Understanding the disease and economic impact of avirulent avian paramyxovirus type 1 (APMV-1) infection in Great Britain.

Newcastle disease (ND) is a notifiable disease affecting chickens and other avian species caused by virulent strains of Avian paramyxovirus type 1 (APMV-1). While outbreaks of ND can have devastating consequences, avirulent strains of APMV-1 generally cause subclinical infections or mild disease. However, viruses can cause different levels of disease in different species and virulence can evolve following cross-species transmission events. This report describes the detection of three cases of avirulent APMV-1 infection in Great Britain (GB). Case 1 emerged from the 'testing to exclude' scheme in chickens in Shropshire while cases 2 and 3 were made directly from notifiable avian disease investigations in chicken broilers in Herefordshire and on premises in Wiltshire containing ducks and mixed species, respectively). Class II/genotype I.1.1 APMV-1 from case 1 shared 99.94% identity to the Queensland V4 strain of APMV-1. Class II/genotype II APMV-1 was detected from case 2 while the class II/genotype I.2 virus from case 3 aligned closely with strains isolated from Anseriformes. Exclusion of ND through rapid detection of avirulent APMV-1 is important where clinical signs caused by avirulent or virulent APMV-1s could be ambiguous. Understanding the diversity of APMV-1s circulating in GB is critical to understanding disease threat from these adaptable viruses.

Happier during lockdown: a descriptive analysis of self-reported wellbeing in 17,000 UK school students during Covid-19 lockdown.

Relatively little research has focused on children and young people (CYP) whose mental health and wellbeing improved during Covid-19 lockdown measures. We aimed to (1) determine the proportion of CYP who self-reported improvement in their mental wellbeing during the first Covid-19 lockdown and (2) describe the characteristics of this group in relation to their peers. We conducted a descriptive analysis of data from the 2020 OxWell Student Survey, a self-report, cross-sectional survey of English CYP. A total of 16,940 CYP primarily aged 8-18 years reported on change in mental wellbeing during lockdown. We characterised these CYP in terms of school, home, relational, and lifestyle factors as well as feelings about returning to school. One-third (33%) of CYP reported improved mental wellbeing during the first UK national lockdown. Compared with peers who reported no change or deterioration, a higher proportion of CYP with improved mental wellbeing reported improved relationships with friends and family, less loneliness and exclusion, reduced bullying, better management of school tasks, and more sleep and exercise during lockdown. In conclusion, a sizeable minority of CYP reported improved mental wellbeing during lockdown. Determining the reasons why these CYP felt they fared better during lockdown and considering how these beneficial experiences can be maintained beyond the pandemic might provide insights into how to promote the future mental health and wellbeing of school-aged CYP. All those working with CYP now have an opportunity to consider whether a systemic shift is needed in order to understand and realise any learnings from experiences during the pandemic.

Incursion of H5N8 high pathogenicity avian influenza virus (HPAIV) into gamebirds in England.

The 2016-17 European outbreak of H5N8 HPAIV (Clade 2.3.4.4b) affected a wider range of avian species than the previous H5N8 outbreak (2014-15), including an incursion of H5N8 HPAIV into gamebirds in England. Natural infection of captive-reared pheasants (Phasianus colchicus) led to variable disease presentation; clinical signs included ruffled feathers, reluctance to move, bright green faeces, and/or sudden mortality. Several birds exhibited neurological signs (nystagmus, torticollis, ataxia). Birds exhibiting even mild clinical signs maintained substantial levels of virus replication and shedding, with preferential shedding via the oropharyngeal route. Gross pathology was consistent with HPAIV, in gallinaceous species but diphtheroid plaques in oropharyngeal mucosa associated with necrotising stomatitis were novel but consistent findings. However, minimal or modest microscopic pathological lesions were detected despite the systemic dissemination of the virus. Serology results indicated differences in the timeframe of exposure for each case (n = 3). This supported epidemiological conclusions confirming that the movement of birds between sites and other standard husbandry practices with limited hygiene involved in pheasant rearing (including several fomite pathways) contributed to virus spread between premises.

