BACKGROUNDSerological evidence of West Nile virus (WNV) infection has been reported in different regions of Brazil from equine and human hosts but the virus had never been isolated in the country.OBJECTIVES We sought to identify the viral etiology of equine encephalitis in Espírito Santo state.METHODS We performed viral culture in C6/36 cells, molecular detection of WNV genome, histopathology and immunohistochemistry from horse cerebral tissue. We also carried out sequencing, phylogenetic analysis and molecular clock.FINDINGS Histopathologic analysis from horse cerebral tissue showed injury related to encephalitis and WNV infection was confirmed by immunohistochemistry. The virus was detected by reverse transcription quantitative polymerase chain reaction (RT-qPCR) from brain tissue and subsequently isolated in C6/36 cells. WNV full-length genome was sequenced showing the isolated strain belongs to lineage 1a. The molecular clock indicated that Brazilian WNV strain share the same common ancestor that were circulating in US during 2002-2005.MAIN CONCLUSIONS Here we report the first isolation of WNV in Brazil from a horse with neurologic disease, which was clustered into lineage 1a with others US WNV strains isolated in beginning of 2000’s decade.

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Many human arboviruses are also pathogenic for horses, and some of these have emerged recently. A descriptive cross-sectional observational study was conducted to assess the prevalence of West Nile virus (WNV) and other arboviruses among 77 horses on the rural properties of the Espirito Santo state, Brazil. Serum samples were screened for arbovirus-reactive antibodies using the hemagglutination inhibition technique and subsequently a plaque reduction neutralization test for the confirmation of exposure from sera was used to detect heterotypic immune reactions. Overall, the total antibodies against at least one arbovirus of Alphavirus, Flavivirus, and Orthobunyavirus genera were detected in 39 (50.6%) animals. The antibodies to Phlebovirus were not detected in any sample. When the 24 WNV hemagglutination inhibition (HI)-positive samples were tested by the plaque-reduction neutralization test 90%, 9 (32.1%) were positive for WNV antibodies and 14 (50%) for Saint Louis encephalitis virus. Our findings indicate that the region provides ideal conditions for the emergence of arboviruses, reinforcing the need for further surveillance of mosquito-transmitted diseases in domestic animals.