An integrated public health response to an outbreak of Murray Valley encephalitis virus infection during the 2022-2023 mosquito season in Victoria.

Introduction: Murray Valley encephalitis virus (MVEV) is a mosquito-borne flavivirus known to cause infrequent yet substantial human outbreaks around the Murray Valley region of south-eastern Australia, resulting in significant mortality. Methods: The public health response to MVEV in Victoria in 2022-2023 included a climate informed pre-season risk assessment, and vector surveillance with mosquito trapping and laboratory testing for MVEV. Human cases were investigated to collect enhanced surveillance data, and human clinical samples were subject to serological and molecular testing algorithms to assess for co-circulating flaviviruses. Equine surveillance was carried out via enhanced investigation of cases of encephalitic illness. Integrated mosquito management and active health promotion were implemented throughout the season and in response to surveillance signals. Findings: Mosquito surveillance included a total of 3,186 individual trapping events between 1 July 2022 and 20 June 2023. MVEV was detected in mosquitoes on 48 occasions. From 2 January 2023 to 23 April 2023, 580 samples (sera and CSF) were tested for flaviviruses. Human surveillance detected 6 confirmed cases of MVEV infection and 2 cases of "flavivirus-unspecified." From 1 September 2022 to 30 May 2023, 88 horses with clinical signs consistent with flavivirus infection were tested, finding one probable and no confirmed cases of MVE. Discussion: The expanded, climate-informed vector surveillance system in Victoria detected MVEV in mosquitoes in advance of human cases, acting as an effective early warning system. This informed a one-health oriented public health response including enhanced human, vector and animal surveillance, integrated mosquito management, and health promotion.

Serologic evidence of West Nile virus and Saint Louis encephalitis virus in horses from Southern Brazil.

Flaviviruses as West Nile virus (WNV), Saint Louis encephalitis virus (SLEV), Ilhéus virus (ILHV), and Rocio virus (ROCV) are previously reported in different Brazilian regions, but studies in Southern Brazil are still scarce. To improve the information regarding flaviviruses in Southern Brazil, horse serum samples were analyzed using RT-qPCR and a commercial ELISA-Ab against WNV followed by PRNT75. All 1000 samples analyzed by real-time RT-PCR resulted negative. The 465 subsampled samples were analyzed by a commercial ELISA-Ab against WNV, and the 18.5% (86/465) positive samples were further analyzed by PRNT75. In the PRNT75, 13/86 and 2/86 horses were positive for SLEV and WNV, respectively. It was observed that 5.8% (13/226) of the farms presented at least one positive animal for SLEV in PRNT75, whereas 0.9% (2/226) for WNV. Apart from the lower seroprevalences identified when compared to data previously reported in other Brazilian regions, our results suggest that public health professionals must be aware of the presence of these potential zoonotic pathogens.

Mosquito-Borne Viruses and Non-Human Vertebrates in Australia: A Review.

Mosquito-borne viruses are well recognized as a global public health burden amongst humans, but the effects on non-human vertebrates is rarely reported. Australia, houses a number of endemic mosquito-borne viruses, such as Ross River virus, Barmah Forest virus, and Murray Valley encephalitis virus. In this review, we synthesize the current state of mosquito-borne viruses impacting non-human vertebrates in Australia, including diseases that could be introduced due to local mosquito distribution. Given the unique island biogeography of Australia and the endemism of vertebrate species (including macropods and monotremes), Australia is highly susceptible to foreign mosquito species becoming established, and mosquito-borne viruses becoming endemic alongside novel reservoirs. For each virus, we summarize the known geographic distribution, mosquito vectors, vertebrate hosts, clinical signs and treatments, and highlight the importance of including non-human vertebrates in the assessment of future disease outbreaks. The mosquito-borne viruses discussed can impact wildlife, livestock, and companion animals, causing significant changes to Australian ecology and economy. The complex nature of mosquito-borne disease, and challenges in assessing the impacts to non-human vertebrate species, makes this an important topic to periodically review.

