephalitis (TBE) is the most important tick-borne viral disease in Eurasia, although effective vaccines are available. Caused by the tick-borne encephalitis virus (TBEV, syn. Orthoflavivirus encephalitidis), in Europe, it is transmitted by ticks like Ixodes ricinus and Dermacentor reticulatus. TBEV circulates in natural foci, making it endemic to specific regions, such as southern Germany and northeastern Poland. Our study aimed to identify new TBEV natural foci and genetically characterize strains in ticks in previously nonendemic areas in Eastern Germany and Western Poland. (2) Methods: Ticks were collected from vegetation in areas reported by TBE patients. After identification, ticks were tested for TBEV in pools of a maximum of 10 specimens using real-time RT-PCR. From the positive TBEV samples, E genes were sequenced. (3) Results: Among 8400 ticks from 19 sites, I. ricinus (n = 4784; 56.9%) was predominant, followed by D. reticulatus (n = 3506; 41.7%), Haemaphysalis concinna (n = 108; 1.3%), and I. frontalis (n = 2; <0.1%). TBEV was detected in 19 pools originating in six sites. The phylogenetic analyses revealed that TBEV strains from Germany and Poland clustered with other German strains, as well as those from Finland and Estonia. (4) Conclusions: Although there are still only a few cases are reported from these areas, people spending much time outdoors should consider TBE vaccination. [ABSTRACT FROM AUTHOR]

Cell cultures derived from ticks have become a commonly used tool for the isolation and study of tick-borne pathogens and tick biology. The IRE/CTVM19 cell line, originating from embryos of Ixodes ricinus, is one such line. Previously, reovirus-like particles, as well as sequences with similarity to rhabdoviruses and iflaviruses, were detected in the IRE/CTVM19 cell line, suggesting the presence of multiple persisting viruses. Subsequently, the full genome of an IRE/CTVM19-associated rhabdovirus was recovered from a cell culture during the isolation of the Alongshan virus. In the current work, we used high-throughput sequencing to describe a virome of the IRE/CTVM19 cell line. In addition to the previously detected IRE/CTVM19-associated rhabdovirus, two rhabdoviruses were detected: Chimay rhabdovirus and Norway mononegavirus 1. In the follow-up experiments, we were able to detect both positive and negative RNA strands of the IRE/CTVM19-associated rhabdovirus and Norway mononegavirus 1 in the IRE/CTVM19 cells, suggesting their active replication in the cell line. Passaging attempts in cell lines of mammalian origin failed for all three discovered rhabdoviruses. [ABSTRACT FROM AUTHOR]

Ticks are the main arthropod vector of pathogens to humans and livestock in the British Isles. Despite their role as a vector of disease, many aspects of tick biology, ecology, and microbial association are poorly understood. To address this, we investigated the composition of the microbiome of adult and nymphal Ixodes ricinus ticks. The ticks were collected on a dairy farm in Southwest England and RNA extracted for whole genome sequencing. Sequences were detected from a range of microorganisms, particularly tick-associated viruses, bacteria, and nematodes. A majority of the viruses were attributed to phlebo-like and nairo-like virus groups, demonstrating a high degree of homology with the sequences present in I. ricinus from mainland Europe. A virus sharing a high sequence identity with Chimay rhabdovirus, previously identified in ticks from Belgium, was detected. Further investigations of I. ricinus ticks collected from additional sites in England and Wales also identified Chimay rhabdovirus viral RNA with varying prevalence in all tick populations. This suggests that Chimay rhabdovirus has a wide distribution and highlights the need for an extended exploration of the tick microbiome in the United Kingdom (UK). [ABSTRACT FROM AUTHOR]

Objective: Ticks are arthropods found in many parts of the world that parasitize vertebrates, including livestock, wild animals, and humans. The distribution of tick in animals is an issue that needs to be taken into consideration as it also causes great economic losses. This study aims to detect ticks in the cattle population raised in Çankırı (province, Türkiye). Methods: In this study, tick samples were collected from 150 cattle between June and October 2022. Tick samples (n=1196) were identified morphologically using a stereomicroscope. Results: All collected ticks were adults, of these 516 (43.1%) were males and 680 (56.9%) females. With 845 (70.7%) specimens Haemaphysalis punctata was the most prevalent species, followed by Dermacentor marginatus 120 (10%), R. sanguineus 113 (9.4%). The least common tick type was Ixodes ricinus with 4 (0.3%), followed by Rhipicephalus bursa 9 (0.8%), and Hyalomma marginatum 9 (0.8%). There was a significant difference in terms of gender distribution between tick species (p<0.001), which was most pronounced in the case of R. sanguineus where 75% of the specimens were females and 25% males. R. sanguineus was the most prevalent species in Cardaklı and Yakalı; D. marginatus in Budakpınar and Soğluk; while Hae. puncatata in Eyüpözü, Susuz, Kızılıbrık, Ilıpınar, Hüyük, and Bozkuş District. The most common ticks in Susuz and Kızılıbrık belonged to the genus Haemaphysalis. Conclusion: This study contributed to health and economy by investigating the tick population in cattle. The distribution of ticks varies according to regions, climates and livestock activities. Since such studies have not been conducted in Çankırı before, we did not have the opportunity to compare with other studies. Prioritizing cattle has helped define the tick population in this region where animal husbandry is developed. By examining ticks, data was gained regionally in the fight against tick-borne diseases. In the study, the distribution of ticks between the Black Sea and Central Anatolia regions was examined and regional data was also obtained. [ABSTRACT FROM AUTHOR]

Background: Lyme borreliosis is the most common tick-borne disease in Europe and is often caused by Borrelia afzelii, which is transmitted by Ixodes ricinus ticks. The prevalence and abundance of infected ticks fluctuate in time and space, influencing human infection risk. Rodents are reservoir hosts for B. afzelii and important feeding hosts for larval ticks. In the study reported here, we examined how variation in rodent abundance is associated with B. afzelii infection prevalence in ticks, the density of nymphs (DON) and the density of infected nymphs (DIN) in the following year. We further analysed the relationships between the abundance of infected rodents and nymphal infection prevalence (NIP) and DIN. Methods: We conducted a study that combined experimental and observational approaches on 15 islands (10 small islands and 5 large islands) in Finland. On all of the islands, ticks and rodents were monitored and sampled during the summer of 2019, with the monitoring of tick abundance and sampling continuing into the spring of 2020. On five of the 10 small islands, captured rodents were removed from the island ("removal" islands), and on the other five small islands, captured rodents were released back to the trapping site after marking and sampling ("control" islands). On the five large islands, captured rodents were released back to the trapping site after marking and sampling. The presence of B. afzelii from nymph and rodent samples was examined. Results: The results of the experimental study showed that neither treatment (removal), rodent abundance index nor abundance index of infected rodents in 2019 was associated with DON, NIP or DIN in 2020. Based on data from the observational study, the NIP in 2020 decreased with increasing rodent abundance index and abundance index of infected rodents in 2019. However, the DIN in 2020 was not associated with the rodent abundance index or the abundance index of infected rodents in 2019. In addition, in the observational study, DON in 2020 increased with increasing rodent abundance index. Conclusions: Our results suggest that low rodent abundance during the tick activity period is not sufficient for reducing the disease hazard and, hence, rodent removal may not be a feasible control measure in natural ecosystems. [ABSTRACT FROM AUTHOR]

