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The aim of the work was to determine the species diversity of the causative agents of Ixodidae tick-borne borrelioses in Ixodes persulcatus ticks in the Khabarovsk Territory.Materials and methods. During the epidemic season (April–October) 2017–2023, 4751 specimens of I. persulcatus Schulze, 1930, removed after attachment to humans and 418 ones collected from vegetation in the Khabarovsk Region, were studied. Ixodidae ticks were collected in the green areas of Khabarovsk city during the snowless season of 2021–2023, as well as in the territory of the Khabarovsk Region on the flag. DNA of the borrelia complex Borrelia burgdorferi sensu lato (s.l.) and B. miyamotoi was detected in ticks using real-time polymerase chain reaction (PCR). Differentiation of borrelia species in samples containing genetic material of B. burgdorferi s.l. was carried out in two stages. At the first stage, the presence of DNA from borrelia of the B. garinii s.l. group (B. garinii + B. bavariensis) and B. afzelii DNA was determined in the sample. At the second stage, positive samples of B. garinii s.l. were differentiated into B. garinii sensu stricto (s.s.) and B. bavariensis. Results and discussion. In engorged ticks, genetic material of B. burgdorferi s.l. was detected in 45.7 % of the cases, DNA of B. miyamotoi was identified in 7.2 % of samples. In ticks collected from vegetation, the DNA of B. burgdorferi s.l. was detected in 38.0 % of cases. Upon further study, the genetic material of B. afzelii and borrelia of the B. garinii s.l. group was identified in 47.2 % of cases for both pathogens. Within the group B. garinii s.l., DNA of B. bavariensis was detected in 18.6 %, B. garinii s.s. – in 8 % of samples, at the same time, mixed infection was noted in 53.3 % of cases. The infection rate with B. afzelii in I. persulcatus ticks turned out to be statistically significantly higher than that for B. garinii s.s. and B. bavariensis, thereat statistically significant differences in tick infection rates with B. garinii s.s. and B. bavariensis was not detected.

Abstract Background Urban green spaces are important for human health, but they may expose visitors to tick-borne diseases. This not only presents a potential public health challenge but also undermines the expected public health gains from urban green spaces. The aim of this study is to assess the public health risk of tick-borne diseases in an urban green space used for recreation in Stockholm, Sweden. Methods We used a mixed method approach identifying both the magnitude of the tick hazard and the extent of the human exposure to tick-borne diseases. At six entry points to an urban green space, we sampled ticks and documented microhabitat conditions from five randomly assigned 2 m × 2 m plots. Surrounding habitat data was analyzed using geographical information system (GIS). Nymphs and adult ticks were tested for Borrelia burgdorferi sensu lato and Anaplasma phagocytophilum using TaqMan qPCR. Positive B. burgdorferi (s.l.) ticks were further analyzed by nested PCR amplification and sequence analysis. Population census data and visitor count data were used to estimate the degree of human exposure to tick-borne diseases. To further understand the degree to which visitors get in contact with infected ticks we also conducted interviews with visitors to green spaces. Results High tick densities were commonly found in humid broadleaved forest with low field vegetation. High pathogen prevalence was significantly correlated with increasing proportions of artificial areas. Integrating the tick hazard with human exposure we found that the public health risk of tick-borne diseases was moderate to high at most of the studied entry points. Many of the visitors frequently used urban green spaces. Walking was the most common activity, but visitors also engaged in activities with higher risk for tick encounters. Individual protective measures were connected to specific recreational activities such as picking berries or mushrooms. Conclusions The number of visitors can be combined with tick inventory data and molecular analyses of pathogen prevalence to make crude estimations of the public health risk of tick-borne diseases in urban green spaces. The risk of encountering infected ticks is omnipresent during recreational activities in urban green spaces, highlighting the need for public health campaigns to reduce the risk of tick-borne diseases.

A 76-year-old woman infected with Yezo virus (YEZV) developed liver dysfunction and thrombocytopenia following a tick bite. Despite the severity of her elevated liver enzymes and reduced platelet counts, the patient's condition improved spontaneously without any specific treatment. To our knowledge, this represents the first documented case where the YEZV genome was detected simultaneously in a patient's serum and the tick (Ixodes persulcatus) that bit the patient. This dual detection not only supports the hypothesis that YEZV is a tick-borne pathogen but also underscores the importance of awareness and diagnostic readiness for emerging tick-borne diseases, particularly in regions where these ticks are prevalent.

Abstract 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.

Background. Infections transmitted to humans by the bites of ixodid ticks remain an urgent public health problem. In this work we explored the natural foci of tickborne infections located in the valley of the Chikoy River, which is a part of the buffer zone of the Baikal natural territory.The aim. To characterize the modern diversity and prevalence of tick-borne pathogens in the ecosystems of the valley of the Chikoy River (Trans-Baikal Territory, Russian Federation).Materials and methods. Thirteen sampling sites were located in typical biotopes throughout the Chikoy valley. In total 48 adult Ixodes persulcatus ticks, 1 female Haemaphysalis concinna tick and 38 specimens of small mammals were studied. All samples were tested for infection with seven tick-borne pathogens using multiplex real-time PCR.Results. No pathogens were detected in the H. concinna specimen. No R. sibirica and R. heilongjiangensis were detected both in ticks and in rodents. Among I. persulcatus, tick-borne encephalitis virus (TBEV), and the prevalence of Borrelia burgdorferi s. l. comprised 39.5 %, A. phagocytophilum – 16.7 %, B. miyamotoi – 8.3 % and Ehrlichia sp. – 2.1 %. Among infected ticks 6.2 % were co-infected with B. burgdorferi s. l. and A. phagocytophilum. Four rodent hosts of ticks and infections were identified: Myodes rufocanus (44.7 %), Apodemus peninsulae (39 %), Microtus oeconomus (13.2 %) and M. rutilus (2.6 %). Mean prevalence of B. burgdorferi s. l. in rodents comprised 39.5 %, B. miyamotoi – 28.9 %, Ehrlichia sp. – 21.1 % and A. phagocytophilum – 18.4 %. TBEV was detected in 5.3 % of rodents.Conclusion. At least five tick-borne pathogens circulate in the Chikoi River valley, i. e. TBEV, B. burgdorferi sensu lato, A. phagocytophilum, B. miyamotoi and Ehrlichia sp. The wide spread of I. persulcatus and abundance of competent rodent hosts of infections and ticks indicates that natural foci of tick-borne diseases are widely distributed in the Chikoi River valley.

Background. To date, the genetic variability of Rickettsia helvetica has not been sufficiently studied.The aim. To study the prevalence and genetic variability of R. helvetica in Ixodes spp. collected in Western Siberia and the Russian Far East.Materials and methods. Ixodes spp. collected from rodents in the Omsk province, Western Siberia (n = 280) and collected by flagging on Putyatin and Russky Islands in Primorsky Krai, Russian Far East (n = 482) were analyzed for the presence of Rickettsia spp. All positive samples were genotyped for the gltA gene fragment. For a number of R. helvetica samples, fragments of the 16S rRNA, ompA, ompB, sca4, htrA, and groEL genes and 23S–5S intergenic spacer were additionally sequenced.Results. Four Rickettsia species (R. helvetica, “Candidatus Rickettsia tarasevichiae”, “Candidatus Rickettsia uralica”, and “Candidatus Rickettsia mendelii”) were found. Of them, R. helvetica was identified in 72.2 % of Ixodes apronophorus and 18.8 % of Ixodes trianguliceps from the Omsk province and in single Ixodes persulcatus from the Omsk province and Putyatin Island. This is the first finding of Rickettsia spp. in I. apronophorus. All known R. helvetica sequences from this study and the GenBank database belonged to four well supported monopheletic groups forming genetic lineages I–IV. Lineage I included European isolates from Ixodes ricinus, Western Siberian isolates from I. persulcatus, and some sequences from I. apronophorus. All R. helvetica sequences from I. trianguliceps from the Omsk province and I. persulcatus from the Komi Republic and one sequence from I. apronophorus were assigned to lineage II. Most sequences from I. apronophorus formed lineage III; all known R. helvetica sequences from I. persulcatus from the Far East formed genetic lineage IV.Conclusion. The genetic heterogeneity of R. helvetica population was first demonstrated. Known isolates of R. helvetica are reliably assigned to four genetic lineages, but not in all cases the association of different lineages with a specific tick species or specific territory was observed.