Loneliness and self-harm in adolescents during the first national COVID-19 lockdown: results from a survey of 10,000 secondary school pupils in England.

Adolescents' loneliness and self-harm have received considerable attention during the COVID-19 pandemic with concerns that the socioecological changes taking place would contribute to an escalation of both loneliness and self-harm. However, empirical evidence is scant. We estimated the prevalence of loneliness and self-harm in adolescent school pupils and investigated the association of loneliness and change in loneliness during the UK's first lockdown with self-harm during lockdown in a cross-sectional school survey (OxWell) involving 10,460 12-18-year-olds from south England. Loneliness was measured with four items. Self-harm was ascertained through a detailed questionnaire. The prevalence of loneliness and self-harm were estimated applying post-stratification weights to account for differences between the study sample and the target population. The associations between indicators of loneliness and self-harm were examined using mixed effect models. 1,896 of 10,460 adolescents (18.1%) reported feeling lonely 'often' (weighted proportion 16.8%). 3,802/10,460 (36.4%; weighted proportion 35%) felt more lonely since lockdown. Self-harm during lockdown was reported by 787/10,460 adolescents (7.5%; weighted proportion 6.7%). Controlling for confounders, adolescents who reported feeling lonely 'often' [adjusted odds ratio (aOR) 2.8, 95% CI 2.1-3.9, p < 0.0001] or 'sometimes' (aOR 2.2, 95% CI 1.5-3.2, p < 0.0001) were more likely to self-harm during lockdown relative to adolescents who reported 'never' or 'hardly ever' feeling lonely. Exacerbation in loneliness during lockdown was associated with an increase in the odds of self-harm during lockdown. Loneliness, heightened loneliness and self-harm were common during lockdown and closely linked. It is important to support schools in address loneliness and self-harm as part of efforts to improve well-being as the long tail of the pandemic continues to impact on child and adolescent mental health. Understanding how loneliness and self-harm may co-vary could be important for future self-harm reduction strategies in young persons. Supplementary Information: The online version contains supplementary material available at 10.1007/s12144-022-03651-5.

The True Colours Remote Symptom Monitoring System: A Decade of Evolution.

The True Colours remote mood monitoring system was developed over a decade ago by researchers, psychiatrists, and software engineers at the University of Oxford to allow patients to report on a range of symptoms via text messages, Web interfaces, or mobile phone apps. The system has evolved to encompass a wide range of measures, including psychiatric symptoms, quality of life, and medication. Patients are prompted to provide data according to an agreed personal schedule: weekly, daily, or at specific times during the day. The system has been applied across a number of different populations, for the reporting of mood, anxiety, substance use, eating and personality disorders, psychosis, self-harm, and inflammatory bowel disease, and it has shown good compliance. Over the past decade, there have been over 36,000 registered True Colours patients and participants in the United Kingdom, with more than 20 deployments of the system supporting clinical service and research delivery. The system has been adopted for routine clinical care in mental health services, supporting more than 3000 adult patients in secondary care, and 27,263 adolescent patients are currently registered within Oxfordshire and Buckinghamshire. The system has also proven to be an invaluable scientific resource as a platform for research into mood instability and as an electronic outcome measure in randomized controlled trials. This paper aimed to report on the existing applications of the system, setting out lessons learned, and to discuss the implications for tailored symptom monitoring, as well as the barriers to implementation at a larger scale.

Identification and characterization of a novel tick-borne flavivirus subtype in goats (Capra hircus) in Spain.

In 2011, a neurological disease was reported in a herd of goats (Capra hircus) in Asturias, Spain. Initial sequencing identified the causative agent as louping ill virus (LIV). Subsequently, with the application of whole genome sequencing and phylogenetic analysis, empirical data demonstrates that the LIV-like virus detected is significantly divergent from LIV and Spanish sheep encephalitis virus (SSEV). This virus encoded an amino acid sequence motif at the site of a previously identified marker for differentiating tick-borne flaviviruses that was shared with a virus previously isolated in Ireland in 1968. The significance of these observations reflects the diversity of tick-borne flaviviruses in Europe. These data also contribute to our knowledge of the evolution of tick-borne flaviviruses and could reflect the movement of viruses throughout Europe. Based on these observations, the proposed name for this virus is Spanish goat encephalitis virus (SGEV), to distinguish it from SSEV.