The tick-borne encephalitis (TBE) virus has been confirmed by molecular analysis in the Caucasus region for the first time. The virus obtained from a tick Ixodes ricinus ex Caspian green lizard belongs to the Zausaev strain of the Siberian subtype (not to a strain of the European subtype highly distributed in the territories adjacent to the Caucasus). This unusual record indicates the need to study the role of lizards in the circulation of natural focal infections. [ABSTRACT FROM AUTHOR]

Birds play a crucial role in disseminating ticks that carry pathogens of high veterinary-medical importance. The aim of this study was to analyze data of a long-term tick collection from birds at a single stop-over site in Central Europe, Hungary. Over eight years (2015–2022) 5833 ticks (ten species) were collected from 2395 tick-infested birds. The most abundant species were Ixodes ricinus (n = 3971) and Haemaphysalis concinna (n = 1706). Ixodes ricinus nymphs and larvae were the most frequently occurring on resident and short-distance migratory birds with forest habitat but Ha. concinna was the most abundant species on reed-associated, long-distance migrants. Haemaphysalis concinna occurred mostly on birds feeding above the ground level, while I. ricinus predominated on ground feeding birds. Infestation with I. ricinus nymphs always peaked in the first half of the year, in contrast to larvae which were more abundant on avian hosts in the autumn. At the same time, Ha. concinna larvae and nymphs had their peak numbers in the summer. This is the first long-term study on the tick infestation of birds in Central Europe. The study shows that, migration distance, habitat type, and typical feeding level of birds, as well as characteristics of tick life cycle are all key factors in the role of birds as tick disseminators. It was revealed that Savi's Warbler (Locustella luscinioides) is the most frequent hosts of Ha. concinna in Central Hungary. [ABSTRACT FROM AUTHOR]

Background: Changing geographical and seasonal activity patterns of ticks may increase the risk of tick infestation and tick-borne pathogen (TBP) transmission for both humans and animals. Methods: To estimate TBP exposure of dogs and cats, 3000 female I. ricinus from these hosts were investigated for Anaplasma phagocytophilum and Borrelia species. Results: qPCR inhibition, which was observed for ticks of all engorgement stages but not questing ticks, was eliminated at a template volume of 2 µl. In ticks from dogs, A. phagocytophilum and Borrelia spp. prevalence amounted to 19.0% (285/1500) and 28.5% (427/1500), respectively, while ticks from cats showed significantly higher values of 30.9% (464/1500) and 55.1% (827/1500). Accordingly, the coinfection rate with both A. phagocytophilum and Borrelia spp. was significantly higher in ticks from cats (17.5%, 262/1500) than dogs (6.9%, 104/1500). Borrelia prevalence significantly decreased with increasing engorgement duration in ticks from both host species, whereas A. phagocytophilum prevalence decreased only in ticks from dogs. While A. phagocytophilum copy numbers in positive ticks did not change significantly over the time of engorgement, those of Borrelia decreased initially in dog ticks. In ticks from cats, copy numbers of neither A. phagocytophilum nor Borrelia spp. were affected by engorgement. Borrelia species differentiation was successful in 29.1% (365/1254) of qPCR-positive ticks. The most frequently detected species in ticks from dogs were B. afzelii (39.3% of successfully differentiated infections; 70/178), B. miyamotoi (16.3%; 29/178), and B. valaisiana (15.7%; 28/178), while B. afzelii (40.1%; 91/227), B. spielmanii (21.6%; 49/227), and B. miyamotoi (14.1%; 32/227) occurred most frequently in ticks from cats. Conclusions: The differences in pathogen prevalence and Borrelia species distribution between ticks collected from dogs and cats may result from differences in habitat overlap with TBP reservoir hosts. The declining prevalence of A. phagocytophilum with increasing engorgement duration, without a decrease in copy numbers, could indicate transmission to dogs over the time of attachment. The fact that this was not observed in ticks from cats may indicate less efficient transmission. In conclusion, the high prevalence of A. phagocytophilum and Borrelia spp. in ticks collected from dogs and cats underlines the need for effective acaricide tick control to protect both animals and humans from associated health risks. [ABSTRACT FROM AUTHOR]

The structure and biochemical properties of protease inhibitors from the thyropin family are poorly understood in parasites and pathogens. Here, we introduce a novel family member, Ir-thyropin (IrThy), which is secreted in the saliva of Ixodes ricinus ticks, vectors of Lyme borreliosis and tick-borne encephalitis. The IrThy molecule consists of two consecutive thyroglobulin type-1 (Tg1) domains with an unusual disulfide pattern. Recombinant IrThy was found to inhibit human host-derived cathepsin proteases with a high specificity for cathepsins V, K, and L among a wide range of screened cathepsins exhibiting diverse endo- and exopeptidase activities. Both Tg1 domains displayed inhibitory activities, but with distinct specificity profiles. We determined the spatial structure of one of the Tg1 domains by solution NMR spectroscopy and described its reactive center to elucidate the unique inhibitory specificity. Furthermore, we found that the inhibitory potency of IrThy was modulated in a complex manner by various glycosaminoglycans from host tissues. IrThy was additionally regulated by pH and proteolytic degradation. This study provides a comprehensive structure–function characterization of IrThy—the first investigated thyropin of parasite origin—and suggests its potential role in host–parasite interactions at the tick bite site. [ABSTRACT FROM AUTHOR]