Encephalitic Arboviruses of Africa: Emergence, Clinical Presentation and Neuropathogenesis.

Many mosquito-borne viruses (arboviruses) are endemic in Africa, contributing to systemic and neurological infections in various geographical locations on the continent. While most arboviral infections do not lead to neuroinvasive diseases of the central nervous system, neurologic diseases caused by arboviruses include flaccid paralysis, meningitis, encephalitis, myelitis, encephalomyelitis, neuritis, and post-infectious autoimmune or memory disorders. Here we review endemic members of the Flaviviridae and Togaviridae families that cause neurologic infections, their neuropathogenesis and host neuroimmunological responses in Africa. We also discuss the potential for neuroimmune responses to aide in the development of new diagnostics and therapeutics, and current knowledge gaps to be addressed by arbovirus research.

[Epidemiological and spatial-temporal distribution of several natural focus diseases in Gansu province, 2014-2018].

Objective: To analyze the epidemiological and spatial-temporal distribution of Brucellosis, epidemic encephalitis B and hemorrhagic fever with renal syndrome (HFRS) in Gansu province during 2014-2018 so as to provide evidence for the prevention and control of those diseases. Methods: A database was established in Gansu province from 2014 to 2018, using the geographical information system. A spatial distribution map was drawn, with trend analysis and space-time clustering used to study the 3-dimention of the diseases, by using both ArcGIS 10.5 and SaTScan 9.6 softwares. Results: Results from the trend surface analysis showed that the incidence of Brucellosis decreased gradually from north to south parts while the U type curve could reflect the distribution from the east to the west areas. Incidence of epidemic encephalitis B decreased significantly from south to north areas in the province, with incidence higher in the eastern than in the mid-west region. Difference on the incidence of HFRS was not significantly visible in the eastern and western regions, while the incidence was slightly higher in the southern than the northern parts of the province. Spatial and space-time clustering did exist among the 3 diseases in Gansu from 2014 to 2018. The areas with clusters of Brucellosis appeared in the eastern parts during 2014-2015, including 19 counties. The areas with secondary clusters of Brucellosis were seen in the Hexi district, including 4 counties, during 2017-2018. The areas with high incidence of epidemic encephalitis B were clustered in the middle and southeast areas, including 32 counties, during 2017-2018. Areas with most clusters of HFRS appeared in Min county of Dingxi city in 2018, with the areas of secondary clusters in 8 counties of the eastern areas in 2018. Conclusions: The overall incidence rates of the 3 natural focus diseases were in a upward trend and showing obvious characteristics on spatial clustering. According to the distributive characteristics, effective measures should be developed accordingly.

Ongoing and emerging arbovirus threats in Europe.

During the last decades, arboviruses that are endemic in Europe have expanded their geographic range and caused an increasing number of human outbreaks. These viruses include West Nile virus, which is expanding its area of circulation in central and southern Europe; Usutu virus, with increasing evidence of a role in human disease; tick-borne encephalitis virus, which is being detected in northern areas and at higher altitudes as a consequence of climate warming; Crimean-Congo hemorrhagic fever virus, which is endemic in Eastern Europe and the Middle East, but has been recently detected in Spain; other viruses, such as California encephalitis virus antigenic group, which circulate in northern and central Europe but whose relevance for human disease in largely unknown. In addition, the rise in global travel and trade has posed Europe to an increased risk of introduction and expansion of exotic arthropod vectors and autochthonous transmission of arboviruses, like dengue and chikungunya viruses, following new introductions from endemic areas. Implementation of integrated arbovirus surveillance programs has been crucial to adopt proper control measures. The identification of emerging outbreaks is however challenging and requires a high degree of awareness and laboratory capacity, especially for the most neglected but potentially threatening pathogens.

The ecology and epidemiology of Ross River and Murray Valley encephalitis viruses in Western Australia: examples of One Health in Action.