Ticks are one of the most important vectors that can transmit pathogens to animals and human beings. This study investigated the dominant tick-borne bacteria carried by ticks and tick-borne infections in forestry populations in Arxan, Inner Mongolia, China. Ticks were collected by flagging from May 2020 to May 2021, and blood samples were collected from individuals at high risk of acquiring tick-borne diseases from March 2022 to August 2023. The pooled DNA samples of ticks were analyzed to reveal the presence of tick-borne bacteria using high-throughput sequencing of the 16S rDNA V3–V4 region, and species-specific polymerase chain reaction (PCR) related to sequencing was performed to confirm the presence of pathogenic bacteria in individual ticks and human blood samples. All sera samples were examined for anti-SFGR using ELISA and anti-B. burgdorferi using IFA and WB. A total of 295 ticks (282 Ixodes persulcatus and 13 Dermacentor silvarum) and 245 human blood samples were collected. Rickettsia, Anaplasma, Borrelia miyamotoi, and Coxiella endosymbiont were identified in I. persulcatus by high-throughput sequencing, while Candidatus R. tarasevichiae (89.00%, 89/100), B. garinii (17.00%, 17/100), B. afzelii (7.00%, 7/100), and B. miyamotoi (7.00%, 7/100) were detected in I. persulcatus, as well the dual co-infection with Candidatus R. tarasevichiae and B. garinii were detected in 13.00% (13/100) of I. persulcatus. Of the 245 individuals, B. garinii (4.90%, 12/245), R. slovaca (0.82%, 2/245), and C. burnetii (0.41%, 1/245) were detected by PCR, and the sequences of the target genes of B. garinii detected in humans were identical to those detected in I. persulcatus. The seroprevalence of anti-SFGR and anti-B. burgdorferi was 5.71% and 13.47%, respectively. This study demonstrated that Candidatus R. tarasevichiae and B. garinii were the dominant tick-borne bacteria in I. persulcatus from Arxan, and that dual co-infection with Candidatus R. tarasevichiae and B. garinii was frequent. This is the first time that B. miyamotoi has been identified in ticks from Arxan and R. solvaca has been detected in humans from Inner Mongolia. More importantly, this study demonstrated the transmission of B. garinii from ticks to humans in Arxan, suggesting that long-term monitoring of tick-borne pathogens in ticks and humans is important for the prevention and control of tick-borne diseases.

Extensive spread of tick-borne diseases poses a significant problem for public health and the health of the population living in endemic areas.The aim of the study was to search, analyze genetic material and identify new viral agents of the Phenuiviridae family in taiga ticks collected in Asian regions of Russia using the method of high throughput sequencing.Materials and methods. The study involved 1460 taiga ticks collected in suburban areas of the Tomsk, Yekaterinburg and Primorsky Territory. The genetic material isolated from ticks was sequenced using Illumina technology followed by phylogenetic analysis.Results and discussion. Analysis of the sequencing results made it possible to detect extended nucleotide sequences of the L-gene fragment characteristic of the Phenuiviridae family viruses. We were able to identify 20 nucleotide sequences the length of 250 bp on average in homogenates of Ixodes persulcatus ticks. Eighteen isolates have been identified as members of the genus Uukuvirus and two isolates have been assigned to the genus Phlebovirus, Phenuiviridae family. Phylogenetic analysis has shown that all isolates of the genus Uukuvirus fall under the cluster of Tacheng tick virus 2 belonging to the species Tacheng uukuvirus. They form a separate phylogenetic group which is closely related to two Romanian variants of 2019. Tacheng tick virus 2 was detected in all three surveyed regions of the Asian part of Russia. Two Tomsk isolates of phlebovirus were classified as Sara tick phlebovirus and they clustered with two isolates of phleboviruses from Karelia. Thus, the genetic material of Tacheng tick virus 2 and Sara tick phlebovirus belonging to two genera of the family Phenuiviridae was found in I. persulcatus ticks collected in three geographically different regions of the Asian part of Russia.

Background: California hare coltivirus (CHCV) was isolated in California in 1976 from a hare. Despite its long history, it remained unclear whether CHCV was exclusively distributed in California with limited host ranges. Main body: By next-generation sequencing (NGS), we obtained a complete sequence of CHCV from Ixodes persulcatus collected in 2019 in northeast China. An expanded epidemiological investigation was subsequently performed on ticks belonging to four species (Ix. persulcatus, Haemaphysalis concinna, Devmacentor silvarum, Haemaphysalis longicornis) collected in northeastern China by applying CHCV-specific RT-PCR and sequencing. CHCV RNA-positive results were found in 1.56% of the tick samples. Positive ticks were obtained in three of four sampled locations, with the highest rate observed in Inner Mongolia (2.69%), followed by Heilongjiang (1.94%) and Jilin provinces (0.55%). All positive results were derived from Ix. persulcatus ticks (2.33%), while no positive detection was found in the other tick species, even at the same location. Sequence analysis revealed that the current CHCV showed a high genetic identity (>80% amino acid identity) with the previously reported CHCV in all segments except segment seven (64.59% amino acid identity). Phylogenetic analysis based on RNA-dependent RNA polymerase (RdRp) amino acid sequences demonstrated that both the current and previously reported CHCV strains were grouped phylogenetically into the genus Coltivirus. Both CHCV strains formed a distinct clade, clustering with three human pathogenic coltiviruses (Colorado tick fever virus, Salmon River virus, and Eyach virus), and were distant from the other coltiviruses. Conclusions: We report the identification and characterization of CHCV for the first time in Ix. persulcatus ticks, expanding the currently known geographic scope, host, and genetic heterogeneity in CHCV.

Environmental and climatic changes in northern Europe have shaped a geographical area in which new tick species may become established and introduce new tick-borne pathogens. In recent decades, ticks have expanded their latitudinal and altitudinal range limits in northern Sweden. In this study, ticks were collected in 2018 and 2019 in northern Sweden from different hosts, mainly from dogs, cats and humans. The ticks in 2018 (n = 2141, collected from 65 municipalities in 11 provinces) were identified as Ixodes ricinus (n = 2108, 98.5%), Ixodes persulcatus (n = 18, 0.8%), Ixodes trianguliceps (n = 14, 0.7%) and Hyalomma marginatum (n = 1, 0.05%). The ticks collected in 2019 (n = 519, across a smaller area than in 2018, i.e. Sweden's four northernmost provinces) were identified as I. ricinus (n = 242, 46.6%) and I. persulcatus (n = 277, 53.4%). Among those collected in 2019, the majority of I. ricinus (n = 111, 45.9%) were submitted from the province of Västerbotten, while most I. persulcatus (n = 259, 93.5%) were collected in the province of Norrbotten. This study provides updated figures on the geographical distribution of two Ixodes species in northern Sweden. The results confirmed I. ricinus to be the dominant species and that I. persulcatus has enlarged its distributional area compared with previous reports. Updated knowledge of tick distribution is fundamental for the creation of risk maps and will allow relevant advice to be provided to the general public, suggesting measures to prevent tick bites and consequently tick-borne diseases.

We identified Yezo virus infection in a febrile patient who had a tick bite in northeastern China, where 0.5% of Ixodes persulcatus ticks were positive for viral RNA. Clinicians should be aware of this potential health threat and include this emerging virus in the differential diagnosis for tick-bitten patients in this region.

Abstract 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.

The increasing incidence and range expansion of tick-borne diseases have caused global threats to human and animal health under the background of climate and socioeconomic changes. As an efficient vector in transmission of tick-borne diseases, a growing burden caused by Ixodes persulcatus and associated pathogens could not be underestimated. This study summarized the distribution, hosts, and pathogens of I. persulcatus, and predicted the suitable habitats of this tick species worldwide. An integrated database involving a field survey, reference book, literature review, and related website was constructed. Location records of I. persulcatus and associated pathogens were incorporated into distribution maps using ArcGIS software. Positive rates for I. persulcatus-associated agents were estimated by meta-analysis. The global distribution of the tick species was predicted using Maxent model. I. persulcatus was distributed in 14 countries across the Eurasian continent, involving Russia, China, Japan, and several Baltic Sea states, which ranged between 21°N to 66°N. The tick species fed on 46 species of hosts, and 51 tick-borne agents could be harbored by I. persulcatus. The predictive model showed that I. persulcatus could be predominantly distributed in northern Europe, western Russia, and northern China. Our study fully clarified the potential public health risks posed by I. persulcatus and I. persulcatus-borne pathogens. Surveillance and control measures of tick-borne diseases should be enhanced to promote the health of humans, animals, and ecosystems.