Defining the chemokine basis for leukocyte recruitment during viral encephalitis.

UNLABELLED: The encephalitic response to viral infection requires local chemokine production and the ensuing recruitment of immune and inflammatory leukocytes. Accordingly, chemokine receptors present themselves as plausible therapeutic targets for drugs aimed at limiting encephalitic responses. However, it remains unclear which chemokines are central to this process and whether leukocyte recruitment is important for limiting viral proliferation and survival in the brain or whether it is predominantly a driver of coincident inflammatory pathogenesis. Here we examine chemokine expression and leukocyte recruitment in the context of avirulent and virulent Semliki Forest virus (SFV) as well as West Nile virus infection and demonstrate rapid and robust expression of a variety of inflammatory CC and CXC chemokines in all models. On this basis, we define a chemokine axis involved in leukocyte recruitment to the encephalitic brain during SFV infection. CXCR3 is the most active; CCR2 is also active but less so, and CCR5 plays only a modest role in leukocyte recruitment. Importantly, inhibition of each of these receptors individually and the resulting suppression of leukocyte recruitment to the infected brain have no effect on viral titer or survival following infection with a virulent SFV strain. In contrast, simultaneous blockade of CXCR3 and CCR2 results in significantly reduced mortality in response to virulent SFV infection. In summary, therefore, our data provide an unprecedented level of insight into chemokine orchestration of leukocyte recruitment in viral encephalitis. Our data also highlight CXCR3 and CCR2 as possible therapeutic targets for limiting inflammatory damage in response to viral infection of the brain. IMPORTANCE: Brain inflammation (encephalitis) in response to viral infection can lead to severe illness and even death. This therefore represents an important clinical problem and one that requires the development of new therapeutic approaches. Central to the pathogenesis of encephalitis is the recruitment of inflammatory leukocytes to the infected brain, a process driven by members of the chemokine family. Here we provide an in-depth analysis of the chemokines involved in leukocyte recruitment to the virally infected brain and demonstrate that simultaneous blockade of two of these receptors, namely, CXCR3 and CCR2, does not alter viral titers within the brain but markedly reduces inflammatory leukocyte recruitment and enhances survival in a murine model of lethal viral encephalitis. Our results therefore highlight chemokine receptors as plausible therapeutic targets in treating viral encephalitis.

Sequences Related to Chimay Rhabdovirus Are Widely Distributed in Ixodes ricinus Ticks across England and Wales.

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

Temporal dynamics of interference in Simon and Eriksen tasks considered within the context of a dual-process model.

Behavioral and brain potential measures were employed to compare interference in Eriksen and Simon tasks. Assuming a dual-process model of interference elicited in speeded response tasks, we hypothesized that only lateralized stimuli in the Simon task induce fast S-R priming via direct unconditional processes, while Eriksen interference effects are induced later via indirect conditional processes. Delays to responses for incongruent trials were indeed larger in the Eriksen than in the Simon task. Only lateralized stimuli in the Simon task elicited early S-R priming, maximal at parietal areas. Incongruent flankers in the Eriksen task elicited interference later, visible as a lateralized N2. Eriksen interference also elicited an additional component (N350), which accounted for the larger behavioral interference effects in the Eriksen task. The findings suggest that interference and its resolution involve different processes for Simon and Eriksen tasks.

Missing the context: The challenge of social inequalities to school-based mental health interventions.

Given well-established links between socio-economic adversity and mental health, it is unsurprising that young people's mental health is deteriorating amidst economic crises. The World Health Organisation (WHO) recognises mental health as "crucial to personal, community, and socio-economic development" and outlines goals to reshape environments such as schools to protect mental health. Schools offer an ideal setting to promote wellbeing and prevent mental ill-health during a key developmental window. We describe how social inequalities present a challenge to designing school-based interventions for prevention and promotion for mental health and wellbeing, and suggest priorities to aid and evaluate their effectiveness.

