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Topic:Mosquito-borne Diseases

Mosquito-borne diseases in horses encompass a range of illnesses transmitted by mosquito vectors, impacting equine health worldwide. These diseases include West Nile Virus (WNV), Eastern Equine Encephalitis (EEE), Western Equine Encephalitis (WEE), and Venezuelan Equine Encephalitis (VEE). Each disease is caused by distinct viral pathogens, leading to varying clinical presentations, such as fever, neurological symptoms, and in severe cases, death. Understanding the epidemiology, transmission dynamics, and pathogenesis of these diseases is essential for effective prevention and control strategies. This page compiles peer-reviewed research studies and scholarly articles that explore the transmission, clinical manifestations, and management of mosquito-borne diseases in horses.
No Evidence of Mosquito Involvement in the Transmission of Equine Hepacivirus (Flaviviridae) in an Epidemiological Survey of Austrian Horses.
Viruses    November 1, 2019   Volume 11, Issue 11 1014 doi: 10.3390/v11111014
Badenhorst M, de Heus P, Auer A, Rümenapf T, Tegtmeyer B, Kolodziejek J, Nowotny N, Steinmann E, Cavalleri JV.Prevalence studies have demonstrated a global distribution of equine hepacivirus (EqHV), a member of the family Flaviviridae. However, apart from a single case of vertical transmission, natural routes of EqHV transmission remain elusive. Many known flaviviruses are horizontally transmitted between hematophagous arthropods and vertebrate hosts. This study represents the first investigation of potential EqHV transmission by mosquitoes. More than 5000 mosquitoes were collected across Austria and analyzed for EqHV ribonucleic acid (RNA) by reverse transcription quantitative polymerase chain reacti...
West Nile fever in Europe in 2018: an emerging problem or just an anomaly?
The Veterinary record    September 29, 2019   Volume 185, Issue 12 365-368 doi: 10.1136/vr.l5748
Browne C, Medlock JM.Last summer saw an unusually high number of cases of West Nile fever in horses and people in south and south-east Europe, but it is too early to tell if this was a one-off increase or a sign of things to come. Here, , , and discuss the various West Nile fever surveillance and control mechanisms in place in the UK.
West Nile Virus: Veterinary Health and Vaccine Development.
Journal of medical entomology    September 25, 2019   Volume 56, Issue 6 1463-1466 doi: 10.1093/jme/tjz125
Bosco-Lauth AM, Bowen RA.West Nile virus (WNV) (Flaviviridae: Flavivirus) was discovered in Africa more than 80 yr ago and became recognized as an avian pathogen and a cause of neurologic disease in horses largely during periodic incursions into Europe. Introduction of WNV into North America stimulated great anxiety, particularly in the equine industry, but also for pet owners and livestock producers concerned about the effect of WNV on other domestic animals. Numerous subsequent studies of naturally occurring and experimentally induced disease greatly expanded our understanding of the host range and clinical conseque...
Equine viral encephalitis: prevalence, impact, and management strategies.
Veterinary medicine (Auckland, N.Z.)    August 7, 2019   Volume 10 99-110 doi: 10.2147/VMRR.S168227
Barba M, Fairbanks EL, Daly JM.Members of several different virus families cause equine viral encephalitis, the majority of which are arthropod-borne viruses (arboviruses) with zoonotic potential. The clinical signs caused are rarely pathognomonic; therefore, a clinical diagnosis is usually presumptive according to the geographical region. However, recent decades have seen expansion of the geographical range and emergence in new regions of numerous viral diseases. In this context, this review presents an overview of the prevalence and distribution of the main viral causes of equine encephalitis and discusses their impact an...
Can Attractive Sticky Traps Be Used to Protect Horses From the Bites of Stomoxys calcitrans (L.) (Diptera: Muscidae).
Journal of economic entomology    June 23, 2019   Volume 112, Issue 5 2469-2473 doi: 10.1093/jee/toz134
Tam TL, Hogsette J, TenBroeck S.The stable fly, Stomoxys calcitrans (L.), is a bloodsucking ectoparasite that causes irritation and distress to livestock, wildlife, and humans. Both sexes are vicious blood-feeders that feed on a variety of animals. Optically attractive sticky traps have been used to capture stable flies, and some companies claim that sticky traps can protect animals from the bites of stable flies. To further investigate the protective ability of sticky traps, Home and Garden Mosquito (HGM) traps were selected for evaluations at the University of Florida Horse Teaching Unit (HTU). Broodmares coated with fluor...
