Possible expansion of Ixodes ricinus in the United Kingdom identified through the Tick Surveillance Scheme between 2013 and 2020.

The tick Ixodes ricinus (Ixodida: Ixodidae, Linnaeus) is the main vector of several pathogens including Borrelia burgdorferi s.l. (agent of Lyme borreliosis) and tick-borne encephalitis virus. Its distribution depends on many factors including suitable habitat, climate and presence of hosts. In this study, we present records of I. ricinus bites on humans, dogs (Canis lupus familiaris; Carnivora: Canidae, L.) and cats (Felis catus; Carnivora: Felidiae, L.) in the United Kingdom (UK) obtained through the Tick Surveillance Scheme between 2013 and 2020. We divided the UK into 20 km x 20 km grids and 9.2% (range 1.2%-30%) of grids had at least one record every year since 2013. Most regions reported a yearly increase in the percentage of grids reporting I. ricinus since 2013 and the highest changes occurred in the South and East England with 5%-6.7% of new grids reporting I. ricinus bites each year in areas that never reported ticks before. Spatiotemporal analyses suggested that, while all regions recorded I. ricinus in new areas every year, there was a yearly decline in the percentage of new areas covered, except for Scotland. We discuss potential drivers of tick expansion, including reforestation and increase in deer populations.

Mapping and monitoring tick (Acari, Ixodida) distribution, seasonality, and host associations in the United Kingdom between 2017 and 2020.

Tick-borne disease risk is intrinsically linked to the distribution of tick vector species. To assess risk and anticipate disease emergence, an understanding of tick distribution, host associations, and seasonality is needed. This can be achieved, to some extent, using passive surveillance supported by engagement with the public, animal health, and public health experts. The Tick Surveillance Scheme (TSS) collects data and maps tick distribution across the United Kingdom (UK). Between 2017 and 2020, 3720 tick records were received and 39 tick species were detected. Most records were acquired in the UK, with a subset associated with recent overseas travel. The dominant UK acquired species was Ixodes ricinus (Ixodida: Ixodidae, Linnaeus), the main vector of Lyme borreliosis. Records peaked during May and June, highlighting a key risk period for tick bites. Other key UK species were detected, including Dermacentor reticulatus (Ixodida: Ixodidae, Fabricius) and Haemaphysalis punctata (Ixodida: Ixodidae, Canestrini & Fanzago) as well as several rarer species that may present novel tick-borne disease risk to humans and other animals. Updated tick distribution maps highlight areas in the UK where tick exposure has occurred. There is evidence of increasing human tick exposure over time, including during the COVID-19 pandemic, but seasonal patterns remain unchanged.

The Genetic Diversity of Rickettsiella Symbionts in Ixodes ricinus Throughout Europe.

Rickettsiella species are bacterial symbionts that are present in a great variety of arthropod species, including ixodid ticks. However, little is known about their genetic diversity and distribution in Ixodes ricinus, as well as their relationship with other tick-associated bacteria. In this study, we investigated the occurrence and the genetic diversity of Rickettsiella spp. in I. ricinus throughout Europe and evaluated any preferential and antagonistic associations with Candidatus Midichloria mitochondrii and the pathogens Borrelia burgdorferi sensu lato and Borrelia miyamotoi. Rickettsiella spp. were detected in most I. ricinus populations investigated, encompassing a wide array of climate types and environments. The infection prevalence significantly differed between geographic locations and was significantly higher in adults than in immature life stages. Phylogenetic investigations and protein characterization disclosed four Rickettsiella clades (I-IV). Close phylogenetic relations were observed between Rickettsiella strains of I. ricinus and other arthropod species. Isolation patterns were detected for Clades II and IV, which were restricted to specific geographic areas. Lastly, although coinfections occurred, we did not detect significant associations between Rickettsiella spp. and the other tick-associated bacteria investigated. Our results suggest that Rickettsiella spp. are a genetically and biologically diverse facultative symbiont of I. ricinus and that their distribution among tick populations could be influenced by environmental components.

