First Detection of West Nile Virus Lineage 2 in <i>Culex pipiens</i> Vectors in Croatia

The West Nile virus (WNV) has recently become more widespread, posing a threat to both human and animal health. In Western Europe, most outbreaks have been caused by WNV lineage 1, while in Eastern Europe, WNV lineage 2 has led to human and bird mortality. The ability to appropriately manage this th...

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Main Authors: Goran Vignjević, Nataša Bušić, Nataša Turić, Zsaklin Varga, Brigitta Zana, Ágota Ábrahám, Kornélia Kurucz, Ivana Vrućina, Enrih Merdić
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Language:English
Published: MDPI AG 2024-12-01
Series:Pathogens
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Online Access:https://www.mdpi.com/2076-0817/13/12/1131
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author Goran Vignjević
Nataša Bušić
Nataša Turić
Zsaklin Varga
Brigitta Zana
Ágota Ábrahám
Kornélia Kurucz
Ivana Vrućina
Enrih Merdić
author_facet Goran Vignjević
Nataša Bušić
Nataša Turić
Zsaklin Varga
Brigitta Zana
Ágota Ábrahám
Kornélia Kurucz
Ivana Vrućina
Enrih Merdić
author_sort Goran Vignjević
collection DOAJ
description The West Nile virus (WNV) has recently become more widespread, posing a threat to both human and animal health. In Western Europe, most outbreaks have been caused by WNV lineage 1, while in Eastern Europe, WNV lineage 2 has led to human and bird mortality. The ability to appropriately manage this threat is dependent on integrated surveillance and early detection. This study aimed to quantify the prevalence of WNV infection in mosquitoes and to identify the circulating viral lineage in eastern Croatia. Mosquito traps were set up in rural and urban areas during the 2021–2023 seasons, and the collected specimens were identified morphologically. Mosquito species <i>Culex pipiens</i> and <i>Aedes albopictus</i> were tested for Flaviviruses using conventional PCR in a heminested system. The positive samples were then subjected to a specific real-time PCR designed to detect WNV. A total of 385 mosquito pools were tested, and positive pools were found in samples from Osijek-Baranja and Vukovar-Srijem, both of which contained <i>Cx. pipiens</i> mosquitoes. Sequencing of amplicons revealed WNV lineage 2 partial NS5 gene sequences. Phylogenetic analysis suggests the Hungarian origin of strain, which complements birds’ migratory routes. These findings indicate the first detection of WNV in mosquitoes in Croatia. This suggests that human cases in this region are likely due to infections with lineage 2 transmitted by local <i>Culex</i> mosquitoes.
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spelling doaj-art-ba04e7ab014f45eb9b06a145efdba7252025-08-20T02:56:59ZengMDPI AGPathogens2076-08172024-12-011312113110.3390/pathogens13121131First Detection of West Nile Virus Lineage 2 in <i>Culex pipiens</i> Vectors in CroatiaGoran Vignjević0Nataša Bušić1Nataša Turić2Zsaklin Varga3Brigitta Zana4Ágota Ábrahám5Kornélia Kurucz6Ivana Vrućina7Enrih Merdić8Department of Biology, University Josip Juraj Strossmayer of Osijek, 31000 Osijek, CroatiaDepartment of Biology, University Josip Juraj Strossmayer of Osijek, 31000 Osijek, CroatiaDepartment of Biology, University Josip Juraj Strossmayer of Osijek, 31000 Osijek, CroatiaNational Laboratory of Virology, Szentagothai Research Centre, University of Pécs, 7600 Pécs, HungaryNational Laboratory of Virology, Szentagothai Research Centre, University of Pécs, 7600 Pécs, HungaryNational Laboratory of Virology, Szentagothai Research Centre, University of Pécs, 7600 Pécs, HungaryNational Laboratory of Virology, Szentagothai Research Centre, University of Pécs, 7600 Pécs, HungaryDepartment of Biology, University Josip Juraj Strossmayer of Osijek, 31000 Osijek, CroatiaDepartment of Biology, University Josip Juraj Strossmayer of Osijek, 31000 Osijek, CroatiaThe West Nile virus (WNV) has recently become more widespread, posing a threat to both human and animal health. In Western Europe, most outbreaks have been caused by WNV lineage 1, while in Eastern Europe, WNV lineage 2 has led to human and bird mortality. The ability to appropriately manage this threat is dependent on integrated surveillance and early detection. This study aimed to quantify the prevalence of WNV infection in mosquitoes and to identify the circulating viral lineage in eastern Croatia. Mosquito traps were set up in rural and urban areas during the 2021–2023 seasons, and the collected specimens were identified morphologically. Mosquito species <i>Culex pipiens</i> and <i>Aedes albopictus</i> were tested for Flaviviruses using conventional PCR in a heminested system. The positive samples were then subjected to a specific real-time PCR designed to detect WNV. A total of 385 mosquito pools were tested, and positive pools were found in samples from Osijek-Baranja and Vukovar-Srijem, both of which contained <i>Cx. pipiens</i> mosquitoes. Sequencing of amplicons revealed WNV lineage 2 partial NS5 gene sequences. Phylogenetic analysis suggests the Hungarian origin of strain, which complements birds’ migratory routes. These findings indicate the first detection of WNV in mosquitoes in Croatia. This suggests that human cases in this region are likely due to infections with lineage 2 transmitted by local <i>Culex</i> mosquitoes.https://www.mdpi.com/2076-0817/13/12/1131VBDtiger mosquito<i>Aedes albopictus</i>common house mosquito<i>Culex pipiens</i>arbovirus
spellingShingle Goran Vignjević
Nataša Bušić
Nataša Turić
Zsaklin Varga
Brigitta Zana
Ágota Ábrahám
Kornélia Kurucz
Ivana Vrućina
Enrih Merdić
First Detection of West Nile Virus Lineage 2 in <i>Culex pipiens</i> Vectors in Croatia
Pathogens
VBD
tiger mosquito
<i>Aedes albopictus</i>
common house mosquito
<i>Culex pipiens</i>
arbovirus
title First Detection of West Nile Virus Lineage 2 in <i>Culex pipiens</i> Vectors in Croatia
title_full First Detection of West Nile Virus Lineage 2 in <i>Culex pipiens</i> Vectors in Croatia
title_fullStr First Detection of West Nile Virus Lineage 2 in <i>Culex pipiens</i> Vectors in Croatia
title_full_unstemmed First Detection of West Nile Virus Lineage 2 in <i>Culex pipiens</i> Vectors in Croatia
title_short First Detection of West Nile Virus Lineage 2 in <i>Culex pipiens</i> Vectors in Croatia
title_sort first detection of west nile virus lineage 2 in i culex pipiens i vectors in croatia
topic VBD
tiger mosquito
<i>Aedes albopictus</i>
common house mosquito
<i>Culex pipiens</i>
arbovirus
url https://www.mdpi.com/2076-0817/13/12/1131
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