The taiga tick Ixodes persulcatus (Schulze, 1930) (Acari, Ixodidae) is the main vector of the tick-borne encephalitis virus and one of the most widespread species of ixodid ticks in the Palaearctic. In this paper, we present long-term data on the seasonal activity of adult ticks in the north-west of their distribution. The seasonal activity of Ixodes persulcatus was studied from 1982 to 1990 and from 2012 to 2023 in the middle taiga subzone of Karelia (N62.0697, E33.961). In the study area, adult ticks I. persulcatus demonstrate a pronounced spring–summer activity with a unimodal curve of abundance change. A comparison of the monitoring data from the 1980s and the 2010s showed a significant increase in the abundance of I. persulcatus in the study area. A tendency towards an earlier start of the tick activity, as compared to the 1980s, is now being observed.

In recent years, numerous viruses have been identified from ticks, and some have been linked to clinical cases of emerging tick-borne diseases. Chinese northeast frontier is tick infested. However, there is a notable lack of systematic monitoring efforts to assess the viral composition in the area, leaving the ecological landscape of viruses carried by ticks not clear enough. Between April and June 2017, 7101 ticks were collected to perform virus surveillance on the China–North Korea border, specifically in Tonghua, Baishan, and Yanbian. A total of 2127 Ixodes persulcatus were identified. Further investigation revealed the diversity of tick-borne viruses by transcriptome sequencing of Ixodes persulcatus. All ticks tested negative for tick-borne encephalitis virus. Transcriptome sequencing expanded 121 genomic sequence data of 12 different virus species from Ixodes persulcatus. Notably, a new segmented flavivirus, named Baishan Forest Tick Virus, were identified, closely related to Alongshan virus and Harz mountain virus. Therefore, this new virus may pose a potential threat to humans. Furthermore, the study revealed the existence of seven emerging tick-borne viruses dating back to 2017. These previously identified viruses included Mudanjiang phlebovirus, Onega tick phlebovirus, Sara tick phlebovirus, Yichun mivirus, and three unnamed viruses (one belonging to the Peribunyaviridae family and the other two belonging to the Phenuiviridae family). The existence of these emerging tick-borne viruses in tick samples collected in 2017 suggests that their history may extend further than previously recognized. This study provides invaluable insights into the virome of Ixodes persulcatus in the China–North Korea border region, enhancing our ongoing efforts to manage the risks associated with tick-borne viruses.

Mudanjiang phlebovirus (MJPV) is a newly discovered phlebovirus, initially detected from Ixodes persulcatus ticks in China in 2022. In this study, by next-generation sequencing (NGS) on a wide variety of ticks and wild small animals in China, we detected MJPV from I. persulcatus and Meriones meridianus. Additionally, we conducted RT-PCR and sequencing on 1815 adult ticks and 805 wild small mammals collected from eight provinces in China between 2017 and 2021. MJPV RNA-positive results were found in 0.22% (4/1815) of tick samples, as well as in 0.12% (1/805) of rodent samples. All positive detections were obtained from Heilongjiang and Inner Mongolia. Sequencing analysis revealed nucleotide similarities ranging from 98.23% to 99.11%, as well as amino acid similarities ranging from 99.12% to100%, between the current MJPV strain and previously reported strains of MJPV. Phylogenetic tree analysis demonstrated that the previously reported MJPV strain along with our two variants clustered together with other tick-borne phenuiviruses, indicating their close relationship within this viral group. This study represents the first detection of MJPV infection in wild rodents, expanding the known host range for this virus in the endemic regions.

Lyme-borreliosis is a zoonotic, infectious disease that has a complex chain of transmission of the pathogen Borrelia burgdorferi sensu lato and includes the relationship between ixodid ticks, vertebrate hosts, humans and companion animals in the environment. The article shows general trends in the prevalence of canine Lyme-borreliosis in Ukraine depending on environmental, climatic and physiographic factors. The results of a comparative cartographic analysis of the prevalence of Lyme borreliosis among domestic dogs in Ukraine are presented by systematizing, mathematical and statistical processing of the data obtained by surveying veterinarians engaged in clinical veterinary practice. The paper includes generalized data on the clinical manifestations, methods of diagnosis and treatment of Lyme borreliosis in dogs. We determined the dependence of the prevalence of Lyme borreliosis in dogs on the types of physical and geographical territories – natural zones of Ukraine. Each of the natural zones differs in types of relief, climatic conditions, soil types, composition of fauna and flora, which affect the epizootic chain of Lyme disease. There is a clear correlation between the incidence of Lyme borreliosis in dogs and the types of natural areas. The highest incidence of Lyme borreliosis in dogs was observed in the forest-steppe zone and the zone of broad-leaved forests. A sharp decrease in the incidence of dogs was recorded in areas of mixed forests, the Ukrainian Carpathians and in the South of Ukraine in the steppe zone. The study also confirmed that the prevalence of Lyme disease among domestic dogs was influenced by the climatic factors, in particular: gross moisture of territories, average annual air temperature and soil temperature. In Ukraine, veterinarians in the vast majority of cases use serological diagnostic methods: immunochromatographic analysis, immunoenzymatic assay, and western blot, which are insufficient, since the presence of antibodies to the Lyme borreliosis pathogen is only a confirmation of the animal's contact with the antigen and may not indicate the presence of the disease in the clinical form. The generalized data on the use of antimicrobial drugs in the treatment of Lyme borreliosis in dogs indicate the predominant use of tetracycline antibiotics and cephalosporins. The majority of veterinarians reported symptoms of Lyme arthritis, somewhat fewer reported Lyme nephritis, neuroborreliosis, Lyme carditis and in rare cases, veterinarians observed erythema at the site of tick bite. About half of the veterinarians in Ukraine observed an increase in the incidence of Lyme disease in dogs, indicating a probable deterioration of the epizootic and epidemiological situation regarding Lyme borreliosis, especially in areas with favourable conditions for the circulation of Lyme borreliosis pathogens in natural and urban ecotopes. The results substantiate the need for the further study of the circulation of Borrelia burgdorferi sensu lato and their ability to cause disease in humans and animals, as well as the need to implement the principles of the One Health concept for the control and management of Lyme borreliosis.

Widely distributed Ixodes ricinus and Ixodes persulcatus ticks transmit many pathogens of both medical and veterinary significance. The ranges of these tick species overlap and form large sympatric areas in the East European Plain and Baltic countries. It has previously been shown that crossing I. ricinus and I. persulcatus is possible, resulting in the appearance of sterile hybrids. In the present study, we analyzed the features of this hybrid’s life cycle under laboratory conditions. For this purpose, virgin females of I. ricinus and I. persulcatus ticks were obtained in the laboratory, and hybrid generations of ticks were bred from the reciprocal crossings of these two tick species. According to our data, mating the females of I. ricinus and I. persulcatus with the males of another species leads to a decrease in the engorgement success of the females, a decrease in the number of hatched larvae, and the appearance of a hybrid generation in which both females and males are sterile. Under laboratory conditions at a constant room temperature and under natural daylight, the morphogenetic diapause of the engorged I. persulcatus larvae began in September. For I. persulcatus nymphs, it occurred earlier than for I. ricinus, in October and November, respectively. The hybrids generally repeated the features of the life cycle of the mother species.

Four genospecies from the Borrelia burgdorferi sensu lato complex were detected in Ixodes persulcatus and Ixodes pavlovskyi ticks from Siberia and genetically characterized. The presence of Borrelia spp. in Ixodes apronophorus and Ixodes trianguliceps ticks found in Asia has never been studied. In this study, genetic diversity of B. burgdorferi s.l. was investigated in three I. persulcatus / I. trianguliceps / I. apronophorus sympatric habitats in Western Siberia. Three groups of samples were examined: (i) ticks that were taken from rodents and molted in a laboratory; (ii) non-molted ticks collected from rodents; (iii) specimens from small mammals. Expectedly, Borrelia afzelii and Borrelia bavariensis were detected in I. persulcatus and in small mammals from the studied locations. Borrelia bavariensis was first found in molted I. apronophorus and I. trianguliceps. Identical genovariants of B. bavariensis were found in I. apronophorus, I. trianguliceps, and I. persulcatus. In addition, a new Borrelia genovariant was discovered in non-molted and molted I. apronophorus and non-molted I. persulcatus and I. trianguliceps, as well as in small mammals. This new genovariant was genetically characterized using MLST and single locus sequence analysis, which indicated that the new Borrelia genovariant significantly differs from all known Borrelia species. We propose the name ''Candidatus Borrelia sibirica'' for this putative new species.