Arboviruses are maintained and transmitted through an alternating biological cycle in arthropods and vertebrates, with largely incidental disease in humans and animals. As such, they provide excellent examples of One Health, as their health impact is inextricably linked to their vertebrate hosts, their arthropod vectors and the environment. Prevention and control requires a comprehensive understanding of these interactions, and how they may be effectively and safely modified. This review concentrates on human disease due to Ross River and Murray Valley encephalitis viruses, the two major arboviral pathogens in Australia. It describes how their pattern of infection and disease is influenced by natural climatic and weather patterns, and by anthropogenic activities. The latter includes human-mediated environmental manipulations, such as water impoundment infrastructures, human movements and migration, and community and social changes, such as urban spread into mosquito larval habitats. Effective interventions need to be directed at the environmental precursors of risk. This can best be achieved using One Health approaches to improve collaboration and coordination between different disciplines and cross-sectoral jurisdictions in order to develop more holistic mitigation and control procedures, and to address poorly understood ecological issues through multidisciplinary research.

Evidence of exposure of laughing doves (Spilopelia senegalensis) to West Nile and Usutu viruses in southern Tunisian oases.

It has previously been suggested that southern Tunisian oases may be suitable areas for the circulation of flaviviruses. In order to anticipate and prevent possible epidemiological spread of flaviviruses in humans and domestic animals, the ecology of their transmission in the oasis system needs to be better understood. Thus, the aim of this study was to assess the seroprevalence of anti-flavivirus antibodies in the laughing dove (Spilopelia senegalensis), an abundant resident bird in Tunisian oases. Anti-flavivirus antibodies were detected in 17% of sampled doves. Ten per cent of the total tested doves were West Nile virus (WNV) seropositive and 4% were Usutu virus (USUV) seropositive, which provides the first evidence of USUV circulation in Tunisian birds. We also found that the occurrence probability of anti-flavivirus antibodies in dove plasma increased with decreasing distance to coast, suggesting that doves inhabiting coastal oases were more exposed to flaviviruses compared with those inhabiting inland oases. We also found significantly higher antibody occurrence probability in adult doves compared with young doves, which underlines the effect of exposure time. Overall, our results suggest that the laughing dove may be used for WNV and USUV surveillance in southern Tunisia. They also stress the need for investigations combining data on birds and mosquitoes to better understand the ecological factors governing the circulation of flaviviruses in this area.

Fatal Infection with Murray Valley Encephalitis Virus Imported from Australia to Canada, 2011.

Murray Valley encephalitis virus (MVEV), a flavivirus belonging to the Japanese encephalitis serogroup, can cause severe clinical manifestations in humans. We report a fatal case of MVEV infection in a young woman who returned from Australia to Canada. The differential diagnosis for travel-associated encephalitis should include MVEV, particularly during outbreak years.

Climate change, vector-borne diseases and working population.

INTRODUCTION: Risks associated with climate change are increasing worldwide and the global effects include altered weather and precipitation patterns, rising temperatures and others; human health can be affected directly and indirectly. This paper is an overview of literature regarding climate changes, their interaction with vector-borne diseases and impact on working population. MATERIALS AND METHODS: Articles regarding climate changes as drivers of vector-borne diseases and evidences of occupational cases have been picked up by public databank. Technical documents were also included in the study. RESULTS: Evidences regarding the impact of climate changes on vector-borne diseases in Europe, provided by the analysis of the literature, are presented. DISCUSSION: Climate-sensitive vector-borne diseases are likely to be emerging due to climate modifications, with impacts on public and occupational health. However, other environmental and anthropogenic drivers such as increasing travelling and trade, deforestation and reforestation, altered land use and urbanization can influence their spread. Further studies are necessary to better understand the phenomenon and implementation of adaptation strategies to protect human health should be accelerated and strengthened.

La Crosse Encephalitis: A Persistent Arboviral Threat in North Carolina.

Mosquito-borne diseases remain a significant cause of economic, social, and health burdens in North Carolina. Although recently overshadowed by emerging threats such as chikungunya virus and Zika virus, La Crosse virus and other endemic arboviruses remain persistent environmental health hazards. Indeed, La Crosse virus, West Nile virus, and Eastern equine encephalitis virus accounted for more than 98% of the reported human arboviral diseases acquired in North Carolina in the past decade. Arbovirus infection is increasingly prevalent in Western North Carolina, with La Crosse encephalitis being endemic in this area. While infections are often asymptomatic and seldom fatal, the long-term neurologic sequelae of La Crosse encephalitis represent a significant burden.