Background: The ornate dog tick Dermacentor reticulatus is second only to the hard tick Ixodes ricinus in terms of importance as a vector of infectious organisms, especially of Babesia canis, the agent of canine babesiosis. Both the geographical range and local densities of D. reticulatus are steadily increasing in many regions of Europe. In the present study, we tested the hypothesis that D. reticulatus possesses an efficient breeding strategy that allows for a rapid increase in tick numbers and densities through the formation of mixed-sex clusters/aggregations while questing in the environment. Methods: An observational study was carried out in the spring of 2023, at three sites in two regions in Central and North-Eastern Poland, both characterised by high tick densities. At each site, a 400-m-long transect was inspected for questing ticks. All noted ticks were collected, and tick numbers and sexes per stem were recorded. Differences in tick distribution by site and sex were analysed statistically. Results: A total of 371 D. reticulatus (219 females, 152 males) ticks were collected from 270 grass stems over a combined 1200 m of transect. The majority of grass stems (74.4%) were occupied by just a single individual, with two-tick clusters the second most common category. The maximum number of D. reticulatus individuals observed on a single grass stem was six. Mixed-sex clusters were significantly more common than single-sex clusters at all three sites. With study sites combined, mixed-sex clusters accounted for 17.4% (95% confidence limit [95% CL] 13.9–21.6%) of observations, while for multiple males and multiple females, the values were 2.6% (95% CL: 1.4–4.7%) and 5.6% (95% CL: 3.7–8.3%), respectively. Conclusions: Mixed-sex clusters of D. reticulatus ticks were significantly more common than single-sex clusters, which we hypothesise reflects an efficient, likely pheromone-mediated breeding strategy of this expansive tick species. [ABSTRACT FROM AUTHOR]

The Baltic states are the region in Europe where tick-borne encephalitis (TBE) is most endemic. The highest notification rate of TBE cases is reported in Lithuania, where the incidence of TBE has significantly increased since 1992. A recent study reported 0.4% prevalence of TBE virus (TBEV) in the two most common tick species distributed in Lithuania, Ixodes ricinus and Dermacentor reticulatus, with the existence of endemic foci confirmed in seven out of Lithuania's ten counties. However, until now, no comprehensive data on molecular characterisation and phylogenetic analysis have been available for the circulating TBEV strains. The aim of this study was to analyse TBEV strains derived from I. ricinus and D. reticulatus ticks collected from Lithuania and provide a genotypic characterisation of viruses based on sequence analysis of partial E protein and NS3 genes. The 54 nucleotide sequences obtained were compared with 81 TBEV strains selected from the NCBI database. Phylogenetic analysis of the partial E and NS3 gene sequences derived from 34 Lithuanian TBEV isolates revealed that these were specific to Lithuania, and all belonged to the European subtype, with a maximum identity to the Neudoerfl reference strain (GenBank accession no. U27495) of 98.7% and 97.4%, respectively. The TBEV strains showed significant regional genetic diversity. The detected TBEV genotypes were not specific to the tick species. However, genetic differences were observed between strains from different locations, while strains from the same location showed a high similarity. [ABSTRACT FROM AUTHOR]

Background: Many pathogens and parasites can infect multiple host species, and the competence of different hosts as pathogen reservoirs is key to understanding their epidemiology. Small mammals are important hosts for the instar stages of Ixodes ricinus ticks, the principal vector of Lyme disease in Europe. Small mammals also act as reservoirs of Borrelia afzelii, the most common genospecies of the Borrelia burgdorferi sensu lato (s.l.) spirochetes causing Lyme disease in Europe. However, we lack quantitative estimates on whether different small mammal species are equally suitable hosts for feeding I. ricinus and whether they show differences in pathogen transmission from host to tick. Methods: Here, we analysed the feeding success and prevalence of B. burgdorferi s.l. infections in 12,987 instar I. ricinus found on captured small mammals with known infection status in Norway (2018–2022). Results: We found that larvae were more likely to acquire a blood meal from common shrews (Sorex araneus, 46%) compared to bank voles (Myodes glareolus, 36%) and wood mice (Apodemus sylvaticus, 31%). Nymphs tended to be more likely to acquire a blood meal from wood mice (66%) compared to bank voles (54%). Common shrews harboured few nymphs (n=19). Furthermore, we found that larvae feeding on infected bank voles (11%) were more likely to be infected with B. burgdorferi s.l. than larvae on infected common shrews (7%) or wood mice (4%). Conclusions: Our study provides quantitative evidence of differences in suitability for the instar stages of I. ricinus across taxa of small mammals and highlights how even known small mammal host species can differ in their ability to feed ticks and infect larval ticks with the pathogen causing Lyme disease. [ABSTRACT FROM AUTHOR]

Background: The European hedgehog (Erinaceus europaeus) is known for high levels of ectoparasitism that not only represents a health risk for the animals themselves, but also for pet animals and humans as hedgehogs are frequently taken into human care. In the present study, patterns of ectoparasite infestation were assessed in hedgehogs taken into care at northern German animal rehabilitation centres. Methods: Ectoparasites (ticks, fleas and mites) of 498 hedgehogs were collected over a period of 3 years from July 2018 to May 2021. Species were identified based on morphological characteristics and also via amplification and sequencing of the partial cytochrome c oxidase subunit 2 (COX-2) gene for fleas of the family Ceratophyllidae. Seasonal changes in infestation patterns as well as correlations with animal age, body weight and health status were assessed using generalised linear models. Results: Infestation with ticks, fleas and mites occurred throughout the year. Overall, 86.5% (431/498) of the examined hedgehogs were infested with ticks, 91.4% (455/498) with fleas and 17.7% (88/498) with mites.Ixodes ricinus and Ixodes hexagonus/Ixodes canisuga were the most common tick species detected, with the additional occurrence of one Ixodes frontalis. Significant seasonal changes were observed for I. ricinus, but not for I. hexagonus/I. canisuga. Additionally, I. ricinus nymph prevalence declined significantly as of 2020, probably as a consequence of the climate change-related drought as of 2018. In hedgehogs with flea infestations, Archaeopsylla erinacei, Ceratophyllus sciurorum, Nosopsyllus fasciatus and Ctenocephalides felis were identified. In all cases of mite infestation, Caparinia tripilis was detected, in addition to specimens of the family Macronyssidae and free-living mites of the family Acaridae. Statistical analyses showed correlations regarding the factors month, year, body weight and age, but no correlation was evident regarding the health status of the animals. Conclusions: With a detected infestation rate of 98.6%, almost all of the examined hedgehogs were infested with at least one ectoparasite species. The seasonal activity patterns of the different ectoparasite species together with the complex annual cycle of hedgehogs lead to different seasonal patterns in ectoparasite prevalence and infestation intensities. Due to the risk of transmission of zoonotic pathogens as well as the possible negative impact on the host itself, hedgehogs should be treated against ectoparasites when taken into care facilities. [ABSTRACT FROM AUTHOR]