Ixodes rici nus and Ixodes persulcatus ticks are the main vectors of tick-borne encephalitis virus (TBEV), which has three main subtypes connected with certain tick species: the European subtype, associated with I. ricinus, and the Siberian and Far-Eastern subtypes, associated with I. persulcatus. Distribution ranges of these species overlap and form large sympatric areas in the East European Plain and Baltic countries. It has previously been shown that crossing of I. ricinus and I. persulcatus is possible, with the appearance of sterile hybrids. Hybridization of ticks can affect not only the spread of ticks but also the properties of natural foci of arbovirus infections, in particular TBEV. In the present study, we analyzed the effectiveness of virus transmission from infected mice to larvae and nymphs and trans-stadial transmission (from larvae to nymph and adult) in I. ricinus, I. persulcatus, and hybrids. For this purpose, we bred a hybrid generation from the crossing of I. persulcatus females and I. ricinus males, and we used the Siberian and European subtypes of TBEV. We showed that after feeding on infected mice, virus prevalence in engorged ticks decreased over time, and after molting, the opposite was true. In hybrids we observed the highest acquisition effectiveness and RNA copy numbers during Siberian TBEV subtype transmission. The efficiency of trans-stadial transmission of both TBEV subtypes was similar in hybrids and parental species. After the second trans-stadial TBEV transmission, a significant increase in ticks’ infection rates was observed only in specific subtype-tick combination. Our data demonstrate the possible features of TBEV circulation in the I. ricinus and I. persulcatus sympatry area.

The purpose of the research is to identify ixodid ticks and their pathogens in the left bank area of Central Yakutia. Materials and methods. The work was carried out in 2019–2020 in the left bank area of Central Yakutia. Nine ticks were collected in 2019, and 27 ticks in 2020. We studied forest shrub stations, steppe stations, meadow field stations, near-water stations and stations of settlements. To determine faunal and ecological characteristics of ectoparasites in the territory, we used standard collection methods. The tick species was determined using morphological keys by N. A. Filippova; the determination correctness was confirmed by the PCR method. The collected ticks were studied for causative agents of babesiosis and tick-borne viral encephalitis using PCR analysis. Results and discussion. One species of ixodid ticks, Ixodes persulcatus, inhabits the left bank area of Central Yakutia. Haemaphysalis concinna was not found in Yakutia. In 2008, a natural focus of blood protozoan disease of domestic reindeer appeared for the first time in Yakutia in its central zone. Recently, an increase in the number of I. persulcatus has been observed. Tick activity is recorded from the second decade of May to the second decade of August. The ground-squirrel Spermophilus parryii is the main host for the preimaginal stages. Pathogens were not detected when ticks were examined for causative agents of babesiosis and tick-borne viral encephalitis using PCR analysisd.

The emerging Babesia venatorum upsurges as a potential health threat occurring in China and other endemic countries. Few attempts to isolate and culture the Babesia species had been conducted in vitro. We collected the questing Ixodes persulcatus from identified endemic areas and allowed them to feed on the severe combined immunodeficiency (SCID) mice. The positive mice were chosen to provide positive erythrocytes with asexual B. venatorum for continuous culture in mouse or human erythrocytes in vitro, with RPMI 1640 medium and appropriate serum. With B. venatorum in the SCID mice, erythrocytes were cultured in vitro for confirmation by morphological observations with transmission electron microscopes. Sequences of B. venatorum were then identified by way of conventional PCR amplification. Parasitemia counts monitored the growth of B. venatorum on thin blood smears and real-time quantitative PCR in parallel. As expected, B. venatorum positive mice were harvested by sufficient attacks of I. persulcatus ticks. The erythrocytes of the infected mice were then inoculated and successfully cultured in donated erythrocytes from humans and mice in RPMI 1640 culture medium. Active growth of B. venatorum was well demonstrated in human erythrocytes with 3.3 times parasite-load when compared with a mouse under similar conditions. With the increased subcultures, a prolonged period of detectable parasitemia with much higher peak parasitemia and shorter time to reach peak parasitemia were observed in the following subcultures. A new strategy for isolation and in vitro culture of B. venatorum has been provided with a continuous supply of sufficient pathogens to satisfy human babesiosis's testings and clinical therapies.

(1) Background: Amplicon-based 16S rRNA profiling is widely used to study whole communities of prokaryotes in many niches. Here, we comparatively examined the microbial composition of three tick species, Ixodes ricinus, Ixodes persulcatus and Dermacentor reticulatus, which were field-collected in Latvia. (2) Methods: Tick DNA samples were used for microbiome analysis targeting bacterial 16S rDNA using next-generation sequencing (NGS). (3) Results: The results showed significant differences in microbial species diversity and composition by tick species and life stage. A close similarity between microbiomes of I. ricinus and I. persulcatus ticks was observed, while the D. reticulatus microbiome composition appeared to be more distinct. Significant differences in alpha and beta microbial diversity were observed between Ixodes tick life stages and sexes, with lower taxa richness indexes obtained for female ticks. The Francisella genus was closely associated with D. reticulatus ticks, while endosymbionts Candidatus Midichlorii and Candidatus Lariskella were associated with I. ricinus and I. persulcatus females, respectively. In I. ricinus females, the endosymbiont load negatively correlated with the presence of the Rickettsia genus. (4) Conclusions: The results of this study revealed important associations between ticks and their microbial community and highlighted the microbiome features of three tick species in Latvia.

Abstract Background Borrelia miyamotoi is a newly described relapsing fever spirochete transmitted by ixodid tick species. Little is known about the prevalence of B. miyamotoi infections in humans and ticks in Inner Mongolia, China. Therefore, we investigated the prevalence of B. miyamotoi in Ixodes persulcatus ticks, and we aimed to isolateB. miyamotoi from I. persulcatus from four regions of Greater Khingan, Inner Mongolia, China. Methods From May to June each year during the period 2016–2019, host-seeking adult I. persulcatus ticks were collected from vegetation. Genomic DNA was prepared from half of each tick body for PCR template, and the remaining half was used to cultivate B. miyamotoi in BSK-M medium. We employed quantitative real-time PCR (qPCR) to detect Borrelia DNA in the ticks and to calculate the prevalence of B. miyamotoi and infections with other borreliae. For characterization of the isolated B. miyamotoi, we performed draft genome sequencing and multilocus sequencing analysis (MLSA). Results A total of 2656 adult I. persulcatus ticks were collected. The overall prevalence of relapsing fever (RF) borreliae in ticks was 5.0% (134/2656) and that of Lyme disease (LD) borreliae was 43.8% (1164/2656). Co-infection with RF and LD borreliae was observed in 63 ticks (2.4%). Ticks that were positive for RF borreliae by qPCR were subjected to glycerophosphodiester diester phosphodiesterase gene (glpQ) PCR amplification and sequencing, through which we identified the RF borrelia specimens as B. miyamotoi. Furthermore, the B. miyamotoi strain Hetao-1 was isolated from I. persulcatus, and a draft genome sequence was obtained from the isolate. Sequencing determined the strain Hetao-1 genome to be approximately 906.1 kbp in length (28.9% average GC content), and MLSA identified the strain as ST633, which has previously been reported in Japan and Mongolia. Conclusion We detected B. miyamotoi from I. persulcatus ticks collected in Inner Mongolia, and successfully isolated a B. miyamotoi strain. To our knowledge, this is the first study to culture a B. miyamotoi isolate from China. The data on the prevalence of B. miyamotoi and other borreliae in I. persulcatus ticks will be fundamental for future epidemiological studies of B. miyamotoi disease in Inner Mongolia.