Use of Testing for West Nile Virus and Other Arboviruses.

In the United States, the most commonly diagnosed arboviral disease is West Nile virus (WNV) infection. Diagnosis is made by detecting WNV IgG or viral genomic sequences in serum or cerebrospinal fluid. To determine frequency of this testing in WNV-endemic areas, we examined the proportion of tests ordered for patients with meningitis and encephalitis at 9 hospitals in Houston, Texas, USA. We identified 751 patients (567 adults, 184 children), among whom 390 (52%) experienced illness onset during WNV season (June-October). WNV testing was ordered for 281 (37%) of the 751; results indicated acute infection for 32 (11%). Characteristics associated with WNV testing were acute focal neurologic deficits; older age; magnetic resonance imaging; empirically prescribed antiviral therapy; worse clinical outcomes: and concomitant testing for mycobacterial, fungal, or other viral infections. Testing for WNV is underutilized, and testing of patients with more severe disease raises the possibility of diagnostic bias in epidemiologic studies.

Seroprevalence of West Nile and Usutu viruses in military working horses and dogs, Morocco, 2012: dog as an alternative WNV sentinel species?

A serosurvey of 349 military working horses and 231 military working dogs was conducted in ten sites in Morocco in 2012. This survey revealed a high level of exposure of these animals to flaviviruses: seroprevalence rates of 60% in horses and of 62% in dogs were observed using a competitive West Nile virus (WNV) enzyme-linked immunosorbent assay (cELISA). Seroneutralization test results showed that the majority of cELISA-positive results were due to exposure to WNV. Further assays conducted in vaccinated horses with a DIVA (Differentiating Infected from Vaccinated Animals) test indicated that anti-WNV antibodies had been stimulated through WNV natural infection. Moreover, in both species, seroneutralization tests suggested an exposure to Usutu virus (USUV). Data analysis did not show any significant difference of cELISA seropositivity risk between horses and dogs. Dogs may thus represent an interesting alternative to equines for the serological surveillance of WNV or USUV circulation, especially in areas where equine vaccination precludes passive surveillance (based on the detection of West Nile fever cases) in horses.

A Bayesian Belief Network for Murray Valley encephalitis virus risk assessment in Western Australia.

BACKGROUND: Murray Valley encephalitis virus (MVEV) is a clinically important virus in Australia responsible for a number of epidemics over the past century. Since there is no vaccine for MVEV, other preventive health measures to curtail its spread must be considered, including the development of predictive risk models and maps to help direct public health interventions. This article aims to support these approaches by presenting a model for assessing MVEV risk in Western Australia (WA). METHODS: A Bayesian Belief Network (BBN) for assessing MVEV risk was developed and used to quantify and map disease risks in WA. The model combined various abiotic, biotic, and anthropogenic factors that might affect the risk of MVEV into a predictive framework, based on the ecology of the major mosquito vector and waterbird hosts of MVEV. It was further refined and tested using retrospective climate data from 4 years (2000, 2003, 2009, and 2011). RESULTS: Implementing the model across WA demonstrated that it could predict locations of human MVEV infection and sentinel animal seroconversion in the 4 years tested with some degree of accuracy. In general, risks are highest in the State's north and lower in the south. The model predicted that short-term climate change, based on the Intergovernmental Panel on Climate Change's A1B emissions scenario, would decrease MVEV risks in summer and autumn, largely due to higher temperatures decreasing vector survival. CONCLUSIONS: To our knowledge, this is the first model to use a BBN to quantify MVEV risks in WA. The models and maps developed here may assist public health agencies in preparing for and managing Murray Valley encephalitis in the future. In its current form, the model is knowledge-driven and based on the analysis of potential risk factors that affect the dynamics of MVEV using retrospective data. Further work and additional testing should be carried out to test its validity in future years.