Emergence of Getah Virus Infection in Horse With Fever in China, 2018.
Frontiers in microbiology    June 20, 2019   Volume 10 1416 doi: 10.3389/fmicb.2019.01416
Lu G, Ou J, Ji J, Ren Z, Hu X, Wang C, Li S.Getah virus (GETV) is a mosquito-borne virus that was first determined in Malaysia in 1955, and can infect humans and multiple other mammals. GETV infection in horses has been reported in Japan and India, and causes great economic losses. In China, GETV has been identified in mosquitoes, pigs, foxes, and cattle with a wide geographical distribution, but has not been detected in horses. In August 2018, a sudden onset of fever was observed in racehorse in an equestrian training center in Guangdong Province in southern China. Blood samples were collected from the sick horse, and PCR/RT-PCR analys...
Associations between the presence of specific antibodies to the West Nile Virus infection and candidate genes in Romanian horses from the Danube delta.
Molecular biology reports    June 7, 2019   Volume 46, Issue 4 4453-4461 doi: 10.1007/s11033-019-04900-w
Stejskalova K, Janova E, Horecky C, Horecka E, Vaclavek P, Hubalek Z, Relling K, Cvanova M, D'Amico G, Mihalca AD, Modry D, Knoll A, Horin P.The West Nile virus (WNV) is a mosquito-borne flavivirus causing meningoencephalitis in humans and animals. Due to their particular susceptibility to WNV infection, horses serve as a sentinel species. In a population of Romanian semi-feral horses living in the Danube delta region, we have analyzed the distribution of candidate polymorphic genetic markers between anti WNV-IgG seropositive and seronegative horses. Thirty-six SNPs located in 28 immunity-related genes and 26 microsatellites located in the MHC and LY49 complex genomic regions were genotyped in 57 seropositive and 32 seronegative ho...
Culex torrentium: A Potent Vector for the Transmission of West Nile Virus in Central Europe.
Viruses    May 29, 2019   Volume 11, Issue 6 doi: 10.3390/v11060492
Jansen S, Heitmann A, Lühken R, Leggewie M, Helms M, Badusche M, Rossini G, Schmidt-Chanasit J, Tannich E.The continuous circulation of West Nile virus (WNV) in Central, South and East Europe and its recent detection in several dead birds and two horses in Germany highlights the need for information on WNV vector competence of mosquitoes from Central Europe. Therefore, three common Culex species (Culex pipiens biotype pipiens, Culex pipiens biotype molestus and Culex torrentium) from Germany were orally infected with WNV and kept at 18 °C, 21 °C, 24 °C or 27 °C for 14 or 21 days post infection (dpi). Thereafter viable WNV was present in the saliva in all tested taxa, but only at incubation tem...
Clinical and epidemiological features of West Nile virus equine encephalitis in New South Wales, Australia, 2011.
Australian veterinary journal    April 27, 2019   Volume 97, Issue 5 133-143 doi: 10.1111/avj.12810
Read AJ, Finlaison DS, Gu X, Hick PM, Moloney BJ, Wright T, Kirkland PD.Between February and June 2011, more than 300 horses with unexplained neurological disease were observed in New South Wales, Australia. A virulent strain of West Nile virus (WNV ), of Australian origin, was shown to be the cause of many of these cases. Methods: We reviewed the clinical descriptions provided by veterinary practitioners and the associated laboratory results. Although there was a range of clinical signs described, ataxia was the only sign that was consistently described in laboratory-confirmed cases. Results: WNV was detected in brain samples by real-time reverse transcription PC...
One Health approach for West Nile virus surveillance in the European Union: relevance of equine data for blood safety.
Euro surveillance : bulletin Europeen sur les maladies transmissibles = European communicable disease bulletin    April 25, 2019   Volume 24, Issue 16 1800349 doi: 10.2807/1560-7917.ES.2019.24.16.1800349
Young JJ, Coulombier D, Domanović D, Zeller H, Gossner CM.West Nile virus (WNV) infection is notifiable in humans and equids in the European Union (EU). An area where a human case is detected is considered affected until the end of the mosquito transmission season (week 48) and blood safety measures have to be implemented. We used human and equine case notifications between 2013 and 2017 to define the WNV distribution in the EU and to investigate the relevance of using equine cases as a complementary trigger for blood safety measures. Adding areas with equine cases to the definition of an affected area would have a major impact on blood safety measur...
