Virus Detection in Questing Ticks is not a Sensitive Indicator for Risk Assessment of Tick‐Borne Encephalitis in Humans

Tick‐borne encephalitis virus (TBEV) is the most important tick‐transmitted arbovirus causing human disease in Europe, but information on its endemic occurrence varies between countries because of differences in surveillance systems. Objective data are necessary to ascertain the disease risk for vaccination recommendations and other public health interventions. In two independent, separately planned projects, we used real‐time RT‐PCR to detect TBE virus in questing ticks. In Poland, 32 sampling sites were selected in 10 administrative districts located in regions where sporadic TBE cases were reported. In Germany, 18 sampling sites were selected in two districts located in a region with high TBE incidence. Altogether, >16 000 ticks were tested by real‐time RT‐PCR, with no sample testing positive for TBEV. A systematic search for published studies on TBEV prevalence in ticks in Poland and Germany also suggested that testing large numbers of collected ticks could not consistently assure virus detection in known endemic foci. Although assignment of results to administrative regions is essential for TBE risk mapping, this was possible in only 10 (investigating 22 417 ticks) of 15 published studies (>50 000 ticks) identified. We conclude that the collection and screening of ticks by real‐time RT‐PCR cannot be recommended for assessment of human TBE risk. Alternative methods of environmental TBEV monitoring should be considered, such as serological monitoring of rodents or other wildlife.     

Cervids as sentinel‐species for tick‐borne encephalitis virus in Norway ‐ A serological study

Tick‐borne encephalitis virus (TBEV) is the causative agent of tick‐borne encephalitis (TBE). TBEV is one of the most important neurological pathogens transmitted by tick bites in Europe. The objectives of this study were to investigate the seroprevalence of TBE antibodies in cervids in Norway and the possible emergence of new foci, and furthermore to evaluate if cervids can function as sentinel animals for the distribution of TBEV in the country. Serum samples from 286 moose, 148 roe deer, 140 red deer and 83 reindeer from all over Norway were collected and screened for TBE immunoglobulin G (IgG) antibodies with a modified commercial enzyme‐linked immunosorbent assay (ELISA) and confirmed by TBEV serum neutralisation test (SNT). The overall seroprevalence against the TBEV complex in the cervid specimens from Norway was 4.6%. The highest number of seropositive cervids was found in south‐eastern Norway, but seropositive cervids were also detected in southern‐ and central Norway. Antibodies against TBEV detected by SNT were present in 9.4% of the moose samples, 1.4% in red deer, 0.7% in roe deer, and nil in reindeer. The majority of the positive samples in our study originated from areas where human cases of TBE have been reported in Norway. The study is the first comprehensive screening of cervid species in Norway for antibodies to TBEV, and shows that cervids are useful sentinel animals to indicate TBEV occurrence, as supplement to studies in ticks. Furthermore, the results indicate that TBEV might be spreading northwards in Norway. This information may be of relevance for public health considerations and supports previous findings of TBEV in ticks in Norway.     

COMPARISON BETWEEN PROTON MAGNETIC RESONANCE SPECTROSCOPY FINDINGS IN DOGS WITH TICK‐BORNE ENCEPHALITIS AND CLINICALLY NORMAL DOGS

In vivo diagnosis of tick‐borne encephalitis is difficult due to high seroprevalence and rapid viral clearance, limiting detection of antibodies in blood and cerebrospinal fluid. Magnetic resonance imaging (MRI) characteristics of tick‐borne encephalitis have been reported, however MRI studies can also be negative despite the presence of neurologic signs. Magnetic resonance spectroscopy (¹H MRS) is an imaging method that provides additional information about the metabolic characteristics of brain tissues. The purpose of this retrospective cross‐sectional study was to describe brain metabolites using short echo time single‐voxel ¹H MRS in dogs with confirmed tick‐borne encephalitis and compare them with healthy dogs. Inclusion criteria for the affected dogs were neurological symptoms suggestive of tick‐borne encephalitis, previous endemic stay and tick‐bite, diagnostic quality brain MRI and ¹H MRS studies, and positive antibody titers or confirmation of tick‐borne encephalitis with necropsy. Control dogs were 10, clinically normal beagles that had been used in a previous study. A total of six affected dogs met inclusion criteria. All dogs affected with tick‐borne encephalitis had ¹H MRS metabolite concentration alterations versus control dogs. These changes included mild to moderate decreases in N‐acetyl aspartate and creatine peaks, and mild increases in glutamate/glutamine peaks. No lactate or lipid signal was detected in any dog. Myoinositol and choline signals did not differ between affected and control dogs. In conclusion, findings supported the use of ¹H MRS as an adjunctive imaging method for dogs with suspected tick‐borne encephalitis and inconclusive conventional MRI findings.     