Abstract Background Hard ticks act as arthropod vectors in the transmission of human and animal pathogens and are widely distributed in northern China. The aim of this study is to screen the important tick-borne pathogens (TBPs) carried by hard ticks in Inner Mongolia using metagenomic next-generation sequencing (mNGS) and to estimate the risk of human infection imposed by tick bites. Methods The adult Dermacentor nuttalli (n = 203) and Ixodes persulcatus (n = 36) ticks feeding on cattle were collected. The pooled DNA samples prepared from these ticks were sequenced as the templates for mNGS to survey the presence of TBPs at the genus level. Individual tick DNA samples were detected by genus--specific or group-specific nested polymerase chain reaction (PCR) of these TBPs and combined with DNA sequencing assay to confirm the results of mNGS. Results R. raoultii (45.32%, 92/203), Candidatus R. tarasevichiae (5.42%, 11/203), Anaplasma sp. Mongolia (26.60%, 54/203), Coxiella-like endosymbiont (CLE) (53.69%, 109/203), and Babesia venatorum (7.88%, 16/203) were detected in D. nuttalli, while R. raoultii (30.56%, 11/36), Anaplasma sp. Mongolia (27.80%, 10/36), and CLE (27.80%, 10/36) were detected in I. persulcatus. The double- and triple-pathogen/endosymbiont co-infections were detected in 40.39% of D. nuttalli and 13.89% of I. persulcatus, respectively. The dual co-infection with R. raoultii and CLE (14.29%, 29/203) and triple co-infection with R. raoultii, Anaplasma sp. Mongolia, and CLE (13.79%, 28/203) were most frequent in D. nuttalli. Conclusions This study provides insight into the microbial diversity of D. nuttalli and I. persulcatus in Inner Mongolia, China, reporting for the first time that Candidatus R. tarasevichiae had been found in D. nuttalli in China, and for the first time in the world that Anaplasma sp. Mongolia has been detected in I. persulcatus. This study proves that various vertically transmitted pathogens co-inhabit D. nuttalli and I. persulcatus, and indicates that cattle in Inner Mongolia are exposed to several TBPs. Graphic Abstract

Mukawa virus (MKWV), a novel tick-borne virus (TBV) of the genus Phlebovirus of family Phenuiviridae, has been firstly reported in Ixodes persulcatus in Japan. In this study, we made an epidemiological investigation in China to obtain the geographic distribution and genetic features of this virus outside Japan. We screened 1,815 adult ticks (665 I. persulcatus, 336 Dermacentor silvarum, 599 Haemaphysalis longicornis, 170 Rhipicephalus microplus, 45 Haemaphysalis concinna) and 805 wild small mammals collected from eight provinces. The positive rate of 6.77% (45/665, including 18 female and 27 male I. persulcatus) and 2.22% (1/45, 1 male H. concinna) were obtained from I. persulcatus and H. concinna in Heilongjiang province, respectively. No evidence of MKWV infection was found in other three tick species or any of the mammalian species. The virus can infect the Vero cells successfully, indicating the ability of MKWV to replicate in mammalian cells. A phylogenetic tree based on the nucleotide sequences of L, M, and S segments demonstrated that the Japanese MKWV variant, our two MKWV variants, and KURV were clustered with the members of the mosquito/sandfly-borne phleboviruses and distant from other tick-borne phenuiviruses. A phylogenetic analysis based on 895 bp partial L gene sequences (n = 46) showed that all MKWV sequences were separated into three lineages. Our results showed the presence of MKWV in I. persulcatus and H. concinna in northeast of China, highlighting the necessity of epidemiological study in wider regions. Due to the ability of MKWV to replicate in mammalian cells, the potential for zoonosis, and wide distribution of I. persulcatus and H. concinna in China, the important vectors of MKWV, further screening to more tick species, wild animals, domestic animals, and humans raises up practical significance.

Abstract Background In the South of Kazakhstan, Almaty Oblastʼ (region) is endemic for tick-borne encephalitis, with 0.16–0.32 cases/100,000 population between 2016–2018. The purpose of this study was to determine the prevalence and circulating subtypes of tick-borne encephalitis virus (TBEV) in Almaty Oblastʼ and Kyzylorda Oblastʼ. Methods In 2015 we investigated 2341 ticks from 7 sampling sites for the presence of TBEV. Ticks were pooled in 501 pools and isolated RNA was tested for the presence of TBEV by RT-qPCR. For the positive samples, the E gene was amplified, sequenced and a phylogenetic analysis was carried out. Results A total of 48 pools were TBEV-positive by the RT-qPCR. TBEV-positive ticks were only detected in three districts of Almaty Oblastʼ and not in Kyzylorda Oblastʼ. The positive TBEV pools were found within Ixodes persulcatus, Haemaphysalis punctata and Dermacentor marginatus. These tick species prevailed only in Almaty Oblastʼ whereas in Kyzylorda Oblastʼ Hyalomma asiaticum and D. marginatus are endemic. The minimum infection rates (MIR) in the sampling sites were 4.4% in Talgar, 2.8% in Tekeli and 1.1% in Yenbekshikazakh, respectively. The phylogenetic analysis of the generated sequences indicates that TBEV strains found in Almaty Oblastʼ clusters in the Siberian subtype within two different clades. Conclusions We provided new data about the TBEV MIR in ticks in Almaty Oblastʼ and showed that TBEV clusters in the Siberian Subtype in two different clusters at the nucleotide level. These results indicate that there are different influences on the circulating TBEV strains in south-eastern Kazakhstan. These influences might be caused by different routes of the virus spread in ticks which might bring different genetic TBEV lineages to Kazakhstan. The new data about the virus distribution and vectors provided here will contribute to an improvement of monitoring of tick-borne infections and timely anti-epidemic measures in Kazakhstan.

Abstract Background Because ixodid ticks are vectors of zoonotic pathogens, including Borrelia, information of their abundance, seasonal variation in questing behaviour and pathogen prevalence is important for human health. As ticks are invading new areas northwards, information from these new areas are needed. Taiga tick (Ixodes persulcatus) populations have been recently found at Bothnian Bay, Finland. We assessed seasonal variation in questing abundance of ticks and their pathogen prevalence in coastal deciduous forests near the city of Oulu (latitudes 64–65°) in 2019. Methods We sampled ticks from May until September by cloth dragging 100 meters once a month at eight study sites. We calculated a density index (individuals/100 m2) to assess seasonal variation. Samples were screened for Borrelia burgdorferi (sensu lato) (including B. afzelii, B. garinii, B. burgdorferi (sensu stricto) and B. valaisana), Borrelia miyamotoi, Anaplasma phagocytophilum, Rickettsia spp., Neoehrlichia mikurensis, Francisella tularensis and Bartonella spp., Babesia spp. and for the tick-borne encephalitis virus. Results All except one nymph were identified as I. persulcatus. The number of questing adults showed a strong peak in May (median: 6.5 adults/100 m2), which is among the highest values reported in northern Europe, and potentially indicates a large population size. After May, the number of questing adults declined steadily with few adults still sampled in August. Nymphs were present from May until September. We found a striking prevalence of Borrelia spp. in adults (62%) and nymphs (40%), with B. garinii (51%) and B. afzelii (63%) being the most common species. In addition, we found that 26% of infected adults were coinfected with at least two Borrelia genospecies, mainly B. garinii and B. afzelii, which are associated with different host species. Conclusions The coastal forest environments at Bothnian Bay seem to provide favourable environments for I. persulcatus and the spread of Borrelia. High tick abundance, a low diversity of the host community and similar host use among larvae and nymphs likely explain the high Borrelia prevalence and coinfection rate. Research on the infestation of the hosts that quantifies the temporal dynamics of immature life stages would reveal important aspects of pathogen circulation in these tick populations.

Abstract Background Tick-borne diseases are of substantial concern worldwide in both humans and animals. Several hard tick species are of medical and veterinary interest in Europe, and changes in the range of tick species can affect the spread of zoonotic pathogens. The aim of the present study was to map the current prevalence and distribution pattern of ticks and related tick-borne pathogens in Latvia, a Baltic state in northern Europe. Methods Nearly 4600 Ixodes ricinus, I. persulcatus and Dermacentor reticulatus tick samples were collected in all regions of Latvia during 2017–2019 and were screened by molecular methods to reveal the prevalence and distribution pattern of a wide spectrum of tick-borne pathogens. Results New localities of D. reticulatus occurrence were found in western and central Latvia, including the Riga region, indicating that the northern border of D. reticulatus in Europe has moved farther to the north. Among the analyzed ticks, 33.42% carried at least one tick-borne pathogen, and 5.55% of tick samples were positive for two or three pathogens. A higher overall prevalence of tick-borne pathogens was observed in I. ricinus (34.92%) and I. persulcatus (31.65%) than in D. reticulatus (24.2%). The molecular analysis revealed the presence of tick-borne encephalitis virus, Babesia spp., Borrelia spp., Anaplasma phagocytophilum and Rickettsia spp. Overall, 15 and 7 tick-borne pathogen species were detected in Ixodes spp. and D. reticulatus ticks, respectively. This is the first report of Borrelia miyamotoi in Latvian field-collected ticks. Conclusions This large-scale countrywide study provides a snapshot of the current distribution patterns of Ixodes and Dermacentor ticks in Latvia and gives us a reliable overview of tick-borne pathogens in Latvian field-collected ticks.