Genetic susceptibility to West Nile virus infection in Camargue horses.
Research in veterinary science    April 10, 2019   Volume 124 284-292 doi: 10.1016/j.rvsc.2019.04.004
Stejskalova K, Cvanova M, Oppelt J, Janova E, Horecky C, Horecka E, Knoll A, Leblond A, Horin P.West Nile virus (WNV) is a mosquito-borne zoonotic neurotropic virus capable to cause lethal meningoencephalitis (WNE) in infected hosts such as birds, horses, and humans. Due to their sensitivity, horses serve as sentinel species in areas at risk. We studied a population of Camargue horses living in Southern France in two zones with endemic WNV circulation where WNV outbreaks were recorded in 2000 and 2003-4. Two sets of microsatellite markers located in MHC and Ly49 genomic regions were genotyped as well as multiple SNPs in ten immunity-related candidate gene regions. Associations between ge...
Serological evidence of infection with dengue and Zika viruses in horses on French Pacific Islands.
PLoS neglected tropical diseases    February 7, 2019   Volume 13, Issue 2 e0007162 doi: 10.1371/journal.pntd.0007162
Beck C, Leparc-Goffart I, Desoutter D, Debergé E, Bichet H, Lowenski S, Dumarest M, Gonzalez G, Migné C, Vanhomwegen J, Zientara S, Durand B....New Caledonia and French Polynesia are areas in which arboviruses circulate extensively. A large serological survey among horses from New Caledonia and French Polynesia was carried out to investigate the seroprevalence of flaviviruses in the horse population. Here, 293 equine sera samples were screened for flaviviruses using a competitive enzyme-linked immunosorbent assay (cELISA). The positive sera were then confirmed using a flavivirus-specific microsphere immunoassay (MIA) and seroneutralization tests. This serosurvey showed that 16.6% (27/163) and 30.8% (40/130) of horses were positive for...
Continued Evidence of Decline in the Enzootic Activity of Western Equine Encephalitis Virus in Colorado.
Journal of medical entomology    December 12, 2018   Volume 56, Issue 2 584-588 doi: 10.1093/jme/tjy214
Robb LL, Hartman DA, Rice L, deMaria J, Bergren NA, Borland EM, Kading RC.Western equine encephalitis (WEE) was once prevalent and routinely isolated from mosquitoes in Colorado; however, isolations of Western equine encephalitis virus (WEEV) have not been reported from mosquito pools since the early 1990s. The objective of the present study was to test pools of Culex tarsalis (Coquillett) mosquitoes sampled from Weld County, CO, in 2016 for evidence of WEEV infection. Over 7,000 mosquitoes were tested, but none were positive for WEEV RNA. These data indicate that WEEV either was not circulating enzootically in Northern Colorado, was very rare, and would require muc...
West Nile virus in horses – What do you need to know to diagnose the disease?
The Canadian veterinary journal = La revue veterinaire canadienne    December 5, 2018   Volume 59, Issue 10 1119-1120 
Paré J, Moore A.No abstract available
West Nile virus infection in horses in Saudi Arabia (in 2013-2015).
Zoonoses and public health    November 6, 2018   Volume 66, Issue 2 248-253 doi: 10.1111/zph.12532
Hemida MG, Perera RAPM, Chu DKW, Ko RLW, Alnaeem AA, Peiris M.West Nile virus (WNV) is an important emerging zoonotic arbovirus giving rise to clinical syndromes of varying severity in humans and horses. Culex mosquitoes are the main vector. Although WNV has been reported in many countries in the Middle East and Asia, little is known about its prevalence in equine populations in the Arabian Peninsula. We have carried out a serological study on 200 horses to assess WNV infection in the Eastern and Central regions of Saudi Arabia in 2013-2015. Sera were tested for the presence of WNV antibodies in parallel using a commercial enzyme-linked immunosorbent ass...
West Nile virus associated with equid encephalitis in Brazil, 2018.