Serological Investigations of Red Foxes (Vulpes vulpes L.) for Determination of the Spread of Tick‐borne Encephalitis in Northrhine‐Westphalia

Serum samples from 786 red foxes shot between January 1995 and August 1996 in the southern half of Northrhine‐Westphalia, located in western Germany, were tested for the presence of antibodies against tick‐borne encephalitis (TBE) virus using the Immunozym FSME IgG All Species‐ELISA^® (Immuno, Heidelberg, Germany) as a screening test: 759 sera were negative, 23 (2.9 %) were borderline, and four (0.5 %) were positive. Nine of the 27 ELISA reactive sera were confirmed by the TBE Western‐Blot (Immuno, Heidelberg, Germany). Furthermore these 27 sera were tested for neutralizing antibodies by means of a plaque reduction neutralization test (PRNT) against TBE and West Nile viruses. Only one single serum was found to have a neutralization titre (+ 1 : 800 PRNT₈₀) against TBE virus. All other 26 sera were negative for neutralizing antibodies against TBE or West Nile virus. Since the titre of the single serum is low, it can be interpreted that if TBE virus is present, its prevalence is extremely low. Northrhine‐Westphalia is not classified as a TBE‐endemic area. Further calculated serological testing of game and virological investigation of collected ticks in the affected area seem to be meaningful and necessary.     

Public Health and Vector‐Borne Diseases – A New Concept for Risk Governance

Public Health is defined as an interdisciplinary multilevel approach that deals with questions of preventing diseases at the population level. In this context, this paper focuses on vector‐borne diseases as an important threat with an increasing impact on human and animal health. Emphasis is laid on an integrated health approach (‘One‐Health’ initiative) as it recognizes the interrelated nature of both human and animal health. The importance of vector‐borne diseases to new and emerging diseases in Europe was demonstrated, for example, by the recent outbreak of West Nile virus infections in Greece, Northern Italy and Hungary; the spread of Crimean‐Congo haemorrhagic fever virus across Turkey, south‐western countries of the former USSR and the Balkans; the dramatic increase in hantavirus infections in Germany in 2012; and the dengue virus outbreak in Portugal in the same year. This paper provides a systematic approach for the analysis, assessment and governance of emerging health risks attributed to vector‐borne diseases by using a holistic approach developed by the International Risk Governance Council (IRGC), called the ‘IRGC Risk Governance Framework’. It can be used by decision‐makers and general Public Health authorities in order to evaluate the situation regarding any specific pathogen or Public Health risk and to decide if additional measures should be implemented.     

THREE TESLA MAGNETIC RESONANCE IMAGING FINDINGS IN 12 CASES OF CANINE CENTRAL EUROPEAN TICK‐BORNE MENINGOENCEPHALOMYELITIS

Central European tick‐borne encephalomyelitis can be challenging to diagnose in dogs because the virus may not be detected in blood and cerebrospinal fluid (CSF) after the first viremic stage of the disease. The purpose of this retrospective case series study was to describe 3 Tesla magnetic resonance imaging (3T MRI) findings in a sample of dogs with a confirmed diagnosis of tick‐borne encephalomyelitis. Dogs were included if they had neurological signs consistent with tick‐borne encephalomyelitis, history of a stay in endemic areas for tick‐borne encephalomyelitis virus, 3T MRI of the brain and/or spinal cord, cerebrospinal fluid changes compatible with viral infection and positive antibody titers in cerebrospinal fluid or pathologic confirmation of tick‐borne encephalomyelitis. Twelve dogs met inclusion criteria. Ten out of 12 patients had 3T MRI lesions at the time of presentation. One patient had persistent lesions in follow‐up MRI. The 3T MRI findings included bilateral and symmetrical gray matter distributed lesions involving the thalamus, hippocampus, brain stem, basal nuclei, and ventral horn on the spinal cord. All lesions were hyperintense in T2‐weighted sequences compared to white matter, iso‐ to hypointense in T1‐weighted, nonenhancing, and had minimal or no mass effect or perilesional edema. Six patients survived while the remaining six dogs were euthanized. Necropsy revealed neuronophagia and gliosis of the gray matter of the affected regions seen in 3T MRI, in addition to the cerebellum. Findings from the current study indicated that tick‐borne encephalomyelitis should be included in the differential diagnosis list for dogs with the above described 3T MRI characteristics.     

