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Tick-borne encephalitis virus (TBEV) has been classified into three subtypes, namely the European (Eu-TBEV), Far Eastern (FE-TBEV), and Siberian (Sib-TBEV). In this study, we discovered a new subtype of TBEV in wild rodent Marmota himalayana in Qinghai-Tibet Plateau in China, proposed as subtype Himalayan (Him-TBEV). Two complete genomes of TBEV were obtained from respiratory samples of 200 marmots. The phylogenetic analysis using the E protein and polyprotein demonstrated that the two strains of Him-TBEV formed an independent branch, separated from Eu-TBEV, Sib-TBEV, and FE-TBEV. The nomenclature of Him-TBEV as a new subtype was also supported by comparative analysis using nucleotide and amino acid sequences of E protein and polyprotein. For E protein, The Him-TBEV showed 82.6-84.6% nucleotide identities and 92.7-95.0% amino acid identities with other three subtypes. For polyprotein, the Him-TBEV showed 83.5-85.2% nucleotide identities and 92.6-94.2% amino acids identities with other three subtypes. Furthermore, of 69 amino acid substitutions profiles detected in complete polyprotein of 112 strains of TBEV, Him-TBEV subtype displayed unique amino acids in the 36 positions. Notably, for the subtype-specific amino acid position 206 of E protein, Him-TBEV shared the Val with Eu-TBEV, but differed from FE-TBEV and Sib-TBEV. The evolutionary analysis with BEAST suggested that Him-TBEV diverged from other subtypes of eastern TBEV group about 2469 years ago. It should be mentioned that Qinghai-Tibet Plateau in China is the plague endemic region where Marmota himalayana is the primary host. The public health significance of discovery of Him-TBEV in Marmota himalayana must be carefully evaluated.

Objectives Tick-borne encephalitis (TBE) is an infection caused by the tick-borne encephalitis virus (TBEV) that can lead to symptoms of central nervous system inflammation. There are five subtypes of TBEV, three of which – European, Siberian and Far Eastern – occur in Europe. As it is thought that different subtype infections exhibit varying clinical courses and outcomes, serological differentiation of the virus subtypes is clearly important. However, to date, this has proved difficult to achieve. Methods An ELISA format was developed based on TBE virus NS1 antigen against the European, Siberian and Far Eastern subtype. The three NS1 antigens were biotechnologically produced in a human cell line and used for ELISA coating. Sera from German (European subtype) and Russian (Siberian and/or Far Eastern subtypes) TBE patients with positive TBEV IgG were used to test the reactivity against these three NS1 antigens. Results Testing of 23 German and 32 Russian TBEV IgG-positive sera showed that the ELISA was able to differentiate between TBEV European subtype and TBEV Siberian and Far Eastern subtype infections. Conclusions In geographical areas where two or more TBEV subtype infections can occur, the NS1-IgG ELISA developed here constitutes an important diagnostic tool to differentiate between European subtype infections and Siberian/Far Eastern subtype infections and to use the new assay for epidemiological studies to clarify the importance of particular subtype infections in an area. Consequently, it may help to better describe and anticipate the clinical courses and outcomes of particular TBEV subtype infections.

The European subtype of tick-borne encephalitis virus (TBEV-Eur; species Orthoflavivirus encephalitidis, family Flaviviridae) was the only tick-borne flavivirus present in central Europe known to cause neurologic disease in humans and several animal species. Here, we report a tick-borne flavivirus isolated from Alpine chamois (Rupicapra rupicapra rupicapra) with encephalitis and attached ticks, present over a wide area in the Alps. Cases were detected in 2017 in Salzburg, Austria, and 2023 in Lombardy and Piedmont, Italy. The virus strains exhibit 94.8–97.3% nucleotide identities to each other and are more closely related to Louping ill viruses (LIV; Orthoflavivirus loupingi; 90–92% identities) than to TBEV-Eur (less than 88%). The chamois-derived virus strains, tentatively termed “Alpine chamois encephalitis virus”, form a well-supported independent genetic clade with Spanish goat encephalitis virus, clearly separated from other LIV. This supports its designation as a new virus subtype with the proposed shared taxonomic name “Spanish goat and Alpine chamois encephalitis virus subtype” within the species Orthoflavivirus loupingi. The zoonotic potential of this newly identified virus subtype as well as its host range in other animal species including farm animals needs to be further investigated.