Transboundary and emerging diseases    November 2, 2018   Volume 66, Issue 1 445-453 doi: 10.1111/tbed.13043
Silva ASG, Matos ACD, da Cunha MACR, Rehfeld IS, Galinari GCF, Marcelino SAC, Saraiva LHG, Martins NRDS, Maranhão RPA, Lobato ZIP, Pierezan F....Mosquito-borne arboviruses are a major public health concern worldwide and are responsible for emerging and re-emerging diseases. Taken together, the arboviruses have a strong impact on public health and are the most common causes of equine encephalitis. In-depth diagnostic investigation of equine viral encephalitis is of utmost importance for the epidemiological surveillance and control of this disease. Regarding neurological disorders in equids, in April-May 2018, at least 12 cases of equid mortality with acute neurological signs were reported in six farms from Espirito Santo state, Brazil. ...
Mammals repel mosquitoes with their tails.
The Journal of experimental biology    October 15, 2018   Volume 221, Issue Pt 20 jeb178905 doi: 10.1242/jeb.178905
Matherne ME, Cockerill K, Zhou Y, Bellamkonda M, Hu DL.The swinging of a mammal's tail has long been thought to deter biting insects, which, in cows, can drain up to 0.3 liters of blood per day. How effective is a mammal's tail at repelling insects? In this combined experimental and theoretical study, we filmed horses, zebras, elephants, giraffes and dogs swinging their tails. The tail swings at triple the frequency of a gravity-driven pendulum, and requires 27 times more power input. Tails can also be used like a whip to directly strike at insects. This whip-like effect requires substantial torques from the base of the tail on the order of 10-1...
In silico identification of immunodominant B-cell and T-cell epitopes of non-structural proteins of Usutu Virus.
Microbial pathogenesis    September 11, 2018   Volume 125 129-143 doi: 10.1016/j.micpath.2018.09.019
Satyam R, Janahi EM, Bhardwaj T, Somvanshi P, Haque S, Najm MZ.Usutu Virus (USUV; flavivirus) is a re-emerging pathogen invading the territories of European countries, Asia, and Africa. It is a mosquito-borne zoonotic virus with a bi-directional transmission route from animal to human and vice versa, and causes neurological disorders such as meningoencephalitis in bats, Homo sapiens, birds and horses. Due to limited availability of information about USUV and its deleterious effects on neural cells causing neurologic impairments, it becomes imperative to study this virus in detail to equip ourselves with a solution beforehand. The current study aims to ide...
Population genomics of Culiseta melanura, the principal vector of Eastern equine encephalitis virus in the United States.
PLoS neglected tropical diseases    August 17, 2018   Volume 12, Issue 8 e0006698 doi: 10.1371/journal.pntd.0006698
Soghigian J, Andreadis TG, Molaei G.Eastern Equine Encephalitis (EEE) (Togaviridae, Alphavirus) is a highly pathogenic mosquito-borne arbovirus that circulates in an enzootic cycle involving Culiseta melanura mosquitoes and wild Passeriformes birds in freshwater swamp habitats. Recently, the northeastern United States has experienced an intensification of virus activity with increased human involvement and northward expansion into new regions. In addition to its principal role in enzootic transmission of EEE virus among avian hosts, recent studies on the blood-feeding behavior of Cs. melanura throughout its geographic range sugg...
Seroprevalence and associated risk factors of mosquito-borne alphaviruses in horses in northern Queensland.
Australian veterinary journal    June 27, 2018   Volume 96, Issue 7 243-251 doi: 10.1111/avj.12711
Gummow B, Tan R, Joice RK, Burgess G, Picard J.To investigate the seroprevalence and associated risk factors of alphaviruses (Ross River virus (RRV), Barmah Forest virus (BFV) and Whataroa virus (WHAV)) in northern Queensland horses. Methods: A cross-sectional study of alphavirus antibodies in horses (n = 287) from 147 properties in northern Queensland from September 2013 to June 2014 was conducted. Owners of sampled horses were interviewed on potential risk factors. Data were analysed for associations using multivariable logistic regression. Results: Antibody titres for RRV were demonstrated in samples from 134 properties (91%; 95% co...
Large-Scale Complete-Genome Sequencing and Phylodynamic Analysis of Eastern Equine Encephalitis Virus Reveals Source-Sink Transmission Dynamics in the United States.