Spatial and temporal heterogeneity of the density of Borrelia burgdorferi-infected Ixodes ricinus ticks across a landscape: A 5-year study in southern England.

The density of Borrelia burgdorferi-infected Ixodes ricinus nymphs (DIN) was investigated during 2013-2017 across a Lyme disease-endemic landscape in southern England. The density of nymphs (DON), nymph infection prevalence (NIP), and DIN varied across five different natural habitats, with the highest DIN in woodland edge and high biodiversity woodlands. DIN was significantly lower in scrub grassland compared to the woodland edge, with low DON and no evidence of infection in ticks in non-scrub grassland. Over the 5 years, DON, NIP and DIN were comparable within habitats, except in 2014, with NIP varying three-fold and DIN significantly lower compared to 2015-2017. Borrelia garinii was most common, with bird-associated Borrelia (B. garinii/valaisiana) accounting for ~70% of all typed sequences. Borrelia burgdorferi sensu stricto was more common than B. afzelii. Borrelia afzelii was more common in scrub grassland than woodland and absent in some years. The possible impact of scrub on grazed grassland, management of ecotonal woodland margins with public access, and the possible role of birds/gamebirds impacting NIP are discussed. Mean NIP was 7.6%, highlighting the potential risk posed by B. burgdorferi in this endemic area. There is a need for continued research to understand its complex ecology and identify strategies for minimizing risk to public health, through habitat/game management and public awareness.

Ixodes ricinus and Borrelia burgdorferi sensu lato in the Royal Parks of London, UK.

Assessing the risk of tick-borne disease in areas with high visitor numbers is important from a public health perspective. Evidence suggests that tick presence, density, infection prevalence and the density of infected ticks can vary between habitats within urban green space, suggesting that the risk of Lyme borreliosis transmission can also vary. This study assessed nymph density, Borrelia prevalence and the density of infected nymphs across a range of habitat types in nine parks in London which receive millions of visitors each year. Ixodes ricinus were found in only two of the nine locations sampled, and here they were found in all types of habitat surveyed. Established I. ricinus populations were identified in the two largest parks, both of which had resident free-roaming deer populations. Highest densities of nymphs (15.68 per 100 m2) and infected nymphs (1.22 per 100 m2) were associated with woodland and under canopy habitats in Richmond Park, but ticks infected with Borrelia were found across all habitat types surveyed. Nymphs infected with Borrelia (7.9%) were only reported from Richmond Park, where Borrelia burgdorferi sensu stricto and Borrelia afzelii were identified as the dominant genospecies. Areas with short grass appeared to be less suitable for ticks and maintaining short grass in high footfall areas could be a good strategy for reducing the risk of Lyme borreliosis transmission to humans in such settings. In areas where this would create conflict with existing practices which aim to improve and/or meet historic landscape, biodiversity and public access goals, promoting public health awareness of tick-borne disease risks could also be utilised.

Tick-Borne Encephalitis Virus, United Kingdom.

During February 2018-January 2019, we conducted large-scale surveillance for the presence and prevalence of tick-borne encephalitis virus (TBEV) and louping ill virus (LIV) in sentinel animals and ticks in the United Kingdom. Serum was collected from 1,309 deer culled across England and Scotland. Overall, 4% of samples were ELISA-positive for the TBEV serocomplex. A focus in the Thetford Forest area had the highest proportion (47.7%) of seropositive samples. Ticks collected from culled deer within seropositive regions were tested for viral RNA; 5 of 2,041 ticks tested positive by LIV/TBEV real-time reverse transcription PCR, all from within the Thetford Forest area. From 1 tick, we identified a full-length genomic sequence of TBEV. Thus, using deer as sentinels revealed a potential TBEV focus in the United Kingdom. This detection of TBEV genomic sequence in UK ticks has important public health implications, especially for undiagnosed encephalitis.