Following a tick bite: double infections by tick-borne encephalitis virus and the spirochete Borrelia and other potential multiple infections

In Central Europe and large parts of Asia, tick-borne-encephalitis (TBE) and Lyme borreliosis caused by the spirochetal bacterium of the genus Borrelia are among the most common diseases transmitted by the bite of a tick. When in regions with overlapping TBE virus and Borrelia endemicity, a tick bite causes the victim to become ill, it is important that appropriate serological and other laboratory investigations form part of the differential diagnosis. Account must always be taken of the fact that a tick bite may be followed by a double infection with the TBE virus and Borrelia. For this reason, a comprehensive diagnostic work-up aimed at detecting co-infection by both pathogens, even when the tick bite occurs in an endemic region for both pathogens but the initial clinical symptoms suggest an infection with only one of the two pathogens. The present article discusses a number of published cases of a co-infection with TBE virus and Borrelia and other potential multiple infections.     

Evidence of Powassan/deer tick virus in adult black‐legged ticks (Ixodes scapularis) recovered from hunter‐harvested white‐tailed deer (Odocoileus virginianus) in Pennsylvania: A public health perspective

Studies reporting tick infection rates for Powassan virus (POWV), an emerging zoonotic arthropod‐borne pathogen responsible for POWV disease in the Commonwealth of Pennsylvania, are limited. To determine the presence and ascertain a statewide prevalence of POWV, ticks were collected from 9,912 hunter‐harvested white‐tailed deer (Odocoileus virginianus) heads presented to six regional Pennsylvania Game Commission Chronic Wasting Disease sampling stations in early December of 2013, 2014 and 2015. Of the 2,973 ticks recovered, 1,990 (66.9%) were identified as adult Ixodes scapularis (black‐legged tick). The 1,990 I. scapularis ticks were PCR‐tested for the presence of POWV. The ticks had a statewide Powassan/deer tick virus infection rate of 0.05%, providing evidence of this pathogen in Pennsylvania's adult I. scapularis ticks and supporting the need for more comprehensive pathogen prevalence assessment strategies, as well as increased public health awareness for this emerging zoonotic arthropod‐borne pathogen of public health concern.     

Toscana virus (TOSV) exposure is confirmed in blood donors from Central, North and South/Southeast Anatolia, Turkey

Toscana virus (TOSV), a sandfly fever virus serotype of medical and public health importance, is a major pathogen involved in aseptic meningtis occurring in Mediterranean countries and poses a threat to the residents as well as travellers. Limited data on TOSV activity are present from Turkey despite being located in the endemic zone. We aimed to identify TOSV exposure in 1115 healthy blood donors at the Hacettepe University Hospital Blood Bank in Ankara, Turkey, using commercial indirect fluorescence assays (IFAs) and virus neutralization test (VNT) for antibody detection and specificity confirmation. A total of 199 samples (17.8%) were positive for anti-TOSV that include IgG reactivity in 10.4%, IgM reactivity in 8.2% and IgM + IgG reactivity in 0.7% of the sera. Anti-TOSV specificity could be confirmed via VNT in 56% of the IgG- and 43.6% of the IgM-positive sera, making up a total of 58 samples (5.2%). Risk factors associated with TOSV IgG reactivity were male gender, residing in rural areas, frequent sighting of mosquitoes/sandflies and working outdoors. TOSV-specific antibody prevalence increased significantly with age. Evidence of exposure to other sandfly fever viruses was noted. These data reveal that mild or asymptomatic infections with TOSV are frequent in central and northern Anatolia. TOSV exposure has also been identified in residents of 9 provinces in southern/southeastern Anatolia for the first time.     