Journal of virology    May 29, 2018   Volume 92, Issue 12 e00074-18 doi: 10.1128/JVI.00074-18
Tan Y, Lam TT, Heberlein-Larson LA, Smole SC, Auguste AJ, Hennigan S, Halpin RA, Fedorova N, Puri V, Stockwell TB, Shilts MH, Andreadis T....Eastern equine encephalitis virus (EEEV) has a high case-fatality rate in horses and humans, and Florida has been hypothesized to be the source of EEEV epidemics for the northeastern United States. To test this hypothesis, we sequenced complete genomes of 433 EEEV strains collected within the United States from 1934 to 2014. Phylogenetic analysis suggested EEEV evolves relatively slowly and that transmission is enzootic in Florida, characterized by higher genetic diversity and long-term local persistence. In contrast, EEEV strains in New York and Massachusetts were characterized by lower genet...
Japanese encephalitis in Malaysia: An overview and timeline.
Acta tropica    May 29, 2018   Volume 185 219-229 doi: 10.1016/j.actatropica.2018.05.017
Kumar K, Arshad SS, Selvarajah GT, Abu J, Toung OP, Abba Y, Yasmin AR, Bande F, Sharma R, Ong BL.Japanese encephalitis (JE) is a vector-borne zoonotic disease caused by the Japanese encephalitis virus (JEV). It causes encephalitis in human and horses, and may lead to reproductive failure in sows. The first human encephalitis case in Malaya (now Malaysia) was reported during World War II in a British prison in 1942. Later, encephalitis was observed among race horses in Singapore. In 1951, the first JEV was isolated from the brain of an encephalitis patient. The true storyline of JE exposure among humans and animals has not been documented in Malaysia. In some places such as Sarawak, JEV ha...
Survey of UK horse owners’ knowledge of equine arboviruses and disease vectors.
The Veterinary record    May 15, 2018   Volume 183, Issue 5 159 doi: 10.1136/vr.104521
Chapman GE, Baylis M, Archer DC.Increased globalisation and climate change have led to concern about the increasing risk of arthropod-borne virus (arbovirus) outbreaks globally. An outbreak of equine arboviral disease in northern Europe could impact significantly on equine welfare, and result in economic losses. Early identification of arboviral disease by horse owners may help limit disease spread. In order to determine what horse owners understand about arboviral diseases of horses and their vectors, the authors undertook an open, cross-sectional online survey of UK horse owners. The questionnaire was distributed using soc...
Validation of a Risk Index Model for Predicting Eastern Equine Encephalitis Virus Transmission to Horses in Florida.
Journal of medical entomology    May 4, 2018   Volume 55, Issue 5 1143-1149 doi: 10.1093/jme/tjy067
Downs J, Vaziri M, Jenkins A, Unnasch T.Eastern Equine Encephalitis Virus (EEEV) is the most pathogenic arbovirus endemic to the United States. EEEV primarily infects birds but can be fatal to humans, horses, and some other mammals. Although EEEV transmission occurs in the Northeastern, Southeastern, and Midwestern United States, the largest number of horse and human cases have been reported in Florida, the only state where transmission occurs year round. Currently, a GIS-based risk index (RI) model is used to map EEE transmission risk to horses in Florida. This study validates that RI model using a 5-yr dataset of horse cases in Fl...
The challenges posed by equine arboviruses.
Equine veterinary journal    April 14, 2018   Volume 50, Issue 4 436-445 doi: 10.1111/evj.12829
Chapman GE, Baylis M, Archer D, Daly JM.Equine populations worldwide are at increasing risk of infection by viruses transmitted by biting arthropods, including mosquitoes, biting midges (Culicoides), sandflies and ticks. These include the flaviviruses (Japanese encephalitis, West Nile and Murray Valley encephalitis), alphaviruses (eastern, western and Venezuelan encephalitis) and the orbiviruses (African horse sickness and equine encephalosis). This review provides an overview of the challenges faced in the surveillance, prevention and control of the major equine arboviruses, particularly in the context of these viruses emerging in ...
West Nile virus outbreak in humans and epidemiological surveillance, west Andalusia, Spain, 2016.