A probable case of tick-borne encephalitis (TBE) acquired in England, July 2019.

The United Kingdom (UK) has thus far been considered to be free from tick-borne encephalitis (TBE), yet in July 2019, a German infant developed serologically diagnosed TBE following a tick bite in southern England. This first report of a probable human case together with recent findings of TBE virus in ticks in foci in England suggest that TBE may be acquired in parts of England and should be considered in patients with aetiologically-unexplained neurological manifestations.

Detection of new endemic focus of tick-borne encephalitis virus (TBEV), Hampshire/Dorset border, England, September 2019.

The presence of tick-borne encephalitis virus (TBEV) was detected in a questing tick pool in southern England in September 2019. Hitherto, TBEV had only been detected in a limited area in eastern England. This southern English viral genome sequence is distinct from TBEV-UK, being most similar to TBEV-NL. The new location of TBEV presence highlights that the diagnosis of tick-borne encephalitis should be considered in encephalitic patients in areas of the United Kingdom outside eastern England.

Risk factors for tick attachment in companion animals in Great Britain: a spatiotemporal analysis covering 2014-2021.

BACKGROUND: Ticks are an important driver of veterinary health care, causing irritation and sometimes infection to their hosts. We explored epidemiological and geo-referenced data from > 7 million electronic health records (EHRs) from cats and dogs collected by the Small Animal Veterinary Surveillance Network (SAVSNET) in Great Britain (GB) between 2014 and 2021 to assess the factors affecting tick attachment in an individual and at a spatiotemporal level. METHODS: EHRs in which ticks were mentioned were identified by text mining; domain experts confirmed those with ticks on the animal. Tick presence/absence records were overlaid with a spatiotemporal series of climate, environment, anthropogenic and host distribution factors to produce a spatiotemporal regression matrix. An ensemble machine learning spatiotemporal model was used to fine-tune hyperparameters for Random Forest, Gradient-boosted Trees and Generalized Linear Model regression algorithms, which were then used to produce a final ensemble meta-learner to predict the probability of tick attachment across GB at a monthly interval and averaged long-term through 2014-2021 at a spatial resolution of 1 km. Individual host factors associated with tick attachment were also assessed by conditional logistic regression on a matched case-control dataset. RESULTS: In total, 11,741 consultations were identified in which a tick was recorded. The frequency of tick records was low (0.16% EHRs), suggesting an underestimation of risk. That said, increased odds for tick attachment in cats and dogs were associated with younger adult ages, longer coat length, crossbreeds and unclassified breeds. In cats, males and entire animals had significantly increased odds of recorded tick attachment. The key variables controlling the spatiotemporal risk for tick attachment were climatic (precipitation and temperature) and vegetation type (Enhanced Vegetation Index). Suitable areas for tick attachment were predicted across GB, especially in forests and grassland areas, mainly during summer, particularly in June. CONCLUSIONS: Our results can inform targeted health messages to owners and veterinary practitioners, identifying those animals, seasons and areas of higher risk for tick attachment and allowing for more tailored prophylaxis to reduce tick burden, inappropriate parasiticide treatment and potentially TBDs in companion animals and humans. Sentinel networks like SAVSNET represent a novel complementary data source to improve our understanding of tick attachment risk for companion animals and as a proxy of risk to humans.

Field-based assessments of the seasonality of Culex pipiens sensu lato in England: an important enzootic vector of Usutu and West Nile viruses.