There is mounting evidence of increases in tick (Acari: Ixodidae) contacts in Finland during the past few decades, highlighted by increases in the incidence of Lyme borreliosis and tick-borne encephalitis (TBE). While nationwide field studies to map distributions of ticks are not feasible, crowdsourcing provides a comprehensive method with which to assess large-scale changes in tick contact areas. Here, we assess changes in tick contact areas in Finland between 1958 and 2021 using three different nationwide crowdsourced data sets. The data revealed vast increases in tick contact areas, with ticks estimated to be contacted locally approximately 400 km further north in western and approximately 100 km further north in eastern Finland in 2021 than 1958. Tick contact rates appeared to be highest along the coastline and on the shores of large lakes, possibly indicating higher tick abundance therein. In general, tick observations per inhabitant increased from 2015 to 2021. Tick contact areas have expanded in Finland over the past 60 years. It appears that taiga ticks (Ixodes persulcatus) are behind most of the northwards shifts in tick contact areas, with Ixodes ricinus contributing mostly to new contact areas in the south. While ticks are now present in most of Finland, there are still areas where tick abundance is low and/or establishment not possible, mainly in northern Finland. [ABSTRACT FROM AUTHOR]

Heterogeneous aggregation of parasites between individual hosts is common and regarded as an important factor in understanding transmission dynamics of vector-borne diseases. Lyme disease is vectored by generalist tick species, yet we have a limited understanding of how individual heterogeneities within small mammal host populations affect the aggregation of ticks and likelihood of infection. Male hosts often have higher parasite and infection levels than females, but whether this is linked to sexual body size dimorphism remains uncertain. Here, we analysed how host species, sex, and body mass influenced Ixodes ricinus tick infestations and the infection prevalence of Borrelia burgdorferi sensu lato (s.l.) in three species of small mammals involved in the enzootic transmission cycle of Lyme disease in Norway from 2018 to 2022. Larval and nymphal ticks were found on 98% and 34% of all individual hosts, respectively. In bank voles and wood mice, both larval and nymphal tick infestation and infection probability increased with body mass, and it increased more with mass for males than for females. Tick infestation in the common shrew increased with body mass and was higher in males, while pathogen infection was higher in females. Sex-biases in infestation did not correspond with level of sexual body mass dimorphism across species. This study contributes to our understanding of how individual heterogeneity among small mammalian hosts influences I. ricinus tick aggregation and prevalence of B. burgdorferi s.l. at northern latitudes. [ABSTRACT FROM AUTHOR]

People may be exposed to questing Ixodes ricinus ticks in urban settings, e.g. residential gardens. Little is known about the garden characteristics that support a tick population. To determine which features in and around residential gardens support or limit the occurrence and abundance of questing I. ricinus ticks, we sampled them in residential gardens in the Braunschweig region that differed in various intrinsic and extrinsic parameters. We recorded the number of questing nymphal and adult ticks on transects, and by using mixed-effects generalized linear regression models, we related their occurrence and abundance to garden characteristics, meteorological covariates, and landscape features in the vicinity. We detected questing I.ricinus ticks in about 90% of the 103 surveyed gardens. Our occurrence model (marginal R2 = 0.31) predicted the highest probability of questing ticks on transects with hedges or groundcover in gardens, which are located in neighborhoods with large proportions of forest. The abundance of questing ticks was similarly influenced. We conclude that I.ricinus ticks are frequent in residential gardens in Northern Germany and likely associated with intrinsic garden characteristics on a small scale, such as hedges, as well as extrinsic factors on a local scale, such as the proportion of nearby woodland. [ABSTRACT FROM AUTHOR]

Tick‐borne encephalitis virus (TBEV) is found in Ixodes ricinus ticks throughout the area where viable tick populations exist. In Norway, TBEV is found in I. ricinus from the south coast until Brønnøy municipality in Nordland County and the range of the vector is expanding due to changes in climate, vegetation, host animals and environmental conditions. TBEV might thus have the potential to establish in new areas when I. ricinus expand its geographical distribution. At present, there is little knowledge on the status of the virus in high‐altitude areas of inland regions in Norway. It has previously been indicated that reindeer may be an important sentinel species and indicator of the spread of ticks and TBEV in high‐altitude regions. In this study, 408 semi‐domesticated Eurasian tundra reindeer (Rangifer tarandus tarandus) from eight herds, from Tana in Troms and Finnmark County in northern Norway to Filefjell in Innlandet and Viken Counties in southern Norway, were screened for TBEV antibodies using a commercial enzyme‐linked immunosorbent assay (ELISA). We found 16 TBEV reactive reindeer samples by ELISA; however, these results could not be confirmed by the serum neutralization test (SNT). This could indicate that a flavivirusand not necessarily TBEV, may be circulating among Norwegian semi‐domesticated reindeer. The results also indicate that TBEV was not enzootic in Norwegian semi‐domesticated reindeer in 2013–2015. This knowledge is important as an information base for future TBEV and flavivirus surveillance in Norway. [ABSTRACT FROM AUTHOR]

The article focuses on the first instance of Amblyomma varium ticks serving as hosts for Ixodiphagus wasps. It describes the parasitism process, including the emergence of wasps from the ticks. It is further reported that this discovery contributes to our understanding of the ecological interactions between parasitoid wasps and ticks.

The tick-borne encephalitis virus (TBEV) is transmitted to humans through tick bites. In recent years, the appearance of the Siberian subtype of TBEV in Ixodes ricinus in Finland, together with deaths from the normally mild European subtype in the same country, have raised concerns about a possible spread of virulent variants of TBEV in Western Europe. Thus, there is a need to monitor the spread of strains, particularly of the European and Siberian subtypes. In this study, we develop a new real-time PCR method targeting Siberian and European subtypes of TBEV. The primers amplify a 176 bp fragment of the E gene, which is suitable for subsequent strain identification by Sanger sequencing. This study pioneers a new approach to primer design where the melting temperature (Tm) of primers annealed to representative mismatched target sequences is empirically determined and used to guide improvements in primer sequence. This allowed the range of TBEV strains detected to be extended to cover most European and Siberian strains tested, in addition to a strain of the Far-Eastern subtype. The limit of detection was 10–100 DNA copies per reaction and amplification efficiency varied between 83% and 94%, depending on the TBEV strain. Experimental determination of primer Tm proved to be a fruitful approach and will be a useful tool for future primer design and diagnostics. [ABSTRACT FROM AUTHOR]