Long-term outcomes of Murray Valley encephalitis cases in Western Australia: what have we learnt?

BACKGROUND: Murray Valley encephalitis virus (MVEV) is a mosquito-borne flavivirus that causes encephalitis in some cases of infection. It is endemic in Northern Australia and cases occasionally occur in South Eastern Australia. The long-term sequelae of MVEV infection have not previously been well described. AIM: To investigate the long-term sequelae of MVEV infection. METHODS: This was a descriptive case series of all clinical MVEV infections using data linkage and standard surveys. Hospital admissions, emergency department, psychiatric outpatients and mortality data were obtained. We attempted to follow-up all 53 cases of MVEV clinical infection that occurred in Western Australia from 1978 to 2011 inclusive. Two cases opted out of the study. RESULTS: We followed-up 39 surviving cases. Seven of the nine with paralysis or paresis were under 5 years and they fared worse than other patients, requiring lengthy hospitalisation (median duration 133 days). Two died due to complications of quadriplegia following a total of 691 days in hospital. Nine surviving patients, including two with non-encephalitic illness, required care for depression and other psychiatric conditions following MVEV infection. Two patients who were discharged with neurological sequelae had no further documented hospital occasions of service but reported ongoing challenges with cognitive dysfunction and inability to work. CONCLUSIONS: This is the first study of long-term outcomes of Murray Valley encephalitis that included cases with no obvious sequelae at discharge. In spite of the small numbers involved, the study demonstrated the significant medical and social burden due to MVEV in Australia.

The Molecular Epidemiology and Evolution of Murray Valley Encephalitis Virus: Recent Emergence of Distinct Sub-lineages of the Dominant Genotype 1.

BACKGROUND: Recent increased activity of the mosquito-borne Murray Valley encephalitis virus (MVEV) in Australia has renewed concerns regarding its potential to spread and cause disease. METHODOLOGY/PRINCIPAL FINDINGS: To better understand the genetic relationships between earlier and more recent circulating strains, patterns of virus movement, as well as the molecular basis of MVEV evolution, complete pre-membrane (prM) and Envelope (Env) genes were sequenced from sixty-six MVEV strains from different regions of the Australasian region, isolated over a sixty year period (1951-2011). Phylogenetic analyses indicated that, of the four recognized genotypes, only G1 and G2 are contemporary. G1 viruses were dominant over the sampling period and found across the known geographic range of MVEV. Two distinct sub-lineages of G1 were observed (1A and 1B). Although G1B strains have been isolated from across mainland Australia, Australian G1A strains have not been detected outside northwest Australia. Similarly, G2 is comprised of only Western Australian isolates from mosquitoes, suggesting G1B and G2 viruses have geographic or ecological restrictions. No evidence of recombination was found and a single amino acid substitution in the Env protein (S332G) was found to be under positive selection, while several others were found to be under directional evolution. Evolutionary analyses indicated that extant genotypes of MVEV began to diverge from a common ancestor approximately 200 years ago. G2 was the first genotype to diverge, followed by G3 and G4, and finally G1, from which subtypes G1A and G1B diverged between 1964 and 1994. CONCLUSIONS/SIGNIFICANCE: The results of this study provides new insights into the genetic diversity and evolution of MVEV. The demonstration of co-circulation of all contemporary genetic lineages of MVEV in northwestern Australia, supports the contention that this region is the enzootic focus for this virus.

Co-circulation of Usutu virus and West Nile virus in a reed bed ecosystem.