Young People's Mental Health Changes, Risk, and Resilience During the COVID-19 Pandemic.

Importance: As young people's mental health difficulties increase, understanding risk and resilience factors under challenging circumstances becomes critical. Objective: To explore the outcomes of the COVID-19 pandemic on secondary school students' mental health difficulties, as well as the associations with individual, family, friendship, and school characteristics. Design, Setting, and Participants: For this cohort study, follow-up data from the My Resilience in Adolescence (MYRIAD) cluster randomized clinical trial were collected across 2 representative UK cohorts. Mainstream UK secondary schools with a strategy and structure to deliver social-emotional learning, with an appointed head teacher, and that were not rated "inadequate" in their latest official inspection were recruited. A total of 5663 schools were approached, 532 showed interest, and 84 consented. Cohort 1 included 12 schools and 864 students, and cohort 2 included 72 schools and 6386 students. COVID-19 was declared a pandemic after cohort 1 had completed all assessments (September 2018 to January 2020), but cohort 2 had not (September 2019 to June 2021). Exposures: Cohort 2 was exposed to the COVID-19 pandemic, including 3 national lockdowns. Associations of individual, family, friendship, and school characteristics with students' mental health were explored. Main Outcomes and Measures: Changes in students' risk for depression (Center for Epidemiological Studies-Depression scale); social, emotional, and behavioral difficulties (Strengths and Difficulties Questionnaire); and mental well-being (Warwick-Edinburgh Mental Well-Being Scale). Results: Of the 7250 participants included, the mean (SD) age was 13.7 (0.6) years, 3947 (55.4%) identified as female, and 5378 (73.1%) self-reported their race as White. Twelve schools and 769 of the 864 students (89.0%) in cohort 1 and 54 schools and 2958 of the 6386 students (46.3%) in cohort 2 provided data and were analyzed. Mental health difficulties increased in both cohorts but to a greater extent among students exposed to the pandemic, including for risk of depression (adjusted mean difference [AMD], 1.91; 95% CI, 1.07-2.76); social, emotional, and behavioral difficulties (AMD, 0.76; 95% CI, 0.33-1.18); and mental well-being (AMD, -2.08; 95% CI, -2.80 to -1.36). Positive school climate, high home connectedness, and having a friend during lockdown were protective factors during the pandemic. Female gender and initial low risk for mental health difficulties were associated with greater mental health deteriorations. Partial school attendance during lockdown was associated with better adjustment than no attendance when returning to school. Conclusions and Relevance: This cohort study of secondary school students demonstrated that to promote mental health and adjustment, policy interventions should foster home connectedness, peer friendship, and school climate; avoid full school closures; and consider individual differences.

Rapid molecular detection methods for arboviruses of livestock of importance to northern Europe.

Arthropod-borne viruses (arboviruses) have been responsible for some of the most explosive epidemics of emerging infectious diseases over the past decade. Their impact on both human and livestock populations has been dramatic. The early detection either through surveillance or diagnosis of virus will be a critical feature in responding and resolving the emergence of such epidemics in the future. Although some of the most important emerging arboviruses are human pathogens, this paper aims to highlight those diseases that primarily affect livestock, although many are zoonotic and some occasionally cause human mortality. This paper also highlights the molecular detection methods specific to each virus and identifies those emerging diseases for which a rapid detection methods are not yet developed.

JMM Profile: Japanese encephalitis virus: an emerging threat.

Japanese encephalitis (JE) is an infection that occurs predominantly in Asia and the Pacific Islands. It is transmitted by mosquito bites, with the main vector being Culex tritaeniorhynchus, and is maintained in enzootic cycles involving pigs, wild birds and mosquitoes. JE is caused by infection with Japanese encephalitis virus (JEV), a zoonotic pathogen that also causes disease in mammals such as pigs and horses. In humans, most symptoms are mild or flu-like but can progress to encephalitis. Pigs are considered amplification hosts, and sows may have gestational complications. Horses may exhibit neurological signs. Detection of the virus can be confirmed by serological or molecular laboratory tests. Vaccination offers protection against JEV infection in humans, pigs and horses. Whilst there is no effective treatment of JE, human cases may require hospitalization for supportive therapy, which may include administration of fluids, oxygen and medication to treat symptoms.