Background: Ixodes ricinus ticks are infected with a large diversity of vertically and horizontally transmitted symbionts. While horizontally transmitted symbionts rely on a vertebrate host for their transmission, vertically transmitted symbionts rely more on the survival of their invertebrate host for transmission. We therefore hypothesized horizontally transmitted symbionts to be associated with increased tick activity to increase host contact rate and vertically transmitted symbionts to be associated with higher tick weight and lipid fraction to promote tick survival. Methods: We used a behavioural assay to record the questing activity of I. ricinus ticks. In addition, we measured weight and lipid fraction and determined the presence of ten symbiont species in these ticks using qPCR, of which six were vertically transmitted and four horizontally transmitted. Results: Vertically transmitted symbionts (e.g. Midichloria mitochondrii) were associated with an increase in tick weight, whereas horizontally transmitted symbionts (e.g. Borrelia burgdorferi sensu lato) were often associated with lower weight and lipid fraction of ticks. Moreover, horizontally transmitted symbionts (e.g. B. burgdorferi s.l.) were associated with increased tick activity, which may benefit pathogen transmission and increases tick-borne disease hazard. Conclusions: Our study shows that horizontally and vertically transmitted symbionts differentially influence the behaviour and physiology of I. ricinus and warrants future research to study the underlying mechanisms and effects on transmission dynamics of tick-borne pathogens. [ABSTRACT FROM AUTHOR]

Tick-borne pathogens are responsible for many vector-borne diseases in Europe, causing important problems for human and animal health. The composition of viral communities in ticks and their interactions with pathogens is little understood, especially in Eastern Europe, an area that represents a major hub for animal-arthropod vectors exchanges. In this study, we used metatranscriptomics to characterize the virome of 2,753 Ixodes ricinus ticks collected from France and Romania, focusing on viruses that could potentially have implications for human or animal health. Among the great viral diversity of viruses identified, we reported a novel strain of Tribec virus, an important human pathogen that was found in Romanian ticks. We detected viruses belonging to the Phenuiviridae and Nairoviridae families close to human and animal pathogens, suggesting that these viruses could constitute novel arboviruses. We used luciferase immunoprecipitation system targeting external viral proteins of viruses identified among the Sedoreoviridae, Phenuiviridae, and Nairoviridae families to screen serum samples from small ruminants' exposed to tick bites. The results suggest that part (approximately 12%, 95% CI 9.1–16.2) of the small ruminant population from Danube Delta, was exposed to viruses related to bi- or tri-segmented nairoviruses, but cross-reactive viruses could not be confirmed with certainly. The strategy developed in this study serves as a key step in predicting potential new disease outbreaks and can be readily adapted to study other reservoirs, vectors, and interfaces involving susceptible hosts. [ABSTRACT FROM AUTHOR]

Background: Anaplasma phagocytophilum is a Gram-negative obligate intracellular bacterium that replicates in neutrophil granulocytes. It is transmitted by ticks of the Ixodes ricinus complex and causes febrile illness called granulocytic anaplasmosis primarily in humans, horses, dogs, sheep, cattle and goats. In comparison, clinically apparent disease has been described rarely in cats especially compared to dogs and horses. It is currently unknown whether cats are less susceptible to A. phagocytophilum or whether granulocytic anaplasmosis might be underdiagnosed in cats. Methods: To address this question, we examined clinical signs and laboratory findings in seven A. phagocytophilum infected cats from Germany and Switzerland. We then genetically characterized feline A. phagocytophilum strains and compared them to those from other hosts showing clinically apparent disease. For this purpose, ankA-based, groEL-based and multilocus sequence typing (MLST) were applied. Furthermore, the concordance between these typing methods was assessed. Results: Fever, lethargy and anorexia were the most common clinical signs in cats suffering from granulocytic anaplasmosis. The most frequent laboratory finding was thrombocytopenia. All three typing methods consistently indicated that the A. phagocytophilum strains found infecting cats are the same as those that cause disease in humans, dogs and horses. In general, the three typing methods applied exhibited high concordance. Conclusions: The genetic characterization of the feline A. phagocytophilum strains indicates that strain divergence is not the explanation for the fact that granulocytic anaplasmosis is much less frequently diagnosed in cats than in dogs and horses. Otherwise, it may be possible that cats are less susceptible to the same strains than dogs and horse are. However, due to the unspecific clinical signs, it should be considered that granulocytic anaplasmosis may be under-diagnosed in cats. [ABSTRACT FROM AUTHOR]

Background: Borrelia persica causes tick-borne relapsing fever in Israel, the eastern Mediterranean basin, and Asia. Relapsing fever is associated with severe illness and potentially death in humans and animals. Since B. persica infection has rarely been described in wild animals, the aim of this study was to evaluate the prevalence of infection with B. persica in wild carnivores in Israel. Methods: Spleen and blood clot samples from wild carnivores, which underwent necropsy, were tested for the presence of Borrelia DNA by real-time polymerase chain reaction (PCR). PCR products were sequenced, and the spirochete loads were quantified using a specific quantitative PCR (qPCR). Results: A total of 140 samples from 74 wild carnivores were analyzed for the presence of Borrelia DNA. Six out of the 74 (8.1%) animals were found positive for B. persica by PCR and sequencing of the flagellin B gene, of which 4/74 (5.4%) were also positive by PCR for the glycerophosphodiester phosphodiesterase (glpQ) gene. Positive samples were obtained from three European badgers, and one striped hyena, golden jackal, and red fox each. All B. persica-positive animals were young males (P < 0.0001). Quantifiable results were obtained from 3/5 spleen and 4/5 blood samples. The spirochete loads in the blood were significantly higher than those found in the spleen (P = 0.034). Conclusions: The prevalence of B. persica infection found in wild carnivores brought for necropsy was unexpectedly high, suggesting that this infection is widespread in some wild animal species in Israel. This is the first report of B. persica infection in the European badger and striped hyena. These carnivores have a wide geographical range of activity, and the results of this survey raise the possibility that they may serve as reservoir hosts for B. persica. [ABSTRACT FROM AUTHOR]