BACKGROUND: Usutu virus (USUV), which is closely related to West Nile virus (WNV), sharing a similar ecology and transmission cycle, was first reported in the UK in the southeast of England in 2020. Both USUV and WNV are emerging zoonotic viruses hosted by wild birds. The 2020 finding of USUV in England raised awareness of this virus and highlighted the importance of understanding the seasonality of Culex pipiens sensu lato (Cx. pipiens s.l.), the main enzootic vector of these viruses. Zoos are prime locations for trapping mosquitoes because of their infrastructure, security, and range of vertebrate hosts and aquatic habitats. METHODS: Three independent zoo-based case studies at four locations that cover the seasonality of Cx. pipiens s.l. in England were undertaken: (i) London Zoo (Zoological Society London [ZSL]) and surrounding areas, London; (ii) Chester Zoo (Cheshire); (ii) Twycross Zoo (Leicestershire); and (iv) Flamingo Land (zoo; North Yorkshire). Various adult mosquito traps were used to catch adult Cx. pipiens s.l. across seasons. RESULTS: High yields of Cx. pipiens s.l./Culex torrentium were observed in Biogents-Mosquitaire and Center for Disease Control and Prevention Gravid traps in all studies where these traps were used. Mosquito counts varied between sites and between years. Observations of adult Cx. pipiens s.l./Cx. torrentium abundance and modelling studies demonstrated peak adult abundance between late July and early August, with active adult female Cx. pipiens s.l./Cx. torrentium populations between May and September. CONCLUSIONS: The information collated in this study illustrates the value of multiple mosquito monitoring approaches in zoos to describe the seasonality of this UK vector across multiple sites in England and provides a framework that can be used for ongoing and future surveillance programmes and disease risk management strategies.

Modelling the current and future temperature suitability of the UK for the vector Hyalomma marginatum (Acari: Ixodidae).

Hyalomma marginatum is the main vector of Crimean-Congo haemorrhagic fever virus (CCHFV) and spotted fever rickettsiae in Europe. The distribution of H. marginatum is currently restricted to parts of southern Europe, northern Africa and Asia, and one of the drivers limiting distribution is climate, particularly temperature. As temperatures rise with climate change, parts of northern Europe currently considered too cold for H. marginatum to be able to survive may become suitable, including the United Kingdom (UK), presenting a potential public health concern. Here we use a series of modelling methodologies to understand whether mean air temperatures across the UK during 2000-2019 were sufficient for H. marginatum nymphs to moult into adult stages and be able to overwinter in the UK if they were introduced on migratory birds. We then used UK-specific climate projections (UKCP18) to determine whether predicted temperatures would be sufficient to allow survival in future. We found that spring temperatures in parts of the UK during 2000-2019 were warm enough for predicted moulting to occur, but in all years except 2006, temperatures during September to December were too cold for overwintering to occur. Our analysis of the projections data suggests that whilst temperatures in the UK during September to December will increase in future, they are likely to remain below the threshold required for H. marginatum populations to become established.

Ixodes ricinus density, Borrelia prevalence and the density of infected nymphs along an urban-rural gradient in southern England.

Ticks are found across a range of habitats, with woodland being particularly important for high densities and prevalence of Borrelia infection. Assessments of risk in urban woodland can be difficult if there are low densities and small sample sizes for Borrelia prevalence estimates. This study targeted six urban woodlands with established tick populations, as well as six woodlands in peri-urban zones and six woodlands in rural zones in and around the cities of Bath and Southampton, in the South of England. Nymph densities were estimated, and 100 nymphs were tested from each of the 18 woodlands studied. Ixodes ricinus ticks were found in all woodlands surveyed, and overall density of nymphs (DON) per 100 m2 was 18.17 in urban woodlands, 26.0 in peri-urban woodlands and 17.67 in rural woodlands. Out of 600 nymphs tested across urban woodlands, 10.3% were infected with Borrelia. The same proportion of nymphs collected in rural woodlands were positive for Borrelia. In peri-urban woodlands, 10.8% of nymphs tested positive. Across both cities combined, density of infected nymphs (DIN) was 2.73 per 100 m2 in peri-urban woodland, 1.87 per 100 m2 in urban woodland and 1.82 per 100 m2 in rural woodland. Overall, DON, Borrelia prevalence and DIN did not differ significantly along an urban-rural gradient. This suggests the risk of Lyme borreliosis transmission could be similar, or perhaps even elevated in urban woodland if there is higher public footfall, subsequent contact with ticks and less awareness of the risks. This is particularly important from a public health perspective, as Borrelia garinii dominated across the gradient and this genospecies is linked to neuroborreliosis.

