{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,12]],"date-time":"2026-06-12T20:23:10Z","timestamp":1781295790221,"version":"3.54.1"},"reference-count":40,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2023,1,13]],"date-time":"2023-01-13T00:00:00Z","timestamp":1673568000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"European Union\u2019s Horizon 2020 research and innovation program VEO","award":["874735"],"award-info":[{"award-number":["874735"]}]},{"name":"European Union\u2019s Horizon 2020 research and innovation program VEO","award":["\u03a42\u0395\u0394\u039a-02070"],"award-info":[{"award-number":["\u03a42\u0395\u0394\u039a-02070"]}]},{"name":"Operational Program for Competitiveness, Entrepreneurship, and Innovation project EMPROS","award":["874735"],"award-info":[{"award-number":["874735"]}]},{"name":"Operational Program for Competitiveness, Entrepreneurship, and Innovation project EMPROS","award":["\u03a42\u0395\u0394\u039a-02070"],"award-info":[{"award-number":["\u03a42\u0395\u0394\u039a-02070"]}]},{"name":"National Public Health Organization in Greece","award":["874735"],"award-info":[{"award-number":["874735"]}]},{"name":"National Public Health Organization in Greece","award":["\u03a42\u0395\u0394\u039a-02070"],"award-info":[{"award-number":["\u03a42\u0395\u0394\u039a-02070"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>In 2022, Greece was the second most seriously affected European country in terms of the West Nile virus (WNV), after Italy. Specifically, Central Macedonia was the region with the most reported human cases (81.5%). In the present study, 30,816 female Culex pipiens sensu lato mosquitoes were collected from May to September 2022 in the seven regional units of Central Macedonia; they were then grouped into 690 pools and tested for WNV, while next-generation sequencing was applied to the samples, which showed a cycle threshold of Ct &lt; 30 in a real-time RT-PCR test. WNV was detected in 5.9% of pools, with significant differences in the detection rate among regional units and months. It is of interest that in the Thessaloniki regional unit, where most of the human cases were observed, the virus circulation started earlier, peaked earlier, and lasted longer than in the other regional units. All sequences clustered into the Central European subclade of WNV lineage 2, and the virus strain differed from the initial Greek strain of 2010 by 0.52% and 0.27% at the nucleotide and amino acid levels, respectively. Signature substitutions were present, such as S73P and T157A in the prM and E structural proteins, respectively. The screening of mosquitoes provides useful information for virus circulation in a region with a potential for early warning, while the availability of whole-genome sequences is essential for further studies, including virus evolution.<\/jats:p>","DOI":"10.3390\/v15010224","type":"journal-article","created":{"date-parts":[[2023,1,13]],"date-time":"2023-01-13T04:02:04Z","timestamp":1673582524000},"page":"224","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":16,"title":["West Nile Virus in Culex Mosquitoes in Central Macedonia, Greece, 2022"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5200-8045","authenticated-orcid":false,"given":"Katerina","family":"Tsioka","sequence":"first","affiliation":[{"name":"Laboratory of Microbiology, National Reference Centre for Arboviruses, School of Medicine, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5236-9537","authenticated-orcid":false,"given":"Sandra","family":"Gewehr","sequence":"additional","affiliation":[{"name":"Ecodevelopment S.A., 57010 Thessaloniki, Greece"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Styliani","family":"Pappa","sequence":"additional","affiliation":[{"name":"Laboratory of Microbiology, National Reference Centre for Arboviruses, School of Medicine, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stella","family":"Kalaitzopoulou","sequence":"additional","affiliation":[{"name":"Ecodevelopment S.A., 57010 Thessaloniki, Greece"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Konstantina","family":"Stoikou","sequence":"additional","affiliation":[{"name":"Laboratory of Microbiology, National Reference Centre for Arboviruses, School of Medicine, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Spiros","family":"Mourelatos","sequence":"additional","affiliation":[{"name":"Ecodevelopment S.A., 57010 Thessaloniki, Greece"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6643-3322","authenticated-orcid":false,"given":"Anna","family":"Papa","sequence":"additional","affiliation":[{"name":"Laboratory of Microbiology, National Reference Centre for Arboviruses, School of Medicine, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,1,13]]},"reference":[{"key":"ref_1","first-page":"471","article-title":"A neurotropic virus isolated from the blood of a native of Uganda","volume":"20","author":"Smithburn","year":"1970","journal-title":"Am. J. Trop. Med. Hyg."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"811","DOI":"10.3390\/v3060811","article-title":"West Nile virus: Immunity and pathogenesis","volume":"3","author":"Lim","year":"2011","journal-title":"Viruses"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"571","DOI":"10.3109\/23744235.2016.1164890","article-title":"Epidemiological and clinical aspects on West Nile virus, a globally emerging pathogen","volume":"48","author":"David","year":"2016","journal-title":"Infect. Dis."