Geographical distribution and prevalence of tick‐borne encephalitis virus in questing Ixodes ricinus ticks and phylogeographic structure of the Ixodes ricinus vector in Norway

The tick‐borne encephalitis virus (TBEV), a zoonotic flaviviral infection, is endemic in large parts of Norway and Eurasia. Humans are mainly infected with TBEV via bites from infected ticks. In Norway, the main geographical distribution of ticks is along the Norwegian coastline from southeast (~59°N) and up to the southern parts of Nordland County (~65°N). In this study, we collected ticks by flagging along the coast from Østfold County to Nordland County. By whole‐genome sequencing of the mitochondrial genome of Ixodes ricinus, the phylogenetic tree suggests that there is limited phylogeographic structure both in Norway and in Europe. The overall TBEV prevalence is 0.3% for nymphs and 4.3% for adults. The highest estimated TBEV prevalence in adult ticks was detected in Rogaland and Vestfold County, while for nymphs it is highest in Vestfold, Vest‐Agder and Rogaland. The present work is one of the largest studies on distribution and prevalence of TBEV in ticks in Scandinavia, showing that the virus is wider distributed in Norway than previously anticipated.     

Tick-borne encephalitis virus as a possible cause of optic neuritis in a dog

A 3-year-old spayed female Siberian Husky was presented due to acute vision loss. Examination revealed bilateral optic neuritis and lymphocytic meningoencephalitis. The serum (1:800) and cerebrospinal fluid (CSF; 1:200) immunoglobulin (Ig)G titers for tick-borne encephalitis virus (TBEV) were elevated as were the serum IgG titer for Anaplasma phagocytophilum (1:640) and serum IgM titer for Toxoplasma gondii (1:20). Intracytoplasmic inclusion bodies such as ehrlichial or anaplasmal morulae were not observed in the CSF or blood smear. The dog was treated with methylprednisone and doxycycline. The left eye regained vision; the right eye remained blind. Anti-inflammatory therapy was stopped on day 18 after diagnosis. Four days later the dog showed evidence of hyperesthesia in the cervical region. Analysis of CSF showed no abnormalities and CSF IgG titers for TBEV and A. phagocytophilum were negative. Funduscopic evidence of active papillitis was absent on day 22 in the left eye and on day 86 in the right eye. On day 243, the dog was presented again with lethargy, ataxia, disorientation and temporary head tilt. The IgG titer for TBEV was again elevated in the CSF (1:800) and in serum (1:400). After interpretation of all findings, we assume that meningoencephalitis and optic neuritis in this patient was caused by TBEV and associated immune-mediated inflammation. In endemic areas, TBEV should be considered as cause of optic neuritis in dogs.     

Epizootiologic background of dissimilar distribution of human cases of Lyme borreliosis and tick-borne encephalitis in a joint endemic area

Lyme borreliosis (LB) and tick-borne encephalitis (TBE) are co-endemic in some parts of Europe, however, their distribution differs despite a common tick vector and comparable animal hosts. A serosurvey of game and small mammals was made in a highly endemic area and compared with historical data in human cases; the epidemiologic risk and the population density of game were modelled using a geographic information system. While LB-risk corresponded with an overall population density of game (red deer, roe deer, mouflon, wild boar) regardless of mouse abundance, TBE-risk suggested a dependence on the abundance of mice on the one hand, and game, particularly roe deer, on the other. While the prevalence of TBE-antibodies generally grew with the game’s age, it was virtually constant at about 65% in LB irrespective of species. It implies a cumulation of scarce TBE-infection histories during the game’s lifetime, and thus a limited size of TBE-foci relative to the living space of these animals, as well as omnipresent LB-foci, in which the animals became continuously re-infected. More ecological prerequisites seem necessary to keep TBE circulating in nature which may be responsible for its confined distribution.     

Tick bite and Lyme disease‐related emergency department encounters in New Hampshire, 2010–2014

Lyme disease (LD) is a common tick‐borne disease in New Hampshire (NH). While LD is a reportable condition and cases are counted for public health surveillance, many more people receive care for tick bites or diagnoses of LD than are reflected in surveillance data. NH's emergency department (ED) data system was queried for tick bite and LD‐related encounters. Chief complaint text was queried for words related to LD or tick bites. International Classification of Diseases 9th Revision (ICD‐9) codes were queried for the LD diagnosis code (088.81). Emergency department patient data were matched to reportable disease data to determine the proportion of ED patients reported to the health department as a suspected LD case. Data were analysed to calculate frequencies for key demographic and reporting characteristics. From 2010 to 2014, 13,615 tick bite or LD‐related ED encounters were identified in NH, with most due to tick bites (76%). Of 3,256 patients with a LD‐related ED encounter, 738 (23%) were reported to the health department as a suspected LD case. The geographic distribution of ED patients was similar to reported LD cases; however, the regions of the state that experienced higher rates of ED encounters were different than the regions that observed higher rates of reported LD cases. Seasonal distribution of ED encounters peaked earlier than reported LD cases with a second peak in the fall. While age and sex distribution was similar among ED patients and reported LD cases, the rates for children 5 years and younger and adults 65 years and older were greater for ED encounters. Patients frequently visit the ED to seek care for tick bites and suspected LD. Results of ED data analyses can be used to target education, in particular for ED providers and the public through timely distribution of evidence‐based educational materials and training programmes.     