Utilization and Acceptability of Formal and Informal Support for Adolescents Following Self-Harm Before and During the First COVID-19 Lockdown: Results From a Large-Scale English Schools Survey.

Background: Little is known about the perceived acceptability and usefulness of supports that adolescents have accessed following self-harm, especially since the onset of the COVID-19 pandemic. Aims: To examine the utilization and acceptability of formal, informal, and online support accessed by adolescents following self-harm before and during the pandemic. Method: Cross-sectional survey (OxWell) of 10,560 secondary school students aged 12-18 years in the south of England. Information on self-harm, support(s) accessed after self-harm, and satisfaction with support received were obtained via a structured, self-report questionnaire. No tests for significance were conducted. Results: 1,457 (12.5%) students reported having ever self-harmed and 789 (6.7%) reported self-harming during the first national lockdown. Informal sources of support were accessed by the greatest proportion of respondents (friends: 35.9%; parents: 25.0%). Formal sources of support were accessed by considerably fewer respondents (Child and Adolescent Mental Health Services: 12.1%; psychologist/ psychiatrist: 10.2%; general practitioner: 7.4%). Online support was accessed by 8.6% of respondents, and 38.3% reported accessing no support at all. Informal sources of support were rated as most helpful, followed by formal sources, and online support. Of the respondents who sought no support, 11.3% reported this as being helpful. Conclusions: More than a third of secondary school students in this sample did not seek any help following self-harm. The majority of those not seeking help did not find this to be a helpful way of coping. Further work needs to determine effective ways of overcoming barriers to help-seeking among adolescents who self-harm and improving perceived helpfulness of the supports accessed.

Sleep in the time of COVID-19: findings from 17000 school-aged children and adolescents in the UK during the first national lockdown.

STUDY OBJECTIVES: Sleep is essential to young people's wellbeing, yet may be constricted by the adolescent delayed sleep phase coupled with school start times. COVID-19 restrictions caused major disruptions to everyday routines, including partial school closures. We set out to understand changes in students' self-reported sleep quality, and associations with mental wellbeing and interpersonal functioning, during these restrictions. METHODS: The OxWell school survey-a cross-sectional online survey-collected data from 18 642 children and adolescents (aged 8-19 years, 60% female, school year 4-13) from 230 schools in southern England, in June-July 2020. Participants completed self-report measures of the impact of COVID-19 restrictions on sleep quality, happiness, and social relationships. Sleep timing was compared with data collected from 4222 young people in 2019. RESULTS: Females and older adolescents were more likely to report deteriorations in sleep during the national lockdown. Regression analysis revealed that changes in happiness (ß = .34) and how well students were getting on with others in their household (ß = .07) predicted change in sleep quality. Students' bedtimes and wake times were later, and sleep duration was longer in 2020 compared to the 2019 survey. Secondary school students reported the greatest differences, especially later wake times. CONCLUSIONS: During COVID-19 restrictions, sleep patterns consistent with adolescent delayed sleep phase were observed, with longer sleep times for secondary school students in particular. Perceived deteriorations in sleep quality were associated with reductions in happiness and interpersonal functioning, highlighting the importance of including sleep measures in adolescent wellbeing research.

Batai Orthobunyavirus: An Emerging Mosquito-Borne Virus in Europe.

Batai virus (BATV) is a zoonotic orthobunyavirus transmitted by a wide range of mosquito vectors. The virus is distributed throughout Asia and parts of Africa and has been sporadically detected in several European countries. There is increasing evidence that BATV is emerging in Europe as a potential threat to both animal and human health, having been detected in mosquitoes, mammals, birds and humans. In recent years, serological surveillance in cattle, sheep and goats has suggested an antibody prevalence of up to 46% in European livestock, although human serological prevalence remains generally low. However, the recent and continued spread of invasive mosquito species into Europe may facilitate the establishment of competent populations of mosquitoes leading to increased BATV transmission. Migratory birds may also potentially facilitate the emergence of BATV in geographical locations where it was previously undetected. Although BATV has the potential to cause disease in humans and livestock, our understanding of the impact in wild animal populations is extremely limited. Therefore, there is a need for increased surveillance for BATV in mosquitoes, livestock, wild mammals and birds in Europe to understand the true impact of this virus.