Background: Ticks carry microbes, some of which are pathogenic for humans and animals. To assess this One Health challenge, 342 ticks were collected from pet dogs and cats at 10 veterinary clinics in Finland as part of the European project "Protect Our Future Too". Methods: The tick species were identified, and ticks were screened with quantitative PCR (qPCR) for tick-borne pathogens, including Borrelia burgdorferi sensu lato, Borrelia miyamotoi, Ehrlichia canis, Anaplasma spp., Candidatus Neoehrlichia mikurensis, tick-borne encephalitis virus (TBEV), and Babesia spp. For comparison, a subset of tick DNA (20 qPCR-positive samples) was analysed with 16S next-generation sequencing (NGS). Results: Most ticks were Ixodes ricinus (289, 84.5%), followed by Ixodes persulcatus (51, 14.9%). One hybrid tick (I. ricinus/I. persulcatus, 0.3%) and one Rhipicephalus sanguineus tick (0.3%) were identified. We found one or more of the analysed pathogens in 17% (59/342) of the ticks. The most prevalent pathogen was B. burgdorferi s.l. (36, 10.5%), followed by Anaplasma phagocytophilum (12, 3.5%), B. miyamotoi (5, 1.5%), Babesia venatorum (4, 1.2%), and TBEV (1, 0.3%). Candidatus Neoehrlichia mikurensis DNA was amplified from three (0.9%) ticks. Ehrlichia canis was not detected. In the 16S NGS, six samples produced enough reads for the analysis. In these six samples, we confirmed all the positive qPCR findings of Borrelia spp. and Ca. N. mikurensis. Conclusions: The high prevalence of pathogenic microorganisms in the ticks of this study emphasizes the importance of awareness of ticks and tick-borne diseases and prevention. Furthermore, the results show that veterinary surveillance can facilitate early detection of tick-borne pathogens and new tick species and draw attention to possible co-infections that should be considered both in symptomatic humans and animals after tick bites. [ABSTRACT FROM AUTHOR]

Recently, increasing attention has been posed on the role of migrating birds in the spread of ticks and indirectly of tick-borne pathogens (TBPs). Despite, Italy is considered a bridge between continental Europe and North Africa and a necessary path to connect Mediterranean countries, few studies have been conducted on ticks collected from birds migrating through this country and associated TBPs. The aims of this research were to estimate the infestation burden and identify tick species feeding on migratory birds, and perform a molecular screening for TBPs. During autumn migration (2019–2020), birds were inspected for ticks in a ringing station located in north-east Italy. Ticks were identified and screened for tick-borne encephalitis virus (TBEV), Borrelia burgdorferi sensu latu, Rickettsia spp., Ehrlichia spp., Neoehrlichia spp., Anaplasma phagocytophilum, and Bartonella spp. Ixodes ticks (n = 209) were feeding on 2.6% of passerines (88/3411). Most of these (208/209) were Ixodes ricinus, except one Ixodes frontalis. Eight bird species were infested: common blackbird, redwing, brambling, song thrush, common chaffinch, European robin, water pipit, and coal tit. Rickettsiales showed a low prevalence, from 1.4% of Ehrlichia spp., 4.3% of A. phagocytophilum, up to 7.2% of Rickettsia spp.. Borrelia burgdorferi s.l. had the highest prevalence, 54.6%, and several zoonotic genotypes were identified: B. garinii, B. valaisiana, B. afzeli, B. burgdorferi sensu stricto, and B. miyamotoi. All specimens were negative for TBEV and Bartonella spp.. Although the tick burden was generally low, most of the vectors (>60%) were positive for at least one pathogen, highlighting the relevance of a continuous monitoring of migrating birds as potential sources of pathogen dispersal. [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]

[Display omitted] • German samples previously identified as Ixodes inopinatus represent Ixodes ricinus based on genomic data. • The mitochondrial genome is not sufficient for delineation of I. inopinatus and I. ricinus. • The German samples most likely do not represent I. ricinus / I. inopinatus hybrids. Ticks are important vectors of human and animal pathogens, but many questions remain unanswered regarding their taxonomy. Molecular sequencing methods have allowed research to start understanding the evolutionary history of even closely related tick species. Ixodes inopinatus is considered a sister species and highly similar to Ixodes ricinus , an important vector of many tick-borne pathogens in Europe, but identification between these species remains ambiguous with disagreement on the geographic extent of I. inopinatus. In 2018–2019, 1583 ticks were collected from breeding great tits (Parus major) in southern Germany, of which 45 were later morphologically identified as I. inopinatus. We aimed to confirm morphological identification using molecular tools. Utilizing two genetic markers (16S rRNA, TROSPA) and whole genome sequencing of specific ticks (n = 8), we were able to determine that German samples, morphologically identified as I. inopinatus , genetically represent I. ricinus regardless of previous morphological identification, and most likely are not I. ricinus / I. inopinatus hybrids. Further, our results showed that the entire mitochondrial genome, let alone singular mitochondrial genes (i.e., 16S), is unable to distinguish between I. ricinus and I. inopinatus. Our results suggest that I. inopinatus is geographically isolated as a species (northern Africa and potentially southern Spain and Portugal) and brings into question whether I. inopinatus exists in central Europe. Our results highlight the probable existence of I. inopinatus and the power of utilizing genomic data in answering questions regarding tick taxonomy. [ABSTRACT FROM AUTHOR]

Wild animals play an important role as amplifiers and/or reservoir hosts in the ecology of many ticks and tick-borne pathogens affecting livestock and humans. In this study, which was carried out in the Hatay province, the southernmost region of Anatolia, a total of 362 (210°C, 146 °C, 6N) tick specimens were collected from 18 wild animals from 7 species, including white stork (Ciconia ciconia) (n = 1), roe deer (Capreolus capreolus) (n = 5), badger (Meles meles) (n = 2), jackal (Canis aureus) (n = 3), red fox (Vulpes vulpes) (n = 5), hare (Lepus europaeus) (n = 1), and wild goat (Capra aegagrus) (n = 1), which were obtained between 2014 and 2022. The collected ticks were identified according to morphological criteria at the level of species as Amblyomma lepidum, Dermacentor marginatus, Haemaphysalis erinacei, Haemaphysalis inermis, Haemaphysalis kopetdaghica, Ixodes gibbosus, Ixodes kaiseri, Ixodes ricinus, Rhipicephalus kohlsi, Rhipicephalus rossicus, and Rhipicephalus turanicus. With this study, A. lepidum was reported for the first time in Türkiye, while R. rossicus detected in roe deer was reported for the first time in wild animals, and the lesser-known/rare tick H. kopetdaghica was reported for the second time in wild goats where it was previously reported. [ABSTRACT FROM AUTHOR]