Impact of green space connectivity on urban tick presence, density and Borrelia infected ticks in different habitats and seasons in three cities in southern England.

Understanding the effects of local habitat and wider landscape connectivity factors on tick presence, nymph density and Borrelia species (spp.) prevalence in the tick population is important for identifying the public health risk from Lyme borreliosis. This multi-city study collected data in three southern England cities (Bath, Bristol, and Southampton) during spring, summer, and autumn in 2017. Focusing specifically on urban green space used for recreation which were clearly in urbanised areas, 72 locations were sampled. Additionally, geospatial datasets on urban green space coverage within 250 m and 1 km of sampling points, as well as distance to woodland were incorporated into statistical models. Distance to woodland was negatively associated with tick presence and nymph density, particularly during spring and summer. Furthermore, we observed an interaction effect between habitat and season for tick presence and nymph density, with woodland habitat having greater tick presence and nymph density during spring. Borrelia spp. infected Ixodes ricinus were found in woodland, woodland edge and under canopy habitats in Bath and Southampton. Overall Borrelia spp. prevalence in nymphs was 2.8%, similar to wider UK studies assessing prevalence in Ixodes ricinus in rural areas. Bird-related Borrelia genospecies dominated across sites, suggesting bird reservoir hosts may be important in urban green space settings for feeding and infecting ticks. Whilst overall density of infected nymphs across the three cities was low (0.03 per 100 m2), risk should be further investigated by incorporating data on tick bites acquired in urban settings, and subsequent Lyme borreliosis transmission.

Tahyna virus-A widespread, but neglected mosquito-borne virus in Europe.

Tahyna virus (TAHV) is an orthobunyavirus and was the first arbovirus isolated from mosquitoes in Europe and is associated with floodplain areas as a characteristic biotope, hares as reservoir hosts and the mammal-feeding mosquitoes Aedes vexans as the main vector. The disease caused by TAHV ("Valtice fever") was detected in people with acute flu-like illness in the 1960s, and later the medical significance of TAHV became the subject of many studies. Although TAHV infections are widespread, the prevalence and number of actual cases, clinical manifestations in humans and animals and the ecology of transmission by mosquitoes and their vertebrate hosts are rarely reported. Despite its association with meningitis in humans, TAHV is a neglected human pathogen with unknown public health importance in Central Europe, and a potential emerging disease threat elsewhere in Europe due to extreme summer flooding events.

Detection of mosquito-only flaviviruses in Europe.

The genus Flavivirus, family Flaviviridae, includes a number of important arthropod-transmitted human pathogens such as dengue viruses, West Nile virus, Japanese encephalitis virus and yellow fever virus. In addition, the genus includes flaviviruses without a known vertebrate reservoir, which have been detected only in insects, particularly in mosquitoes, such as cell fusing agent virus, Kamiti River virus, Culex flavivirus, Aedes flavivirus, Quang Binh virus, Nakiwogo virus and Calbertado virus. Reports of the detection of these viruses with no recognized pathogenic role in humans are increasing in mosquitoes collected around the world, particularly in those sampled in entomological surveys targeting pathogenic flaviviruses. The presence of six potential flaviviruses, detected from independent European arbovirus surveys undertaken in the Czech Republic, Italy, Portugal, Spain and the UK between 2007 and 2010, is reported in this work. Whilst the Aedes flaviviruses, detected in Italy from Aedes albopictus mosquitoes, had already been isolated in Japan, the remaining five viruses have not been reported previously: one was detected in Italy, Portugal and Spain from Aedes mosquitoes (particularly from Aedes caspius), one in Portugal and Spain from Culex theileri mosquitoes, one in the Czech Republic and Italy from Aedes vexans, one in the Czech Republic from Aedes vexans and the last in the UK from Aedes cinereus. Phylogenetic analysis confirmed the close relationship of these putative viruses to other insect-only flaviviruses.