Tick-borne encephalitis virus (TBEV) is traditionally divided into three main subtypes – European (Eu), Siberian (Sib) and Far Eastern (FE), the distribution of which is confined to the areas of the main vectors, Ixodes ricinus (TBEV-Eu) and Ixodes persulcatus (TBEV-Sib, TBEV-FE). Dermacentor reticulatus also can act as competent vector and participate in TBEV circulation together with the main vectors. It is suggested that there is a specific adaptation not only between TBEV variant and certain tick species, but also between virus variant and local populations of one tick species. In our study, we percoxally infected two populations of I. ricinus, I. persulcatus and D. reticulatus collected in remote areas with three TBEV strains of the main subtypes. Dynamics of the number of TBEV RNA copies and of the number of infectious for mammalian cells virus particles during mono- and mixed infection of ticks were estimated by real-time PCR and plaque assay in PEK cell culture, respectively. Data was obtained that I. ricinus, I. persulcatus and D. reticulatus effectively support the reproduction of TBEV regardless of the strain. Interpopulation differences of local populations of one tick species in the maintenance of TBEV reproduction were revealed in I. persulcatus during mono- and mixed infection and in I. ricinus during mixed infection. Despite minor differences in the level of virus reproduction in ticks, we observed changes in the infectivity of TBEV strains for mammalian cell culture during persistence in different species of ticks. Notably, the TBEV-Eu increased infectivity during adaptation to a non-specific tick species. Thus, we demonstrated that the level of virus reproduction is not the primary factor that determines the adaptation of TBEV to a new tick species. The nature of changes in TBEV infectivity depends on the virus strain and the species of ticks.

Russia is a country with a high incidence of tick-borne encephalitis (TBE). In northwestern regions of Russia, 110 TBE cases were registered in 2021. The largest numbers of TBE cases were registered in the Arkhangelsk region and St. Petersburg. TBEV seropositivity among healthy individuals, including the unvaccinated population in northwestern Russia, was found in 12.2% of studied samples, indicating active TBEV circulation. The prevalence of TBEV is 2.4% in the two tick species most common in northwestern regions of Russia, Ixodes ricinus and Ixodes persulcatus . However, there is still no comprehensive data on the molecular characterization and phylogenetic analysis of the circulating TBEV strains. The purpose of the study was to determine the prevalence of TBEV and to identify its subtypes in ixodid ticks collected in specific areas of northwestern Russian regions. Phylogenetic analysis of E protein sequences of ten obtained strains showed that they all belong to the Siberian subtype, which were clustered into two groups: the most numerous Baltic group, clusteron 3D; and the Vasilchenko group. However, some unique isolates may form new clusterons.

Tick-borne encephalitis virus (TBEV) causes tick-borne encephalitis (TBE), a severe disease of the human central nervous system. Currently, the data on the genetic variants of TBEV in Kyrgyzstan are practically absent. Therefore, the aim of this study was to analyze and describe the genetic diversity of TBEV in this region. The complete genome sequences of seven TBEV strains from the collection of the Scientific Centre for Family Health and Human Reproduction Problems (Irkutsk, Russia) were determined. These strains, isolated from Ixodes persulcatus ticks from Kyrgyzstan, were sequenced using the next generation sequencing approach on a MiSeq high-performance sequencer (Illumina, San Diego, CA, USA). A molecular genetic analysis of the obtained sequences, along with sequences of two previously isolated TBEV strains from Kyrgyzstan available in the GenBank database, demonstrated that the Siberian subtype of three genetic lineages (Zausaev, Vasilchenko and Bosnia) is predominantly distributed in Kyrgyzstan. The Far Eastern subtype of TBEV is also present. To date, this location probably represents the southernmost boundary of these TBEV subtypes’ ranges.