The documentation of endogenous viral elements (EVEs; virus-derived genetic material integrated into the genome of a nonviral host) has offered insights into how arthropods respond to viral infection via RNA interference pathways. Small non-coding RNAs derived from EVE loci serve to direct RNAi pathways in limiting replication and infection from cognate viruses, thus benefiting the host's fitness and, potentially, vectorial capacity. Here we use informatic approaches to analyze nine available genome sequences of hard ticks (Acari: Ixodidae; Rhipicephalus sanguineus, R. microplus, R. annulatus, Ixodes ricinus, I. persulcatus, I. scapularis, Hyalomma asiaticum, Haemaphysalis longicornis, and Dermacentor silvarum) to identify endogenous viral elements and to illustrate the shared ancestry of all elements identified. Our results highlight a broad diversity of viral taxa as having given rise to 1234 identified EVEs in ticks, with Mononegavirales (specifically Rhabdoviridae) well-represented in this subset of hard ticks. Further investigation revealed extensive adintovirus integrations in several Ixodes species, the prevalence of Bunyavirales EVEs (notably not observed in mosquitoes), and the presence of several elements similar to known emerging human and veterinary pathogens. These results will inform subsequent work on current and past associations with tick species with regard to the viruses from which their "viral fossils" are derived and may serve as a reference for quality control of various tick-omics data that may suffer from misidentification of EVEs as viral genetic material. [ABSTRACT FROM AUTHOR]

Background: The ticks Ixodes ricinus and Dermacentor reticulatus are two of the most important vectors in Europe. Climate niche modelling has been used in many studies to attempt to explain their distribution and to predict changes under a range of climate change scenarios. The aim of this study was to assess the ability of different climate niche modelling approaches to explain the known distribution of I. ricinus and D. reticulatus in Europe. Methods: A series of climate niche models, using different combinations of input data, were constructed and assessed. Species occurrence records obtained from systematic literature searches and Global Biodiversity Information Facility data were thinned to different degrees to remove sampling spatial bias. Four sources of climate data were used: bioclimatic variables, WorldClim, TerraClimate and MODIS satellite-derived data. Eight different model training extents were examined and three modelling frameworks were used: maximum entropy, generalised additive models and random forest models. The results were validated through internal cross-validation, comparison with an external independent dataset and expert opinion. Results: The performance metrics and predictive ability of the different modelling approaches varied significantly within and between each species. Different combinations were better able to define the distribution of each of the two species. However, no single approach was considered fully able to capture the known distribution of the species. When considering the mean of the performance metrics of internal and external validation, 24 models for I. ricinus and 11 models for D. reticulatus of the 96 constructed were considered adequate according to the following criteria: area under the receiver-operating characteristic curve > 0.7; true skill statistic > 0.4; Miller's calibration slope 0.25 above or below 1; Boyce index > 0.9; omission rate < 0.15. Conclusions: This comprehensive analysis suggests that there is no single 'best practice' climate modelling approach to account for the distribution of these tick species. This has important implications for attempts to predict climate-mediated impacts on future tick distribution. It is suggested here that climate variables alone are not sufficient; habitat type, host availability and anthropogenic impacts, not included in current modelling approaches, could contribute to determining tick presence or absence at the local or regional scale. [ABSTRACT FROM AUTHOR]

Background: Alpha-Gal syndrome (AGS) is a tick-borne food allergy caused by IgE antibodies against the glycan galactose-alpha-1,3-galactose (α-Gal) present in glycoproteins and glycolipids from mammalian meat. To advance in the diagnosis and treatment of AGS, further research is needed to unravel the molecular and immune mechanisms underlying this syndrome. The objective of this study is the characterization of tick salivary components and proteins with and without α-Gal modifications involved in modulating human immune response against this carbohydrate. Methods: Protein and α-Gal content were determined in tick saliva components, and proteins were identified by proteomics analysis of tick saliva fractions. Pathophysiological changes were recorded in the zebrafish (Danio rerio) model after exposure to distinct Ixodes ricinus tick salivary components. Serum samples were collected from zebrafish at day 8 of exposure to determine anti-α-Gal, anti-glycan, and anti-tick saliva protein IgM antibody titers by enzyme-linked immunosorbent assay (ELISA). Results: Zebrafish treated with tick saliva and saliva protein fractions combined with non-protein fractions demonstrated significantly higher incidence of hemorrhagic type allergic reactions, abnormal behavioral patterns, or mortality when compared to the phosphate-buffered saline (PBS)-treated control group. The main tick salivary proteins identified in these fractions with possible functional implication in AGS were the secreted protein B7P208-salivary antigen p23 and metalloproteases. Anti-α-Gal and anti-tick salivary gland IgM antibody titers were significantly higher in distinct saliva protein fractions and deglycosylated saliva group when compared with PBS-treated controls. Anti-glycan antibodies showed group-related profiles. Conclusions: Results support the hypothesis that tick salivary biomolecules with and without α-Gal modifications are involved in modulating immune response against this carbohydrate. [ABSTRACT FROM AUTHOR]

Borrelia burgdorferi sensu lato (Bbsl) is spirochetes transmitted by ticks and known to cause Lyme disease. Chlamydia-like organisms (CLOs) comprise a large group of bacteria that can lead to serious health disorders, including miscarriage. Recently, CLOs have been found in ticks and patient skin biopsies. Due to the involvement of multiple potential vectors in the spread of these pathogens, the objective of this study was to confirm the presence of both organisms in the early developmental stages of selected vectors. Three potential vectors, Ixodes ricinus larvae, Culex pipiens larvae, and winged (unfed) adults of Lipoptena cervi, were collected in the Czech Republic in years 2019–2020. The presence of Bbsl and panchlamydial DNA was detected by PCR and positive samples were further analyzed by Sanger sequencing and phylogenetic tree construction. Bbsl DNA was proved in 1.5% (2/137) of I. ricinus larvae (identified as Borrelia afzelii and Borrelia garinii), in 1.7% (2/119) of C. pipiens larvae (both identified as B. garinii), and in 11% (3/27) of L. cervi (all identified as B. garinii). CLOs were identified in 0.7% (1/137) of I. ricinus larvae (Candidatus Protochlamydia) and in 7.4% (2/27) of L. cervi (unspecified genus), while C. pipiens larvae could not be evaluated (0%). This research represents the first investigation of the presence of CLOs in L. cervi. The detection of pathogen DNA in the early developmental stages of vectors suggests the potential for transgenerational transmission of Bbsl and CLOs in the selected vectors, although at a low rate. [ABSTRACT FROM AUTHOR]

