355b0012e90a911e3dc1983505597fa9 pmc_11209238.pdf fe3c73e0480e16ec09c9be8635be301aedf6c9ca pmc_11209238.pdf 77bf3151faef772bb9b29543d07463e60c45df255cd4120dcd70e1e3a1fb1768 pmc_11209238.pdf Title: West Nile Virus Seroprevalence in Wild Birds and Equines in Madrid Province, Spain Subject: West Nile virus (WNV) is a re-emerging flavivirus, primarily circulating among avian hosts and mosquito vectors, causing periodic outbreaks in humans and horses, often leading to neuroinvasive disease and mortality. Spain has reported several outbreaks, most notably in 2020 with seventy-seven human cases and eight fatalities. WNV has been serologically detected in horses in the Community of Madrid, but to our knowledge, it has never been reported from wild birds in this region. To estimate the seroprevalence of WNV in wild birds and horses in the Community of Madrid, 159 wild birds at a wildlife rescue center and 25 privately owned equines were sampled. Serum from thirteen birds (8.2%) and one equine (4.0%) tested positive with a WNV competitive enzyme-linked immunosorbent assay (cELISA) designed for WNV antibody detection but sensitive to cross-reacting antibodies to other flaviviruses. Virus-neutralization test (VNT) confirmed WNV antibodies in four bird samples (2.5%), and antibodies to undetermined flavivirus in four additional samples. One equine sample (4.0%) tested positive for WNV by VNT, although this horse previously resided in a WN-endemic area. ELISA-positive birds included both migratory and resident species, juveniles and adults. Two seropositive juvenile birds suggest local flavivirus transmission within the Community of Madrid, while WNV seropositive adult birds may have been infected outside Madrid. The potential circulation of flaviviruses, including WNV, in birds in the Madrid Community raises concerns, although further surveillance of mosquitoes, wild birds, and horses in Madrid is necessary to establish the extent of transmission and the principal species involved. Keywords: flavivirus; West Nile virus; birds; equines; seroprevalence; central Spain Author: Richard A. J. Williams, Hillary A. Criollo Valencia, Irene López Márquez, Fernando González González, Francisco Llorente, Miguel Ángel Jiménez-Clavero, Núria Busquets, Marta Mateo Barrientos, Gustavo Ortiz-Díez and Tania Ayllón Santiago Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25 CreationDate: Fri Jun 7 07:58:33 2024 CEST ModDate: Fri Jun 7 08:07:20 2024 CEST Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 14 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 5394249 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- WIRPUU+VnURWPalladioL-Bold Type 1 Custom yes yes yes 10 0 YCRIOM+URWPalladioL-Bold Type 1 Custom yes yes yes 16 0 UMXSRO+URWPalladioL-Roma Type 1 Custom yes yes yes 22 0 IGPNRD+URWPalladioL-Ital Type 1 Custom yes yes yes 27 0 SAVOOC+VnURWPalladioL Type 1 Custom yes yes yes 32 0 CTCXDQ+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 48 0 XYPWHM+CMSY10 Type 1 Builtin yes yes yes 68 0 OJNCOJ+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 83 0 OJNCOL+PalatinoLinotype TrueType WinAnsi yes yes no 86 0 OJNCON+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 89 0 ICBUDW+URWPalladioL-BoldItal Type 1 Custom yes yes yes 99 0 OJNCOJ+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 114 0 OJNCOL+PalatinoLinotype TrueType WinAnsi yes yes no 117 0 OJNCON+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 120 0 CRUZTT+VnURWPalladioL-Italic Type 1 Custom yes yes yes 144 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2025-02-23 02:08:22 CET RepresentationInformation: pmc_11209238.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2025-02-22 02:06:45 CET Size: 5394249 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 379 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Materials and Methods Destination: section.2 Children: Item: Title: Samples Destination: subsection.2.1 Children: Item: Title: Birds Destination: subsubsection.2.1.1 Item: Title: Equines Destination: subsubsection.2.1.2 Item: Title: Serological Testing Destination: subsection.2.2 Item: Title: Statistical Analysis Destination: subsection.2.3 Item: Title: Results Destination: section.3 Children: Item: Title: Birds Destination: subsection.3.1 Item: Title: Equids Destination: subsection.3.2 Item: Title: Discussion Destination: section.4 Item: Title: References Destination: appendix.A. Info: Title: West Nile Virus Seroprevalence in Wild Birds and Equines in Madrid Province, Spain Author: Richard A. J. Williams, Hillary A. Criollo Valencia, Irene López Márquez, Fernando González González, Francisco Llorente, Miguel Ángel Jiménez-Clavero, Núria Busquets, Marta Mateo Barrientos, Gustavo Ortiz-Díez and Tania Ayllón Santiago Subject: West Nile virus (WNV) is a re-emerging flavivirus, primarily circulating among avian hosts and mosquito vectors, causing periodic outbreaks in humans and horses, often leading to neuroinvasive disease and mortality. Spain has reported several outbreaks, most notably in 2020 with seventy-seven human cases and eight fatalities. WNV has been serologically detected in horses in the Community of Madrid, but to our knowledge, it has never been reported from wild birds in this region. To estimate the seroprevalence of WNV in wild birds and horses in the Community of Madrid, 159 wild birds at a wildlife rescue center and 25 privately owned equines were sampled. Serum from thirteen birds (8.2%) and one equine (4.0%) tested positive with a WNV competitive enzyme-linked immunosorbent assay (cELISA) designed for WNV antibody detection but sensitive to cross-reacting antibodies to other flaviviruses. Virus-neutralization test (VNT) confirmed WNV antibodies in four bird samples (2.5%), and antibodies to undetermined flavivirus in four additional samples. One equine sample (4.0%) tested positive for WNV by VNT, although this horse previously resided in a WN-endemic area. ELISA-positive birds included both migratory and resident species, juveniles and adults. Two seropositive juvenile birds suggest local flavivirus transmission within the Community of Madrid, while WNV seropositive adult birds may have been infected outside Madrid. The potential circulation of flaviviruses, including WNV, in birds in the Madrid Community raises concerns, although further surveillance of mosquitoes, wild birds, and horses in Madrid is necessary to establish the extent of transmission and the principal species involved. 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