Ixodid ticks are blood-sucking ectoparasites of vertebrates. They constitute an integral part of natural foci and are responsible for the worldwide transmission of infections to humans, which can result in severe symptoms. For instance, the Tomsk region, where three abundant tick species (Dermacentor reticulatus, Ixodes pavlovskyi, I. persulcatus) occur, is an endemic area for tick-borne encephalitis virus (TBEV). An increasing number of novel infectious agents carried by ticks have been identified using metagenomic sequencing. A notable example is the Yezo virus (Orthonairovirus yezoense, YEZV), which was discovered in patients with fever after tick bites in Japan and China between 2014 and 2025. For the first time, we have performed metagenomic sequencing of the virome of ticks collected in the Tomsk region. In a sample obtained from a pool of I. pavlovskyi ticks, all three segments of the YEZV genome were detected. The phylogenetic analysis showed that the newly identified isolate formed a sister group to previously described virus isolates, indicating the presence of a new genetic variant. This study presents the first report of YEZV detection in I. pavlovskyi ticks in the Tomsk region, thereby expanding the geographical range and number of vector species for YEZV and highlighting the importance of monitoring viral agents circulating among ticks in Western Siberia.

(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.

Far Eastern (FE) tick-borne encephalitis virus (TBEV) was thought to be the only endemic subtype in northeastern China. However, in 2024, the Siberian (Sib) subtype of TBEV was found in ticks. This study investigates Sib-TBEV infection in tick-bitten patients in the region. A total of 2573 tick-bitten patients, including 506 inpatients and 2067 outpatients, were enrolled between April and September in both 2023 and 2024 and were tested for Sib-TBEV using real-time quantitative polymerase chain reaction (RT-qPCR). Seven cases were confirmed, with four among inpatients and three among outpatients. All four inpatients exhibited headaches and had hospital stays ranging from 5 to 21 days. Two inpatients with longer hospital stays showed additional symptoms: one experienced dizziness and fatigue, while the other exhibited liver injury and neurological symptoms, including facial paralysis, dysarthria, and meningeal signs. All patients recovered and were discharged without sequelae. Seven Sib-TBEV complete genomes were amplified and classified into the Sib-TBEV Zausaev and Vasilchenko lineages. Bayesian analysis estimated that the most recent common ancestor (tMRCA) of these strains dates to a period between 1898 and 1983, with possible migration pathways from Irkutsk or Zabaykalsky to the Daxing'an Mountains (DXAM) in northeastern China. We identified Sib-TBEV-infected patients in northeastern China and characterized their epidemiological and clinical features. Sib-TBEV may have been circulating in northeastern China for decades, underscoring the need for TBEV surveillance in ticks, animals, and tick-bitten patients.

Tick-borne encephalitis virus (TBEV) is the causative agent of tick-borne encephalitis (TBE), a severe zoonosis occurring in the Palearctic region mainly transmitted through Ixodes ticks. In Italy, TBEV is restricted to the north-eastern part of the country. This report describes for the first time a case of clinical TBE in a roe deer (Capreolus capreolus L.). The case occurred in the Belluno province, Veneto region, an area endemic for TBEV. The affected roe deer showed ataxia, staggering movements, muscle tremors, wide-base stance of the front limbs, repetitive movements of the head, persistent teeth grinding, hypersalivation and prolonged recumbency. An autopsy revealed no significant lesions to explain the neurological signs. TBEV RNA was detected in the brain by real-time RT-PCR, and the nearly complete viral genome (10,897 nucleotides) was sequenced. Phylogenetic analysis of the gene encoding the envelope protein revealed a close relationship to TBEV of the European subtype, and 100% similarity with a partial sequence (520 nucleotides) of a TBEV found in ticks in the bordering Trento province. The histological examination of the midbrain revealed lymphohistiocytic encephalitis, satellitosis and microgliosis, consistent with a viral etiology. Other viral etiologies were ruled out by metagenomic analysis of the brain. This report underlines, for the first time, the occurrence of clinical encephalitic manifestations due to TBEV in a roe deer, suggesting that this pathogen should be included in the frame of differential diagnoses in roe deer with neurologic disease.