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1016\/j.cll.2009.10.006","article-title":"West Nile virus","volume":"30","author":"Rossi","year":"2010","journal-title":"Clin. Lab. Med."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"171","DOI":"10.1016\/S1474-4422(07)70030-3","article-title":"West Nile virus","volume":"6","author":"Kramer","year":"2007","journal-title":"Lancet Neurol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"175","DOI":"10.1128\/microbiolspec.EI10-0021-2016","article-title":"West Nile Virus Infection","volume":"4","author":"Sejvar","year":"2016","journal-title":"Microbiol. Spectr."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Fall, G., Di Paola, N., Faye, M., Dia, M., Freire, C.C.M., Loucoubar, C., Zanotto, P.M.A., Faye, O., and Sall, A.A. (2017). Biological and phylogenetic characteristics of West African lineages of West Nile virus. PLoS Negl. Trop. Dis., 11.","DOI":"10.1371\/journal.pntd.0006078"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Casimiro-Soriguer, C.S., Perez-Florido, J., Fernandez-Rueda, J.L., Pedrosa-Corral, I., Guillot-Sulay, V., Lorusso, N., Martinez-Gonzalez, L.J., Navarro-Mari, J.M., Dopazo, J., and Sanbonmatsu-Gamez, S. (2021). Phylogenetic Analysis of the 2020 West Nile Virus (WNV) Outbreak in Andalusia (Spain). Viruses, 13.","DOI":"10.20944\/preprints202102.0604.v1"},{"key":"ref_9","first-page":"239","article-title":"[Epidemiology of the West Nile virus: Study of a focus in Camargue. IV. Meningo-encephalomyelitis of the horse]","volume":"118","author":"Joubert","year":"1970","journal-title":"Ann. Inst. Pasteur."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"618","DOI":"10.3201\/eid1204.051379","article-title":"Lineage 1 and 2 strains of encephalitic West Nile virus, central Europe","volume":"12","author":"Bakonyi","year":"2006","journal-title":"Emerg. Infect. Dis."},{"key":"ref_11","first-page":"29","article-title":"[Genotyping of West Nile fever virus strains circulating in southern Russia as an epidemiological investigation method: Principles and results]","volume":"2","author":"Platonov","year":"2011","journal-title":"Zh. Mikrobiol. Epidemiol. Immunobiol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"19644","DOI":"10.2807\/ese.15.34.19644-en","article-title":"Ongoing outbreak of West Nile virus infections in humans in Greece, July-August 2010","volume":"15","author":"Papa","year":"2010","journal-title":"Eurosurveillance"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1868","DOI":"10.3201\/eid1710.110525","article-title":"Outbreak of West Nile virus infection in Greece, 2010","volume":"17","author":"Danis","year":"2011","journal-title":"Emerg. Infect. Dis."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"920","DOI":"10.3201\/eid1705.101759","article-title":"Genetic characterization of West Nile virus lineage 2, Greece, 2010","volume":"17","author":"Papa","year":"2011","journal-title":"Emerg. Infect. Dis."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1176","DOI":"10.1111\/j.1469-0691.2010.03438.x","article-title":"Detection of West Nile virus lineage 2 in mosquitoes during a human outbreak in Greece","volume":"17","author":"Papa","year":"2011","journal-title":"Clin. Microbiol. Infect."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1900543","DOI":"10.2807\/1560-7917.ES.2020.25.32.1900543","article-title":"West Nile virus in humans, Greece, 2018: The largest seasonal number of cases, 9 years after its emergence in the country","volume":"25","author":"Pervanidou","year":"2020","journal-title":"Eurosurveillance"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"555","DOI":"10.1093\/trstmh\/tru100","article-title":"Detection of West Nile virus and insect-specific flavivirus RNA in Culex mosquitoes, central Macedonia, Greece","volume":"108","author":"Papa","year":"2014","journal-title":"Trans. R. Soc. Trop. Med. Hyg."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"19862","DOI":"10.2807\/ese.16.18.19862-en","article-title":"Molecular detection and phylogenetic analysis of West Nile virus lineage 2 in sedentary wild birds (Eurasian magpie), Greece, 2010","volume":"16","author":"Valiakos","year":"2011","journal-title":"Eurosurveillance"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"6534","DOI":"10.3390\/ijerph10126534","article-title":"West Nile virus state of the art report of MALWEST Project","volume":"10","author":"Marka","year":"2013","journal-title":"Int. J. Environ. Res. Public Health"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"e219","DOI":"10.1111\/tbed.12334","article-title":"Emergence of Equine West Nile Encephalitis in Central Macedonia, Greece, 2010","volume":"63","author":"Bouzalas","year":"2016","journal-title":"Transbound. Emerg. Dis."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Sofia, M., Giannakopoulos, A., Giantsis, I.A., Touloudi, A., Birtsas, P., Papageorgiou, K., Athanasakopoulou, Z., Chatzopoulos, D.C., Vrioni, G., and Galamatis, D. (2022). West Nile Virus Occurrence and Ecological Niche Modeling in Wild Bird Species and Mosquito Vectors: An Active Surveillance Program in the Peloponnese Region of Greece. Microorganisms, 10.","DOI":"10.3390\/microorganisms10071328"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"2323","DOI":"10.3201\/eid2512.181225","article-title":"Phylogenetic Analysis of Bird-Virulent West Nile Virus Strain, Greece","volume":"25","author":"Valiakos","year":"2019","journal-title":"Emerg. Infect. Dis."