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.

Blinding efficacy and adverse events following repeated transcranial alternating current, direct current, and random noise stimulation.

As transcranial electrical stimulation (tES) protocols advance, assumptions underlying the technique need to be retested to ensure they still hold. Whilst the safety of stimulation has been demonstrated mainly for a small number of sessions, and small sample size, adverse events (AEs) following multiple sessions remain largely untested. Similarly, whilst blinding procedures are typically assumed to be effective, the effect of multiple stimulation sessions on the efficacy of blinding procedures also remains under question. This is especially relevant in multisite projects where small unintentional variations in protocol could lead to inter-site difference. We report AE and blinding data from 1,019 participants who received up to 11 semi-consecutive sessions of active or sham transcranial alternating current stimulation (tACS), direct current stimulation (tDCS), and random noise stimulation (tRNS), at 4 sites in the UK and US. We found that AEs were often best predicted by factors other than tES, such as testing site or session number. Results from the blinding analysis suggested that blinding was less effective for tDCS and tACS than tRNS. The occurrence of AEs did not appear to be linked to tES despite the use of smaller electrodes or repeated delivery. However, blinding efficacy was impacted in tES conditions with higher cutaneous sensation, highlighting a need for alternative stimulation blinding protocols. This may be increasingly necessary in studies wishing to deliver stimulation with higher intensities.

Susceptibility of Aedes albopictus and Culex quinquefasciatus to Japanese encephalitis virus.

BACKGROUND: Japanese encephalitis virus (JEV) is the principal cause of mosquito-borne encephalitis in human populations within Asia. If introduced into new geographic areas, it could have further implications for public and animal health. However, potential mosquito vectors for virus transmission have not been fully investigated. The Asian tiger mosquito, Aedes albopictus, has emerged in Europe and is now expanding its geographical range into more northerly latitudes. Culex quinquefasciatus, although absent from Europe, has been detected in Turkey, a country with territory in Europe, and could act as a vector for JEV in other regions. To assess the risk of these invasive species acting as vectors for JEV and therefore potentially contributing to its geographical expansion, we have investigated the vector competence of Ae. albopictus and Cx. quinquefasciatus. METHODS: Two colonised lines of Ae. albopictus (Italy and Spain) and a line of Cx. quinquefasciatus (Tanzania) were compared for susceptibility to infection by oral feeding with JEV strain SA-14, genotype III at 106 PFU/ml and maintained at 25 °C. Specimens were processed at 7 and 14 days post-inoculation (dpi). Rates of infection, dissemination and transmission were assessed through detection of viral RNA by real-time polymerase chain reaction (RT-PCR) in mosquito body, legs and saliva, respectively, at each time point. Where possible, infection and dissemination were confirmed by immunohistochemical (IHC) detection of the JEV envelope protein. RESULTS: Aedes albopictus from Italy showed no susceptibility to infection with JEV strain SA-14. Conversely, Ae. albopictus colonised in Spain was susceptible and 100% of infected mosquitoes that were subjected to saliva screening expressed viral RNA at 14 dpi. Culex quinquefasciatus was highly susceptible to infection as early as 7 dpi and 50% of infected mosquitoes that were subjected to saliva screening expressed viral RNA at 14 dpi. Infection and dissemination were confirmed in Cx. quinquefasciatus by IHC detection of JEV envelope protein in both the mid-gut and salivary glands. CONCLUSIONS: Aedes albopictus from two different locations in Europe range from being susceptible to JEV and capable of transmission through to being resistant. Culex quinquefasciatus also appears highly susceptible; therefore, both species could potentially act as vectors for JEV and facilitate the emergence of JEV into new regions.