Questing Ixodes ricinus ticks and Borrelia spp. in urban green space across Europe: A review.

For more than three decades, it has been recognized that Ixodes ricinus ticks occur in urban green space in Europe and that they harbour multiple pathogens linked to both human and animal diseases. Urban green space use for health and well-being, climate mitigation or biodiversity goals is promoted, often without consideration for the potential impact on tick encounters or tick-borne disease outcomes. This review synthesizes the results of over 100 publications on questing I. ricinus and Borrelia spp. infections in ticks in urban green space in 24 European countries. It presents data on several risk indicators for Lyme borreliosis and highlights key research gaps and recommendations for future studies. Across Europe, mean density of I. ricinus in urban green space was 6.9 (range; 0.1-28.8) per 100 m2 and mean Borrelia prevalence was 17.3% (range; 3.1%-38.1%). Similar density estimates were obtained for nymphs, which had a Borrelia prevalence of 14.2% (range; 0.5%-86.7%). Few studies provided data on both questing nymph density and Borrelia prevalence, but those that did found an average of 1.7 (range; 0-5.6) Borrelia-infected nymphs per 100 m2 of urban green space. Although a wide range of genospecies were reported, Borrelia afzelii was the most common in most parts of Europe, except for England where B. garinii was more common. The emerging pathogen Borrelia miyamotoi was also found in several countries, but with a much lower prevalence (1.5%). Our review highlights that I. ricinus and tick-borne Borrelia pathogens are found in a wide range of urban green space habitats and across several seasons. The impact of human exposure to I. ricinus and subsequent Lyme borreliosis incidence in urban green space has not been quantified. There is also a need to standardize sampling protocols to generate better baseline data for the density of ticks and Borrelia prevalence in urban areas.

Urban woodland habitat is important for tick presence and density in a city in England.

Urban green spaces provide an opportunity for contact between members of the public and ticks infected with pathogens. Understanding tick distribution within these areas and the drivers for increased tick density or Borrelia infection are important from a risk management perspective. This study aimed to generate data on tick presence, nymph density and Borrelia infection across a range of urban green space habitats, in order to identify those that may potentially present a higher risk of Lyme borreliosis to members of the public. Several sites were visited across the English city of Bath during 2015 and 2016. Tick presence was confirmed in all habitats surveyed, with increased likelihood in woodland and woodland edge. Highest nymph densities were also reported in these habitats, along with grassland during one of the sampling years. Adult ticks were more likely to be infected compared to nymphs, and the highest densities of infected nymphs were associated with woodland edge habitat. In addition to Lyme borreliosis causing Borrelia genospecies, Borrelia miyamotoi was also detected at several sites. This study adds to the growing evidence that urban green space habitats present a public health risk from tick bites, and this has implications for many policy areas including health and wellbeing, climate adaptation and urban green space planning.

One Health Approach to Tick and Tick-Borne Disease Surveillance in the United Kingdom.

Where ticks are found, tick-borne diseases can present a threat to human and animal health. The aetiology of many of these important diseases, including Lyme disease, bovine babesiosis, tick-borne fever and louping ill, have been known for decades whilst others have only recently been documented in the United Kingdom (UK). Further threats such as the importation of exotic ticks through human activity or bird migration, combined with changes to either the habitat or climate could increase the risk of tick-borne disease persistence and transmission. Prevention of tick-borne diseases for the human population and animals (both livestock and companion) is dependent on a thorough understanding of where and when pathogen transmission occurs. This information can only be gained through surveillance that seeks to identify where tick populations are distributed, which pathogens are present within those populations, and the periods of the year when ticks are active. To achieve this, a variety of approaches can be applied to enhance knowledge utilising a diverse range of stakeholders (public health professionals and veterinarians through to citizen scientists). Without this information, the application of mitigation strategies to reduce pathogen transmission and impact is compromised and the ability to monitor the effects of climate change or landscape modification on the risk of tick-borne disease is more challenging. However, as with many public and animal health interventions, there needs to be a cost-benefit assessment on the most appropriate intervention applied. This review will assess the challenges of tick-borne diseases in the UK and argue for a cross-disciplinary approach to their surveillance and control.