Over the last years, an increasing rate of ixodes tick bites has been registered in the northern regions of the European Russia. In addition, the number of subjects request medical assistance due to tick bites has been dramatically increased in the Komi Republic. In addition, incidence of tick-borne encephalitis was also increased particularly starting since 2009. However, highly limited data on pathogen genetic diversity related to viral tick-borne infections in this region are currently available. Taiga ticks (Ixodespersulcatus) collected from the Komi Republic southern and central part vegetation were examined to identify and genotype tick-borne viruses. Individual ticks were used to identify by RT-PCR viral RNA coupled to tick-borne encephalitis and Kemerovo viruses. Viral genome fragment sequencing allowed to unambiguously identify these viruses. It was found that viral RNA tick-borne encephalitis was detected in 6.8±1.2% individual ticks. Moreover, tick-linked isolate genotyping based on analyzing E protein gene fragment nucleotide sequence derived from tick-borne encephalitis discovered that 35% and 65% isolates belonged to the Far Eastern and Siberian subtype, respectively. In addition, subsequent phylogenetic analysis demonstrated that at least four variants of the Siberian and Far Eastern subtypes of tick-borne encephalitis virus were detected, which were close to the viruses circulating in the Urals and Siberia. In contrast, prevalence of Kemerovo virus in taiga ticks was 0.8±0.2%. Sequencing of Kemerovo virus RNA-dependent RNA polymerase gene fragment showed around 94% homology with the remainder of the Kemerovo virus strains. Phylogenetic analysis of the Kemerovo virus genome fragments demonstrated at least two subtypes circulating in the Komi Republic. Thus, it was suggested that tick-borne encephalitis virus was introduced relatively recently from the Urals and Siberian region into the natural foci of the Komi Republic. Moreover, genetic differences found in Kemerovo virus strains presume for them a longer lasting evolution throughout the natural foci of this region. In addition, a potential role for birds and their ticks in rapid spreading of viral tick-borne infections in the Komi Republic is also discussed. Thus, the data on genetic diversity of the viral agents related to tick-born encephalitis and Kemerovo fever may be useful for improving their diagnostics, prevention and treatment in the Komi Republic.

The sharp rising incidence of tick-borne encephalitis (TBE) in Komi Republic at the North-east of European Russia was recorded last decades. Tick-bite incidence also was grown. Rapid rise of TBE incidence growth and Ixodidae ticks depends on a number of factors, and the impact of climate change being one of them. Ixodes persulcatus ticks is considered as a main vector of TBE in Komi. Our objective is to estimate the influence of air temperature change on the tickbite incidence and Ixodes persulcatus population in Komi. Komi Republic is located near the Polar circle where the northern frontier of Ixodes persulcatus ticks situated and we expected the growth of tick’s population. The number of Komi inhabitants seeking medical care after tick bites in 1992–2014 was considered. Gridded monthly air temperature data with grid size 0.5 degree were recalculated to temperature referred to Komi administrative units. The time series of annual number of tick victims from 1992 till 2014 and model air temperature from 1948 till 2016 for all Komi administrative units were compiled. We analyzed the data on tick-bite incidence in Komi administrative units in relation to changes in local annual average air temperature within the study area. The linear dependence of the tick-bite incidence on air temperature was established when of the tick-bite incidence is represented in logarithm form. The tick population depends not only on temperature but humidity, landcover and hosts. Described areas of Komi belong to humid climate, where precipitations exceed evaporation. Most of the Komi territory is covered by taiga with underwood, grass and bush. Hosts of the first and the major levels are represented by birds and rodents. The dependence of tick-bite incidence and temperature looks like “Malthus’s law”, but the development of population depends on temperature not on time. The exponential growth in the nearest future will ceased and the population will proceed to stable phase. Ticks population in Komi Republic is moving to the North and the air temperature determines the dynamics of population.

Ticks can carry multiple pathogens, and Inner Mongolia’s animal husbandry provides excellent environmental conditions for ticks. This study characterized the microbiome of ticks from different geographical locations in Inner Mongolia; 905 Dermacentor nuttalli and 36 Ixodes persulcatus were collected from sheep in three main pasture areas and from bushes within the forested area. Mixed DNA samples were prepared from three specimens from each region and tick species. Microbial diversity was analyzed by 16S rRNA sequencing, and α and β diversity were determined. The predominant bacterial genera were Rickettsia (54.60%), including Rickettsiales bacterium Ac37b (19.33%) and other Rickettsia (35.27%), Arsenophonus (11.21%), Candidatus Lariskella (10.84%), and Acinetobacter (7.17%). Rickettsia bellii was identified in I. persulcatus, while Rickettsiales bacterium Ac37b was found in D. nuttalli from Ordos and Chifeng. Potential Rickettsia and Anaplasma coinfections were observed in the Ordos region. Tick microbial diversity analysis in Inner Mongolia suggests that sheep at the sampling sites were exposed to multiple pathogens.

Objective of research: To study the spread of causative agents of babesiosis in ticks collected on the territory of the Kirov region with the use of molecular-genetic methods. Materials and methods: In this paper, the method of molecular-genetic detection of genetic material of the pathogen in the sample was used (the method of polymerase chain reaction (PCR). Ticks were collected from vegetation cover as well as from people and domestic animals (dogs, cats). Ticks were estimated according to identification tables. Causative agents of babesiosis were determined by the extraction of total nucleic acids from each tick; all ticks were examined using PCR. Total DNA extraction from ticks stored in 70% ethyl alcohol was performed with the use of guanidine thiocyanate. The proportion of ticks infected with Babesia was determined depending on the year, collection area on the territory of the Kirov region, species and sexual belonging of ticks. Results and discussion: It was found that the major tick vectors of Babesia on the territory of the Kirov region are ticks Ixodes persulcatus . In addition, two other tick species Dermacentor reticulatus and I. ricinus were detected in that region. It was shown, that the rate of Babesia infection in ticks Ixodes persulcatus was higher than in D. reticulatus and I. ricinus . The average percentage of ticks infected with Babesia was 53,07 %. A gradual increase of percentage of infected individuals with the maximum value 73,2 % was registered in 2012. However, in the following year, a significant decrease (51,7 %) was reported, and the minimum number of ticks infected with Babesia was observed in 2014 (37,4 %). A significant increase in infestation of ticks was newly observed in 2015 (50,4 %). It was found that male and female ticks are equally likely to be vectors for Babesia. Infection rates in male and female ticks were 54,5 and 49,3 %, respectively. Fluctuations in the number of infected ticks depending on the year (maximum in 2012 and minimum in 2014) were determined. Ticks from South-West districts of the region are mostly infected with Babesia. In the city of Kirov, 55,8 % of ticks are infected with Babesia in spite of acaricide treatment.

Introduction. The detection of the first cases of tick-borne human granulocytic anaplasmosis in Russia, discovery of genetic markers for Anaplasma spp. in ixodid ticks and reporting of a significant number of cases of tick-borne infections in the southern part of Western Siberia give reason to suppose that causative agents of tick-borne anaplasmosis may be transmitted in Tomsk and its suburbs. Objective. To study the distribution and species biodiversity of A. phagocytophilum in ixodid ticks in Tomsk Region. Materials and methods. The analysis of 690 individual ixodid ticks (larvae and adults) was carried out for Ixodes persulcatus (n = 530) and Dermacentor reticulatus (n = 160) ticks collected in 2015–2016 on the territory of urban and suburban biotopes of Tomsk. Primary screening of ticks for the presence of genetic material of A. phagocytophilum was conducted using two-round PCR with species-specific primers for the 16S rRNA gene. The amplification (1,220 kB) of the groESL fragment of the heat shock protein operon was performed for positive isolates with subsequent determination of the nucleotide sequence in the gene fragment for phylogenetic analysis. Results. The number of A. phagocytophilum positive samples for I. persulcatus (larvae) was 1.2 ± 0.6%, I. persulcatus (adult) was 1.8 ± 0.7%; and D. reticulatus (adult) was 0.6 ± 0.3%. Analysis of the nucleotide sequence of the gene fragments in groESL operon for nine isolates confirmed that the genetic material of the granulocytic anaplasmosis was detected. Phylogenetic analysis showed that all the isolates belonged to the first group of the “new cluster” of A. phagocytophilum. Conclusion. The causative agent of human granulocytic anaplasmosis has been newly detected in I. persulcatus ticks collected in urban and suburban biotopes of Tomsk and in D. reticulatus from urban foci.