Tick-borne encephalitis epidemic in Hungary 1951–2021: The story and lessons learned

The authors analysed epidemiological data of the Hungarian tick-borne encephalitis epidemic from the past seven decades. A total of 911 meningitis serosa cases were described from 1930-1950 s by local hospital physicians, indicating that the virus had been present in the country decades before its official identification in 1952. The virus spread freely in the 1950s–1960s, occupying almost all habitats where ticks occurred in large numbers. The increasing number of cases drove authorities to classify this illness as a notifiable disease in 1977 and to organize the first measures to stop the epidemic. Statistical analysis revealed that the large-scale vaccination launched from the 1990s was responsible for the sharp decrease in the number of human cases from 1997. A significant negative correlation was found between the number of vaccine doses sold and human cases 6 years later. The TBEV endemic area covers 16.57% of the territory and 16.65% of the population of the country. In the last 10 years, 186,000 vaccine doses/year in average were enough to keep the incidence of human TBEV infections between 0.45 and 0.06/100,000 persons. A 20-year-long study found evidence for easing clinical signs in TBEV-infected hospitalized patients. Statistics found a sharp decrease in the number of samples sent for TBEV diagnosis after 1989. Male dominance of patients was characteristic of the epidemics since the 1940s, but now analysis of detailed data from the 1981–2021 period (60.5%–87.5%) proved the statistical significance of this dominance. Obviously, the voluntary vaccination programme was the tool which broke the spread of the epidemic. Widespread public awareness of the disease and the tick vector, probable evolutionary spread of less pathogenic virus strains supplemented with the vaccination campaign led to a negligible level of human TBE cases in Hungary in the last years.     

Discrepancies between self‐reported tick bites and evidence of tick‐borne disease exposure among nomadic Mongolian herders

Twenty‐six per cent of Mongolians live pastoral lifestyles, increasing their likelihood of exposure to ticks and placing them at a higher risk for contracting tick‐borne diseases (TBDs). Anaplasma spp. and Rickettsia spp. have been identified in ticks, livestock and humans in Mongolia, but no known qualitative research has been conducted investigating the association between nomadic herder characteristics, tick bite history and exposure to TBDs. To better understand the association between self‐reported tick bites and symptoms versus actual exposure to TBDs, this study paired serological data with 335 surveys administered to Mongolian herders, ages 12–69, from 2014 to 2015. Logistic regression results identified no significant associations between reported tick bites or symptoms with serological evidence of Anaplasma spp. and Rickettsia spp. controlling for age, gender and aimag. Among the 335 respondents who were seropositive to either Anaplasma spp. or Rickettsia spp., 32.9% self‐reported experiencing abnormal symptoms such as redness, inflammation, headache, arthritis or fever after being bitten. Alternatively, 17.3% (58/335) of individuals reported experiencing symptoms following a tick bite in instances where serological results indicated no exposure to Anaplasma spp. or Rickettsia spp. Results also identified inconsistencies in reporting and seroprevalence among different age groups, with children having the highest reporting and treatment seeking rates but low levels of exposure in comparison with other groups. While survey results showed that individuals were aware of peak tick seasons and tick species that inhabit specific areas, 58% of heads of households (49/84) were unaware that ticks can cause disease in livestock or dogs. This study suggests that herders are an at‐risk population in Mongolia with gaps in awareness of TBD risk. Increased surveillance paired with focused outreach to prevent TBDs targeted to the herder population is encouraged.     

Update on Canine and Feline Blood Donor Screening for Blood‐Borne Pathogens

An update on the 2005 American College of Veterinary Internal Medicine (ACVIM) Consensus Statement on blood donor infectious disease screening was presented at the 2015 ACVIM Forum in Indianapolis, Indiana, followed by panel and audience discussion. The updated consensus statement is presented below. The consensus statement aims to provide guidance on appropriate blood‐borne pathogen testing for canine and feline blood donors in North America.     