Euro surveillance : bulletin Europeen sur les maladies transmissibles = European communicable disease bulletin    April 12, 2018   Volume 23, Issue 14 17-00261 doi: 10.2807/1560-7917.ES.2018.23.14.17-00261
López-Ruiz N, Montaño-Remacha MDC, Durán-Pla E, Pérez-Ruiz M, Navarro-Marí JM, Salamanca-Rivera C, Miranda B, Oyonarte-Gómez S....In Andalusia, Spain, West Nile virus (WNV) surveillance takes place from April to November, during the active vector period. Within this area seroconversion to this virus was evidenced in wild birds in 2004, affecting horses and two humans for the first time in 2010. Since 2010, the virus has been isolated every year in horses, and national and regional surveillance plans have been updated with the epidemiological changes found. WNV is spreading rapidly throughout southern Europe and has caused outbreaks in humans. Here we describe the second WNV outbreak in humans in Andalusia, with three con...
The Immune Responses of the Animal Hosts of West Nile Virus: A Comparison of Insects, Birds, and Mammals.
Frontiers in cellular and infection microbiology    April 3, 2018   Volume 8 96 doi: 10.3389/fcimb.2018.00096
Ahlers LRH, Goodman AG.Vector-borne diseases, including arboviruses, pose a serious threat to public health worldwide. Arboviruses of the flavivirus genus, such as Zika virus (ZIKV), dengue virus, yellow fever virus (YFV), and West Nile virus (WNV), are transmitted to humans from insect vectors and can cause serious disease. In 2017, over 2,000 reported cases of WNV virus infection occurred in the United States, with two-thirds of cases classified as neuroinvasive. WNV transmission cycles through two different animal populations: birds and mosquitoes. Mammals, particularly humans and horses, can become infected thro...
Alphaviruses: Serological Evidence of Human Infection in Paraguay (2012-2013).
Vector borne and zoonotic diseases (Larchmont, N.Y.)    March 13, 2018   Volume 18, Issue 5 266-272 doi: 10.1089/vbz.2017.2178
Alphaviruses can produce febrile illness and encephalitis in dead-end hosts such as horses and humans. Within this genus, the Venezuelan Equine Encephalitis virus (VEEV) complex includes pathogenic epizootic subtypes and enzootic subtypes that are not pathogenic in horses (except subtype IE, Mexican strains), although they can cause febrile symptoms in humans. The Rio Negro virus (RNV-VEEV subtype VI) circulates in Argentina, where it was associated with undifferentiated febrile illness. Mayaro (MAYV) and Una (UNAV) viruses belong to a different group, the Semliki Forest virus complex, with co...
A Survey on West Nile and Usutu Viruses in Horses and Birds in Poland.
Viruses    February 17, 2018   Volume 10, Issue 2 87 doi: 10.3390/v10020087
Bażanów B, Jansen van Vuren P, Szymański P, Stygar D, Frącka A, Twardoń J, Kozdrowski R, Pawęska JT.West Nile virus (WNV) and Usutu virus (USUV) are members of the family Flaviviridae which, natural life cycles involve mosquito-bird-mosquito transmission. Both represent emerging viruses in Europe with potential to cause neuroinvasive disease in humans. This study investigates the seroprevalence of serum neutralizing antibodies to WNV and to USUV in birds and in horses in Poland. Antibodies against WNV and USUV were detected in 5 (35.7%) and in 1 (7.14%) of 14 birds and in 62 (15.08%) and in 115 (27.98%) of 411 horses, respectively. Twenty-one WNV serologically positive horses (33.87%) and 67...
A Serosurvey of Flavivirus Infection in Horses and Birds in Slovakia.
Vector borne and zoonotic diseases (Larchmont, N.Y.)    February 13, 2018   Volume 18, Issue 4 206-213 doi: 10.1089/vbz.2017.2216
Csank T, Drzewnioková P, Korytár Ľ, Major P, Gyuranecz M, Pistl J, Bakonyi T.In central Europe, at least three flaviviruses circulate among vectors and vertebrate hosts. West Nile virus (WNV) and Usutu virus (USUV) are mosquito-borne viruses maintained in the nature by enzootic cycle between mosquitoes and birds. Tick-borne encephalitis virus (TBEV) is a flavivirus causing annual human cases in Slovakia. The aim of this study is the prevalence assessment of flavivirus infections in horses (n = 145) and birds (n = 109) by enzyme-linked immunosorbent assay (ELISA) and confirmation by neutralization test (VNT). WNV antibodies have been detected in 11.7% of tested ...
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