Abstract The taiga tick, Ixodes persulcatus, has previously been limited to eastern Europe and northern Asia, but recently its range has expanded to Finland and northern Sweden. The species is of medical importance, as it, along with a string of other pathogens, may carry the Siberian and Far Eastern subtypes of tick-borne encephalitis virus. These subtypes appear to cause more severe disease, with higher fatality rates than the central European subtype. Until recently, the meadow tick, Dermacentor reticulatus, has been absent from Scandinavia, but has now been detected in Denmark, Norway and Sweden. Dermacentor reticulatus carries, along with other pathogens, Babesia canis and Rickettsia raoultii. Babesia canis causes severe and often fatal canine babesiosis, and R. raoultii may cause disease in humans. We collected 600 tick nymphs from each of 50 randomly selected sites in Denmark, southern Norway and south-eastern Sweden in August–September 2016. We tested pools of 10 nymphs in a Fluidigm real time PCR chip to screen for I. persulcatus and D. reticulatus, as well as tick-borne pathogens. Of all the 30,000 nymphs tested, none were I. persulcatus or D. reticulatus. Our results suggest that I. persulcatus is still limited to the northern parts of Sweden, and have not expanded into southern parts of Scandinavia. According to literature reports and supported by our screening results, D. reticulatus may yet only be an occasional guest in Scandinavia without established populations.

Ixodes ricinus and Ixodes persulcatus ticks are the main vectors of tick-borne encephalitis virus and some bacterial pathogens. The regions where these tick species live overlap, forming large sympatric areas. It has previously been shown that these tick species have no morphological barrier, and interspecies crossing is possible with the appearance of sterile hybrids. It has also been shown that hybrid larvae and nymphs can be differentiated using discriminant functions based on a set of morphological features. However, such an approach is laborious and rather ineffective with adult ticks, making a molecular approach necessary. In the current work, we tested the ability of different systems to differentiate laboratory-obtained hybrid ticks. Our data suggest that commonly used primer sets that target rRNA are unsuitable for hybrid tick determination, likely due to the rRNA region being linked with the X chromosome in I. ricinus and I. persulcatus ticks. We tested several primer sets targeting different non rRNA genes to assess their ability to determine hybrids. The best primer set, Toll_R, targeting the putative Toll gene, showed little to no bias when used for DNA amplification from hybrid ticks. Thus, Toll gene can be further used for hybrid detection.

Spiroplasma are vertically-transmitted endosymbionts of ticks and other arthropods. Field-collected Ixodes persulcatus have been reported to harbour Spiroplasma, but nothing is known about their persistence during laboratory colonisation of this tick species. We successfully isolated Spiroplasma from internal organs of 6/10 unfed adult ticks, belonging to the third generation of an I. persulcatus laboratory colony, into tick cell culture. We screened a further 51 adult male and female ticks from the same colony for presence of Spiroplasma by genus-specific PCR amplification of fragments of the 16S rRNA and rpoB genes; 100% of these ticks were infected and the 16S rRNA sequence showed 99.8% similarity to that of a previously-published Spiroplasma isolated from field-collected I. persulcatus. Our study shows that Spiroplasma endosymbionts persist at high prevalence in colonised I. persulcatus through at least three generations, and confirms the usefulness of tick cell lines for isolation and cultivation of this bacterium.

Ticks transmit various pathogens, including parasites, bacteria and viruses to humans and animals. To investigate the ticks and the potentially zoonotic pathogens that they may carry, questing ticks were collected in 2017 from 7 sites in Tokachi District, eastern Hokkaido, Japan. A total of 1563 ticks including adults (male and female), nymphs and larvae were collected. Four species of ticks were identified: Ixodes ovatus, Ixodes persulcatus, Haemaphysalis japonica and Haemaphysalis megaspinosa. Of the 1563 ticks, 1155 were used for DNA extraction. In total, 527 individual tick DNA samples prepared from adults (n = 484), nymphs (n = 41) and larvae (n = 2); and 67 pooled tick DNA samples prepared from larval stages (n = 628) were examined using PCR methods and sequencing to detect Borrelia burgdorferi (sensu lato) and Rickettsia spp. The phylogenetic analysis of Borrelia spp. flaB gene sequences showed the presence of the human pathogenic B. burgdorferi (s.l.) species (Borrelia garinii, Borrelia bavariensis and Borrelia afzelii) in I. persulcatus, whereas the non-pathogenic species Borrelia japonica was found only in I. ovatus. In I. persulcatus, B. garinii and/or its closely related species B. bavariensis was detected in both adults and nymphs at a prevalence of 21.9% whereas B. afzelii was found only in adults (1.8%). The prevalence of B. japonica in adult I. ovatus was 21.8%. Rickettsia species were identified through phylogenetic analysis based on gltA, 16S rRNA, ompB and sca4 genes. Four genotypes were detected in the samples which were classified into three species. The prevalence of human pathogenic Rickettsia helvetica was 26.0% in I. persulcatus adults and nymphs, 55.6% in I. persulcatus larval pools, and 1.7% in H. megaspinosa larval pools. The prevalence of “Candidatus R. tarasevichiae” was 15.4% in I. persulcatus adults and nymphs and 33.3% in I. persulcatus larval pools. The prevalence of “Candidatus R. principis” in H. megaspinosa adults and nymphs was 11.1% whereas it was detected in 3.4% of the H. megaspinosa larval pools. These results indicate that most of the risks of Lyme borreliosis and spotted fever group rickettsiosis infection in eastern Hokkaido, Japan, are restricted to I. persulcatus.

Abstract Background Almost 3500 tick samples, originally collected via a nationwide citizen science campaign in 2015, were screened to reveal the prevalence and distribution of a wide spectrum of established and putative tick-borne pathogens vectored by Ixodes ricinus and I. persulcatus in Finland. The unique geographical distribution of these two tick species in Finland allowed us to compare pathogen occurrence between an I. ricinus-dominated area (southern Finland), an I. persulcatus-dominated area (northern Finland), and a sympatric area (central Finland). Results Of the analysed ticks, almost 30% carried at least one pathogen and 2% carried more than one pathogen. A higher overall prevalence of tick-borne pathogens was observed in I. ricinus than in I. persulcatus: 30.0% (604/2014) versus 24.0% (348/1451), respectively. In addition, I. ricinus were more frequently co-infected than I. persulcatus: 2.4% (49/2014) versus 0.8% (12/1451), respectively. Causative agents of Lyme borreliosis, i.e. bacterial genospecies in Borrelia burgdorferi (sensu lato) group, were the most prevalent pathogens (overall 17%). “Candidatus Rickettsia tarasevichiae” was found for the first time in I. ricinus ticks and in Finnish ticks in general. Moreover, Babesia divergens, B. venatorum and “Candidatus Neoehrlichia mikurensis” were reported for the first time from the Finnish mainland. Conclusions The present study provides valuable information on the prevalence and geographical distribution of various tick-borne pathogens in I. ricinus and I. persulcatus ticks in Finland. Moreover, this comprehensive subset of ticks revealed the presence of rare and potentially dangerous pathogens. The highest prevalence of infected ticks was in the I. ricinus-dominated area in southern Finland, while the prevalence was essentially equal in sympatric and I. persulcatus-dominated areas. However, the highest infection rates for both species were in areas of their dominance, either in south or north Finland.

Abstract Background The Ixodes pavlovskyi tick species, a member of the I. persulcatus/I. ricinus group, was discovered in the middle of the 20th century in the Russian Far East. Limited data have been reported on the detection of infectious agents in this tick species. The aim of this study was to investigate the prevalence and genetic variability of a wide range of infectious agents in I. pavlovskyi ticks collected in their traditional and recently invaded habitats, the Altai Mountains and Novosibirsk Province, respectively, which are both located within the Western Siberian part of the I. pavlovskyi distribution area. Results This study reports the novel discovery of Borrelia bavariensis, Rickettsia helvetica, R. heilongjiangensis, R. raoultii, “Candidatus Rickettsia tarasevichiae”, Anaplasma phagocytophilum, Ehrlichia muris, “Candidatus Neoehrlichia mikurensis” and Babesia microti in I. pavlovskyi ticks. In addition, we confirmed the previous identification of B. afzelii, B. garinii and B. miyamotoi, as well as tick-borne encephalitis and Kemerovo viruses in this tick species. The prevalence and some genetic characteristics of all of the tested agents were compared with those found in I. persulcatus ticks that were collected at the same time in the same locations, where these tick species occur in sympatry. It was shown that the prevalence and genotypes of many of the identified pathogens did not significantly differ between I. pavlovskyi and I. persulcatus ticks. However, I. pavlovskyi ticks were significantly more often infected by B. garinii and less often by B. bavariensis, B. afzelii, “Ca. R. tarasevichiae”, and E. muris than I. persulcatus ticks in both studied regions. Moreover, new genetic variants of B. burgdorferi (sensu lato) and Rickettsia spp. as well as tick-borne encephalitis and Kemerovo viruses were found in both I. pavlovskyi and I. persulcatus ticks. Conclusion Almost all pathogens that were previously detected in I. persulcatus ticks were identified in I. pavlovskyi ticks; however, the distribution of species belonging to the B. burgdorferi (sensu lato) complex, the genus Rickettsia, and the family Anaplasmataceae was different between the two tick species. Several new genetic variants of viral and bacterial agents were identified in I. pavlovskyi and I. persulcatus ticks.