Simple Summary: Ixodes species are the main vectors of bacteria and piroplasm for different vertebrate hosts. Research on these unexplored concerns has been neglected in different regions including Pakistan. Recently, we molecularly characterized Ixodes kashmiricus ticks and associated Rickettsia spp.; however, the cox1 sequence and associated Theileria spp. and Anaplasma spp. for this tick are unknown. This study aimed to genetically identify I. kashmiricus based on the cox1 sequence and associated Theileria spp. and Anaplasma spp. A total of 352 ticks including adult females, nymphs and males were collected from small ruminants. The BLAST results and phylogenetic analysis of the cox1 sequence revealed a close resemblance with the Ixodes ricinus complex sequences. The 18S rDNA and 16S rDNA sequences showed maximum identity with Theileria cf. sinensis or Theileria sinensis and Anaplasma capra, respectively, and they phylogenetically clustered with the same species. This is the first report on the cox1 sequence of the I. kashmiricus tick, new locality records, and associated T. sinensis-like and A. capra. In order to determine the epidemiology of Ixodes ticks and their related pathogens, a widespread tick investigation is required. Ixodes ticks transmit Theileria and Anaplasma species to a wide range of animals. The spreading of ticks and tick-borne pathogens has been attributed to transhumant herds, and research on these uninvestigated issues has been neglected in many countries, including Pakistan. Recently, we used internal transcribed spacer (ITS) and 16S ribosomal DNA partial sequences to genetically characterize Ixodes kashmiricus ticks and their associated Rickettsia spp. However, the data on its cox1 sequence and associated Theileria spp. and Anaplasma spp. are missing. This study aimed to genetically characterize I. kashmiricus based on the cox1 sequence and their associated Theileria spp. and Anaplasma spp. The I. kashmiricus ticks were collected from small ruminants: sheep (Ovis aries) and goats (Capra hircus) of transhumant herds in district Shangla, Dir Upper and Chitral, Khyber Pakhtunkhwa (KP), Pakistan. Out of 129 examined hosts, 94 (72.87%) (56 sheep and 38 goats) were infested by 352 ticks, including adult females (175; 49.7%) followed by nymphs (115; 32.7%) and males (62; 17.6%). For molecular analyses, 121 ticks were subjected to DNA isolation and PCR for the amplification of the cox1 sequence for I. kashmiricus, 18S rDNA for Theileria spp. and 16S rDNA sequences for Anaplasma spp. The obtained cox1 sequence showed 89.29%, 88.78%, and 88.71% identity with Ixodes scapularis, Ixodes gibbosus, and Ixodes apronophorus, respectively. Phylogenetically, the present cox1 sequence clustered with the Ixodes ricinus complex. Additionally, the 18S rDNA sequence showed 98.11% maximum identity with Theileria cf. sinensis and 97.99% identity with Theileria sinensis. Phylogenetically, Theileria spp. clustered with the T. cf. sinensis and T. sinensis. In the case of Anaplasma spp., the 16S rDNA sequence showed 100% identity with Anaplasma capra and phylogenetically clustered with the A. capra. PCR-based DNA detection targeting the amplification of groEL and flaB sequences of Coxiella spp. and Borrelia spp., respectively, was unsuccessful. This is the first phylogenetic report based on cox1 and new locality records of I. kashmiricus, and the associated T. sinensis-like and A. capra. Significant tick surveillance studies are needed in order to determine the epidemiology of Ixodes ticks and their associated pathogens. [ABSTRACT FROM AUTHOR]

Background: Ixodes ricinus is an important vector of several pathogens, primarily in Europe. Recently, Ixodes inopinatus was described from Spain, Portugal, and North Africa and then reported from several European countries. In this study, a multiplex polymerase chain reaction (PCR) was developed to distinguish I. ricinus from I. inopinatus and used in the surveillance of I. inopinatus in Algeria (ALG) and three regions in the Czech Republic (CZ). Methods: A multiplex PCR on TROSPA and sequencing of several mitochondrial (16S rDNA, COI) and nuclear markers (TROSPA, ITS2, calreticulin) were used to differentiate these two species and for a subsequent phylogenetic analysis. Results: Sequencing of TROSPA, COI, and ITS2 separated these two species into two subclades, while 16S rDNA and calreticulin could not distinguish I. ricinus from I. inopinatus. Interestingly, 23 nucleotide positions in the TROSPA gene had consistently double peaks in a subset of ticks from CZ. Cloning of these PCR products led to a clear separation of I. ricinus and I. inopinatus indicating hybridization and introgression between these two tick taxa. Based on a multiplex PCR of TROSPA and analysis of sequences of TROSPA, COI, and ITS2, the majority of ticks in CZ were I. ricinus, no I. inopinatus ticks were found, and 10 specimens showed signs of hybridization. In contrast, most ticks in ALG were I. inopinatus, four ticks were I. ricinus, and no signs of hybridization and introgression were detected. Conclusions: We developed a multiplex PCR method based on the TROSPA gene to differentiate I. ricinus and I. inopinatus. We demonstrate the lack of evidence for the presence of I. inopinatus in Central Europe and propose that previous studies be re-examined. Mitochondrial markers are not suitable for distinguishing I. inopinatus from I. ricinus. Furthermore, our data indicate that I. inopinatus and I. ricinus can hybridize, and the hybrids can survive in Europe. [ABSTRACT FROM AUTHOR]

• An inactivated whole-virion TBEV vaccine candidate for dogs was developed. • The vaccine candidate was tested in mice and in dogs. • The vaccine was well tolerated and elicited production of neutralizing anti-TBEV antibodies. • Vaccination of mice provided complete protection against lethal TBEV infection. • Vaccination of dogs prevented the development of viremia after challenge infection. Tick-borne encephalitis (TBE) is a severe neuroinfection of humans. Dogs are also commonly infected with tick-borne encephalitis virus (TBEV). These infections are usually asymptomatic, but sometimes show clinical signs similar to those seen in humans and can be fatal. To date, there is no TBEV vaccine available for use in dogs. To address this need, a TBEV vaccine candidate for dogs based on inactivated whole virus antigen was developed. The safety, immunogenicity, and efficacy of the vaccine candidate were tested in mice as the preclinical model and in dogs as the target organism. The vaccine was well tolerated in both species and elicited the production of specific anti-TBEV antibodies with virus neutralising activity. Vaccination of mice provided complete protection against the development of fatal TBE. Immunisation of dogs prevented the development of viremia after challenge infection. Therefore, the developed vaccine candidate is promising to protect dogs from severe TBEV infections. [ABSTRACT FROM AUTHOR]