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"105278","DOI":"10.1016\/j.actatropica.2019.105278","article-title":"Detection of flaviviruses and alphaviruses in mosquitoes in Central Macedonia, Greece, 2018","volume":"202","author":"Papa","year":"2020","journal-title":"Acta Trop."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"106391","DOI":"10.1016\/j.actatropica.2022.106391","article-title":"Detection and molecular characterization of West Nile virus in Culex pipiens mosquitoes in Central Macedonia, Greece, 2019\u20132021","volume":"230","author":"Tsioka","year":"2022","journal-title":"Acta Trop."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"344","DOI":"10.1111\/zph.12152","article-title":"Surveillance and Early Warning of West Nile Virus Lineage 2 Using Backyard Chickens and Correlation to Human Neuroinvasive Cases","volume":"62","author":"Chaintoutis","year":"2015","journal-title":"Zoonoses Public Health"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1016\/j.cimid.2014.01.004","article-title":"Evaluation of a West Nile virus surveillance and early warning system in Greece, based on domestic pigeons","volume":"37","author":"Chaintoutis","year":"2014","journal-title":"Comp. Immunol. Microbiol. Infect. Dis."},{"key":"ref_27","unstructured":"European Centre for Disease Prevention and Control (2022, December 11). Weekly Updates: 2022 West Nile Virus Transmission Season. Available online: https:\/\/www.ecdc.europa.eu\/en\/west-nile-fever\/surveillance-and-disease-data\/disease-data-ecdc."},{"key":"ref_28","first-page":"247","article-title":"Keys for the identification of the mosquitoes of Greece","volume":"13","author":"Darsie","year":"1997","journal-title":"J. Am. Mosq. Control Assoc."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Becker, N., Petric, D., Zgomba, M., Boase, C., Madon, M., Dahl, C., and Kaiser, A. (2010). Mosquitoes and Their Control, Springer.","DOI":"10.1007\/978-3-540-92874-4"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"101774","DOI":"10.1016\/j.mcp.2021.101774","article-title":"PCR-based next-generation West Nile virus sequencing protocols","volume":"60","author":"Pappa","year":"2021","journal-title":"Mol. Cell. Probes"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"3022","DOI":"10.1093\/molbev\/msab120","article-title":"MEGA11: Molecular Evolutionary Genetics Analysis Version 11","volume":"38","author":"Tamura","year":"2021","journal-title":"Mol. Biol. Evol."},{"key":"ref_32","unstructured":"Biggerstaff, B. (2022). PooledInfRate, Version 4.0: An Excel\u00a9 Add-In to Compute Infection Rates from Pooled Data. Centers for Disease Control and Prevention."},{"key":"ref_33","unstructured":"National Public Health Organization (2022). Weekly Epidemiological Report for West Nile Virus disease, Greece, 06.12.2022."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1551","DOI":"10.1007\/s00436-013-3302-x","article-title":"West Nile virus in mosquitoes in Greece","volume":"112","author":"Papa","year":"2013","journal-title":"Parasitol. Res."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"60","DOI":"10.1089\/vbz.2018.2422","article-title":"Entomological Data and Detection of West Nile Virus in Mosquitoes in Greece (2014-2016), Before Disease Re-Emergence in 2017","volume":"20","author":"Patsoula","year":"2020","journal-title":"Vector Borne Zoonotic Dis."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Paz, S., Malkinson, D., Green, M.S., Tsioni, G., Papa, A., Danis, K., Sirbu, A., Ceianu, C., Katalin, K., and Ferenczi, E. (2013). Permissive summer temperatures of the 2010 European West Nile fever upsurge. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0056398"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Chauhan, B., Jabran, K., and Mahajan, G. (2017). Rice production in Europe. Rice Production Worldwide, Springer.","DOI":"10.1007\/978-3-319-47516-5"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"351","DOI":"10.1016\/j.jcv.2013.02.020","article-title":"West Nile virus infections in humans--focus on Greece","volume":"58","author":"Papa","year":"2013","journal-title":"J. Clin. Virol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"106010","DOI":"10.1016\/j.actatropica.2021.106010","article-title":"West Nile fever upsurge in a Greek regional unit, 2020","volume":"221","author":"Papa","year":"2021","journal-title":"Acta Trop."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1673","DOI":"10.1007\/s00705-019-04243-8","article-title":"Emergence of West Nile virus lineage 2 belonging to the Eastern European subclade, Greece","volume":"164","author":"Papa","year":"2019","journal-title":"Arch. Virol."}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/15\/1\/224\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T18:05:09Z","timestamp":1760119509000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/15\/1\/224"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,1,13]]},"references-count":40,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2023,1]]}},"alternative-id":["v15010224"],"URL":"https:\/\/doi.org\/10.3390\/v15010224","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,1,13]]}}}