Kemerovo virus (KEMV), a member of Reoviridae family, Orbivirus genus, is transmitted by ixodes ticks and can cause the damage of human central nervous system. The occurrence and genetic diversity of KEMV in Western Siberia still remained poorly studied, so, the aim of this work was to investigate the prevalence and genetic variability of KEMV in ixodid ticks from Western Siberia. A total of 1958 Ixodes persulcatus, I. pavlovskyi ticks and their hybrids from Novosibirsk and Omsk provinces, Altai Republic (Russia) and East Kazakhstan province (Kazakhstan) were analyzed for the presence of KEMV and tick-borne encephalitis virus (TBEV) RNA. It was shown that KEMV distribution area in Western Siberia was wider and included Northern and Northeastern Altai in addition to Omsk and Novosibirsk provinces. For the first time this virus was found in Kazakhstan. The occurrence of KEMV was statistically lower than TBEV in most locations in Western Siberia. KEMV was found both in I. persulcatus and I. pavlovskyi ticks and in their hybrids. Notably, KEMV variants found in 2010s genetically differ from those isolated in 1960s. Moreover, the possibility of reassortment for KEMV was demonstrated for the first time.

A national crowdsourcing-based tick collection campaign was organized in 2015 with the objective of producing novel data on tick distribution and tick-borne pathogens in Finland. Nearly 20 000 Ixodes ticks were collected. The collected material revealed the nationwide distribution of I. persulcatus for the first time and a shift northwards in the distribution of I. ricinus in Finland. A subset of 2038 tick samples containing both species was screened for Borrelia burgdorferi sensu lato (the prevalence was 14.2% for I. ricinus and 19.8% for I. persulcatus), B. miyamotoi (0.2% and 0.4%, respectively) and tick-borne encephalitis virus (TBEV; 0.2% and 3.0%, respectively). We also report new risk areas for TBEV in Finland and, for the first time, the presence of B. miyamotoi in ticks from mainland Finland. Most importantly, our study demonstrates the overwhelming power of citizen science in accomplishing a collection effort that would have been impossible with the scientific community alone.Emerging Microbes & Infections (2017) 6, e31; doi:10.1038/emi.2017.17; published online 10 May 2017

Blood samples were taken from 541 small mammal captured in 2013-2015 in Znamensky district of Omsk region from Ixodes persulcatus and Ixodes trianguliceps sympatric area and examined for the Babesia spp. presence by nested PCR with subsequent sequencing of positive samples. Babesia microti DNA was found in 31,1 % of positive samples; a proportion of infected mammals varied from 5,3 % to 61,6 % in different sampling periods. B. microti DNA was found in samples from three prevailing Myodes species as well as from a root vole (Microtus oeconomus), field voles (Microtus argestis) and Siberian chipmunks (Tamias sibiricus). It was shown that identified B. microti samples belong to two genetic groups: B. microti 'US'-type and B. microti 'Munich'-type; notably that > 90 % infected mammals contained DNA of nonpathogenic for human B. microti 'Munich'-type. We suppose that I. trianguliceps tick is the most probable vector of B. microti 'Munich'-type.

The aim was to study the species composition, abundances and distribution of ixodid ticks in the City of Petrozavodsk. Samples were collected in May–September, 2006–2014 using two methods simultaneously: by flagging from vegetation and by collection from dogs. In addition, 47 tick specimens were provided by veterinary clinics based in Petrozavodsk. Records from the entire study period include three tick species: Dermacentor marginatus, Ixodes persulcatus and Ixodes ricinus. A substantial share in the samples belonged to Ixodes persulcatus – 344 specimens. I. ricinus (5 females) was identified only in the material coming from veterinary clinics. The only female D. marginatus was obtained from a dog from a route through the central part of the city. Samples collected by flagging contained only I. persulcatus, its relative abundance along the control transect (Botanical Garden, suburbs) ranging within 0.9–22 specimens per flag-km. I. persulcatus abundance within city limits was low (

In recent decades, many new flavi-like viruses have been discovered predominantly in different invertebrates and, as was recently shown, some of them may cause disease in humans. The Jingmenvirus (JMV) group holds a special place among flaviviruses and flavi-like viruses because they have a segmented ssRNA(+) genome. We detected Alongshan virus (ALSV), which is a representative of the JMV group, in ten pools of adult Ixodes persulcatus ticks collected in two geographically-separated Russian regions. Three of the ten strains were isolated in the tick cell line IRE/CTVM19. One of the strains persisted in the IRE/CTVM19 cells without cytopathic effect for three years. Most ALSV virions purified from tick cells were spherical with a diameter of approximately 40.5 nm. In addition, we found smaller particles of approximately 13.1 nm in diameter. We obtained full genome sequences of all four segments of two of the isolated ALSV strains, and partial sequences of one segment from the third strain. Phylogenetic analysis on genome segment 2 of the JMV group clustered our novel strains with other ALSV strains. We found evidence for the existence of a novel upstream open reading frame in the glycoprotein-coding segment of ALSV and other members of the JMV group.

We confirmed infection of 2 patients with Borrelia miyamotoi in Japan by retrospective surveillance of Lyme disease patients and detection of B. miyamotoi DNA in serum samples. One patient also showed seroconversion for antibody against recombinant glycerophosphodiester phosphodiesterase of B. miyamotoi. Indigenous relapsing fever should be considered a health concern in Japan.

Ixodes persulcatus is a species of hard tick which is a predominant tick species that spreads a wide array of serious human and animal pathogens. Here, we first assemble the complete mitogenome of I. persulcatus of China. The total length of the mitogenome was 14,539 bp included 36 genes and with a mitogenome structure similar to other ticks. Phylogenetic tree was constructed based on the complete mitogenome of I. persulcatus and closely related 19 species ticks to assess their phylogenic relationship and evolution. We also analyze the differences between the mitogenomes of I. persulcatus of Japan and China. The complete mitogenome data would be useful for further study of I. persulcatus.

The increase in tick-borne encephalitis (TBE) incidence is observed in recent decades in a number of subarctic countries. The reasons of it are widely discussed in scientific publications. The objective of this study was to understand if the climate change in Arkhangelsk Oblast (AO) situated in the north of European subarctic zone of Russia has real impact on the northward expansion of Ixodid ticks and stipulates the increase in TBE incidence. This study analyzes: TBE incidence in AO and throughout Russia, the results of Ixodid ticks collecting in a number of sites in AO, and TBE virus prevalence in those ticks, the data on tick bite incidence in AO, and meteorological data on AO mean annual air temperatures and precipitations.It is established that in recent years TBE incidence in AO tended to increase contrary to its apparent decrease nationwide. In last 10 years, there was nearly 50-fold rise in TBE incidence in AO when compared with 1980–1989. Probably, the increase both in mean annual air temperatures and temperatures during tick active season resulted in the northward expansion of Ixodes Persulcatus, main TBE virus vector. The Ixodid ticks expansion is confirmed both by the results of ticks flagging from the surface vegetation and by the tick bite incidence in the population of AO locations earlier free from ticks. Our mathematical (correlation and regression) analysis of available data revealed a distinct correlation between TBE incidence and the growth of mean annual air temperatures in AO in 1990–2009.Not ruling out other factors, we conclude that climate change contributed much to the TBE incidence increase in AO.

Among main factors that influence intensity of tick-borne encephalitis foci one can distinguish the numbers of core vector (here, taiga tick Ixodes persulcatus) and the percentage of infected ticks. This paper shows the results of five-year monitoring of the tick-borne encephalitis focus, which is situated near Irkutsk city. Detected are the variations in numbers and infestation of the core vector of tick-borne encephalitis, both spatial and temporal. Cause-effect connection between these factors and human TBE morbidity is not found. However, morbidity rates of the Irkutsk population, observed on the annual basis, bear evidence of high activity and intensity of the foci. In this regard, a number of preventive measures is put forward, but further observations concerning all the parameters that influence foci activity are required.