Transmission ecology of rodent‐borne diseases: New frontiers

Rodents are recognized reservoir hosts for many human zoonotic pathogens. The current trends resulting from anthropocene defaunation suggest that in the future they, along with other small mammals, are likely to become the dominant mammals in almost all human‐modified environments. Recent intricate studies on bat‐borne emerging diseases have highlighted that many gaps exist in our understanding of the zoonotic transmission of rodent‐borne pathogens. This has emphasized the need for scientists interested in rodent‐borne diseases to integrate rodent ecology into their analysis of rodent‐borne pathogen transmission in order to identify in more detail the mechanisms of spillover and chains of transmission. Further studies are required to better understand the true impact of rodent abundance and the importance of pathogen sharing and circulation in multi‐host– multi‐pathogen communities. We also need to explore in more depth the roles of generalist and abundant species as the potential links between pathogen‐sharing, co‐infections and disease transmission.     

Seroepidemiology and genotyping of hepatitis E virus in Singapore reveal rise in number of cases and similarity of human strains to those detected in pig livers

Hepatitis E virus (HEV) causes 20 million infections worldwide yearly, of which only about 3.3 million are symptomatic. In developed Asian countries, HEV strains detected in human sera and in food sources were genetically similar, suggesting that indigenous HEV infections may be largely food‐borne. To assess the burden of hepatitis E in Singapore, we performed a seroepidemiologic study of the infection. Additionally, we carried out HEV genotyping on archived, residual HEV IgM‐positive serum samples collected between 2014 and 2016 (n = 449), and on pig liver samples (n = 36) purchased from wet markets and supermarkets. Our study shows a rise in hepatitis E incidence (IgM) from 1.7 to 4.1 cases per 100,000 resident population from 2012 to 2016 and an increase in hepatitis E IgG positivity rate among residents from 14% in 2007 to 35% in 2016. Other findings also suggest the epidemiology of hepatitis E in Singapore has shifted, from it being mainly a disease imported from the Indian subcontinent, to one that is now increasingly prevalent in our resident population. Genotypes obtained from 143 human samples identified the majority to be genotype 3 (n = 121), 21 to be genotype 1 and one to be genotype 4. Further phylogenetic analyses suggest genotype 3a to be the cause of indigenous infections in residents, which showed genetic similarity to the genotype 3a strains detected in pig livers. This link between the strains in the majority of human samples and those in pig livers consumed by the public suggests a possible food‐borne route of HEV infection in Singapore.     

Ongoing Activity of Toscana Virus Genotype A and West Nile Virus Lineage 1 Strains in Turkey: A Clinical and Field Survey

Toscana virus (TOSV), West Nile virus (WNV) and tickborne encephalitis virus (TBEV) are among major viral pathogens causing febrile disease and meningitis/encephalitis. The impact of these viruses was investigated at a referral centre in Ankara Province, Central Anatolia in 2012, where previous reports suggested virus circulation but with scarce information on clinical cases and vector activity. Serum and/or cerebrospinal fluid samples from 94 individuals were evaluated, in addition to field‐collected arthropod specimens that included 767 sandflies and 239 mosquitoes. Viral nucleic acids in clinical samples and arthropods were sought via specific and generic nested/real‐time PCRs, and antibody responses in clinical samples were investigated via commercial indirect immunofluorescence tests (IIFTs) and virus neutralization. A WNV antigen assay was also employed for mosquitoes. WNV neuroinvasive disease has been identified in a 63‐year‐old male via RNA detection, and the WNV strain was characterized as lineage 1. TOSV infections were diagnosed in six individuals (6.3%) via RNA or IgM detection. Partial sequences in a 23‐year‐old female, presented with fever and transient pancytopenia, were characterized as TOSV genotype A. Febrile disease with arthralgia and/or peripheral cranial nerve involvement was noted in cases with TOSV infections. Previous WNV and TOSV exposures have been observed in 5.3% and 2.1% of the subjects, respectively. No confirmed TBEV exposure could be identified. Morphological identification of the field‐collected mosquitoes revealed Culex pipiens sensu lato (74.4%), Anopheles maculipennis (20.9%), An. claviger (2.1%) and others. Sandfly species were determined as Phlebotomus papatasi (36.2%), P. halepensis (27.3%), P. major s. l. (19.3%), P. sergenti (8.9%), P. perfiliewi (4.4%), P. simici (2.6%) and others. Viral infections in arthropods could not be demonstrated. TOSV genotype A and WNV lineage 1 activity have been demonstrated as well as serologically proven exposure in patients. Presence of sandfly and mosquito species capable of virus transmission has also been revealed.