BACKGROUND: Mosquito-borne flaviviruses are a major public health threat in many countries worldwide. In Central Europe, West Nile virus (WNV) and Usutu virus (USUV), both belonging to the Japanese encephalitis virus group (Flaviviridae) have emerged in the last decennium. Surveillance of mosquito vectors for arboviruses is a sensitive tool to evaluate virus circulation and consequently to estimate the public health risk. METHODS: Mosquitoes (Culicidae) were collected at South-Moravian (Czech Republic) fishponds between 2010 and 2014. A total of 61,770 female Culex modestus Ficalbi mosquitoes, pooled to 1,243 samples, were examined for flaviviruses by RT-PCR. RESULTS: One pool proved positive for USUV RNA. Phylogenetic analysis demonstrated that this Czech USUV strain is closely related to Austrian and other Central European strains of the virus. In addition, nine strains of WNV lineage 2 were detected in Cx. modestus collected in the same reed bed ecosystem. CONCLUSIONS: This is the first detection of USUV in Cx. modestus. The results indicate that USUV and WNV may co-circulate in a sylvatic cycle in the same habitat, characterised by the presence of water birds and Cx. modestus mosquitoes, serving as hosts and vectors, respectively, for both viruses.

A review of West Nile and Usutu virus co-circulation in Europe: how much do transmission cycles overlap?

Due to the increasing global spread of arboviruses, the geographic extent of virus co-circulation is expanding. This complicates the diagnosis of febrile conditions and can have direct effects on the epidemiology. As previously demonstrated, subsequent infections by two closely related viruses, such as those belonging to the Japanese encephalitis virus (JEV) serocomplex, can lead to partial or complete cross-immunity, altering the risk of infections or the outcome of disease. Two flaviviruses that may interact at population level are West Nile virus (WNV) and Usutu virus (USUV). These pathogens have antigenic cross-reactivity and affect human and animal populations throughout Europe. This systematic review investigates the overlap of WNV and USUV transmission cycles, not only geographically but also in terms of host and vector ranges. Co-circulation of WNV and USUV was reported in 10 countries and the viruses were found to infect 34 common bird species belonging to 11 orders. Moreover, four mosquito species are potential vectors for both viruses. Taken together, these data suggest that WNV and USUV transmission overlaps substantially in Europe and highlight the importance of further studies investigating the interactions between the two viruses within host and vector populations.

[Usutu virus: a novel flavivirus in Croatia]. / Usutski virus: novi flavivirus u hrvatskoj.

Usutu virus (USUV) belongs to the family Flaviviridae, genus Flavivirus, Japanese encephalitis serocomplex. The virus was discovered in 1959 in South Africa and has emerged since 1996 causing epizootics with high avian mortality in Europe. The importance of USUV in humans is not fully understood. However, several human clinical cases of USUV infection described so far indicate the role of this virus as an antropozoonotic agent. In Croatia, serologic evidence of USUV was first documented in 2011 in two horses from Zagreb and Sisak-Moslavina County. In 2012, USUV neutralizing antibodies were found in one human sample from a resident of a Vukovar-Srijem County. Human clinical cases of USUV infection were detected for the first time during the West Nile virus outbreak from July to September 2013. Three patients with USUV neuroinvasive disease were detected in the City of Zagreb and Zagreb County. Our results indicate USUV circulation in Croatia. Further human cases could be expected in the next transmission seasons.

An investigation on the diversity of mosquitocidal bacteria and its relationship with incidence of vector borne diseases.

Control of mosquitoes is the most important aspect of public health, as mosquitoes transmit many human diseases, including the fatal infection, Japanese encephalitis. This paper addresses the isolation of new mosquitocidal bacteria from soil samples in the Union Territory of Pondicherry, India, where, no clinical cases of vector borne infections have been reported. Bacterial isolates from soil samples were screened for potential mosquitocidal strains and bioassays against mosquito vectors (Culex quinquefasciatus, Anopheles stephensi and Aedes aegypti) were carried out. Genomic DNA of potential mosquitocidal isolates was amplified and species identification was carried out using BLASTn program (NCBI). Phylogenetic analysis of 16S rRNA sequences of mosquitocidal bacteria revealed seven potential isolates. SDS-PAGE results have shown that there was considerable difference in the protein profiles. Numerical analysis revealed 4 distinct groups at similarity level 25%. The relationship between VBDs and prevalence of soil mosquitocidal bacteria in the study sites has elicited considerable interest in the diversity of mosquitocidal bacteria and their application for mosquito borne diseases control.