Prevalence of Anaplasma phagocytophilum in questing Ixodes ricinus nymphs across twenty recreational areas in England and Wales.

Human granulocytic anaplasmosis and tick-borne fever, affecting livestock, are diseases caused by an infection with the bacterium Anaplasma phagocytophilum. Its transmission dynamics between vertebrate hosts and ticks remain largely unknown and the potential impact on public health in the United Kingdom is unclear. This study aimed to assess the distribution and estimate the prevalence of A. phagocytophilum in questing Ixodes ricinus at recreational locations across England and Wales over six years. An additional objective was to investigate possible associations between prevalence, habitat and presence of ruminant hosts. Ixodes ricinus ticks were collected each spring at 20 recreational locations across England and Wales between 2014 and 2019. Nymphs were tested for infection with A. phagocytophilum by detection of bacterial genome in DNA extracts, targeting the msp2 gene locus. Positive samples were further investigated for the presence of different ecotypes based on the GroEL region. Of 3,919 nymphs tested, the mean infection prevalence was 3.6% [95%CI: 3.1-4.3] and ranged from 0 to 20.4%. Northern England had a higher overall prevalence (4.7% [95%CI: 3.4-6.4]) compared to Southern England (1.8% [95%CI: 1.3-2.5]) and the presence of sheep was associated with higher A. phagocytophilum prevalence (8.4% [95%CI: 6.9-10.1] vs 1.2% [95%CI: 0.8-1.7] when absent). There was also a negative correlation with the prevalence of Borrelia burgdorferi s.l. (causing Lyme borreliosis). When investigating the diversity of A. phagocytophilum, ecotype I accounted for 86.8% of samples and ecotype II for 13.2%. Our study presents an overview of A. phagocytophilum prevalence in questing I. ricinus in recreational areas across England and Wales and discusses the potential public and veterinary health relevance.

Evidence for overwintering and autochthonous transmission of Usutu virus to wild birds following its redetection in the United Kingdom.

Usutu virus (USUV) is an emerging zoonotic arbovirus in Europe, where it primarily impacts Eurasian blackbirds (Turdus merula). For mosquito-borne viruses to persist in temperate areas, transovarial transmission in vectors or overwintering in either hosts or diapausing vectors must occur to facilitate autochthonous transmission. We undertook surveillance of hosts and vectors in 2021 to elucidate whether USUV had overwintered in the United Kingdom (UK) following its initial detection there in 2020. From 175 dead bird submissions, we detected 1 case of USUV infection, in a blackbird, from which a full USUV genome was derived. Using a molecular clock analysis, we demonstrate that the 2021 detection shared a most recent common ancestor with the 2020 Greater London, UK, USUV sequence. In addition, we identified USUV-specific neutralizing antibodies in 10 out of 86 serum samples taken from captive birds at the index site, demonstrating in situ cryptic infection and potential sustained transmission. However, from 4966 mosquitoes, we detected no USUV RNA suggesting that prevalence in the vector community was absent or low during sampling. Combined, these results suggest that USUV overwintered in the UK, thus providing empirical evidence for the continued northward expansion of this vector-borne viral disease. Currently, our detection indicates geographically restricted virus persistence. Further detections over time will be required to demonstrate long-term establishment. It remains unclear whether the UK, and by extension other high-latitude regions, can support endemic USUV infection.