FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / Transbound Emerg Dis / First Detection of West Nile Virus (WNV) Lineage 2 in Mosqui...
Transboundary and emerging diseases2025; 2025; 3208806; doi: 10.1155/tbed/3208806

First Detection of West Nile Virus (WNV) Lineage 2 in Mosquitoes in the Republic of Kosovo.

Abstract: West Nile virus (WNV, family Flaviviridae) is the most geographically widespread arbovirus affecting humans. It circulates between wild birds and mosquitoes, while humans and horses are dead-end hosts. In recent years, several outbreaks have been reported from European countries, including the Balkan Peninsula. In the Republic of Kosovo, a southern Balkan country, data on WNV are scarce, and neither mosquito monitoring nor WNV surveillance is established. To address this gap, we aimed to assess a first monitoring approach that should set the basis and support future large-scale activities in the country. Mosquito sampling was performed from May to September 2022 in a peri-urban area in the western part of the capital city Prishtina, Republic of Kosovo. Collected mosquitoes were pooled, homogenized, and total nucleic acid was extracted. A WNV-DENV-ZIKV-specific multiplex RT-qPCR was applied, and WNV-positive samples were confirmed by RT-PCR and whole-genome sequencing. Of 44 screened pools, one pool molecularly identified as Culex pipiens f. pipiens was positive for WNV RNA. Subsequent sequencing revealed WNV lineage 2, and phylogenetic analysis included our sample in a monophyletic clade consisting mostly of sequences from southeastern Europe. This finding represents the first detection of WNV in mosquitoes in Kosovo, and provides crucial baseline data for future vector-borne disease monitoring, and control efforts in Kosovo.
Copyright © 2025 Ina Hoxha et al. Transboundary and Emerging Diseases published by John Wiley & Sons Ltd.
Get Full Text
Publication Date: 2025-06-24 PubMed ID: 40599430PubMed Central: PMC12213049DOI: 10.1155/tbed/3208806Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research study details the first recorded instance of West Nile Virus linage 2 (WNV) in mosquitoes observed within the Republic of Kosovo, providing key reference data for further study and control measures for vector-borne diseases in the region.

Research Objectives and Methodology

  • The primary aim of the research was to establish an initial monitoring method for the West Nile Virus (WNV) in the Republic of Kosovo. Kosovo, located in the southern Balkan region, has had limited data on WNV due to an absence of WNV surveillance or specific mosquito monitoring.
  • For this purpose, the researchers performed mosquito sampling in a peri-urban region in the capital city, Prishtina’s western part, from May to September 2022.
  • The collected mosquitoes were pooled together, homogenized, and then their total nucleic acid extracted to perform further tests.

Testing and Result Analysis Process

  • A specific multiplex RT-qPCR, which can identify WNV along with DENV and ZIKV viruses, was utilized. WNV-positive samples were confirmed through a process of RT-PCR and whole-genome sequencing.
  • Out of a total of 44 screened pools, one particular pool, which was molecularly considered to be of the species Culex pipiens sensu largo, tested positive for the presence of WNV RNA.
  • Upon sequencing, this was found to be of the lineage 2 of the WNV. Phylogenetic study further located the sample within a monophyletic clade primarily composed of sequences observed in Southeastern Europe.

Significance of the Study

  • This research is crucial for its evidentiary value – it documents the very first identification of WNV in mosquitoes in Kosovo.
  • Beyond detecting the WNV lineage 2 in Kosovo, the research amasses vital baseline data for informing future actions pertaining to monitoring, analysing, and controlling vector-borne diseases in the region.

Potential Implications

  • Given the geographical spread of the WNV and its impact on humans, this research could help frame effective strategies to preempt or control potential WNV outbreaks in Kosovo and nearby areas.
  • The research may potentially also inform larger-scale global efforts to manage diseases transmitted via vectors like mosquitoes, through its offer of insights into WNV’s regional specificity and transmission patterns.

Cite This Article

APA
Hoxha I, Xhekaj B, Muja-Bajraktari N, Sekulin K, Unterköfler MS, Schlamadinger L, Situmorang T, Fuehrer HP, Obwaller AG, Camp JV, Walochnik J, Sherifi K, Kniha E. (2025). First Detection of West Nile Virus (WNV) Lineage 2 in Mosquitoes in the Republic of Kosovo. Transbound Emerg Dis, 2025, 3208806. https://doi.org/10.1155/tbed/3208806

Publication

Transboundary and emerging diseases
ISSN: 1865-1682
NlmUniqueID: 101319538
Country: Germany
Language: English
Volume: 2025
Pages: 3208806

Researcher Affiliations

Hoxha, Ina
  • Center for Pathophysiology, Infectiology and Immunology, Medical University Vienna, Vienna, Austria.
Xhekaj, Betim
  • Faculty of Agriculture and Veterinary, University of Prishtina, Prishtina, Kosovo.
Muja-Bajraktari, Nesade
  • Faculty of Agriculture and Veterinary, University of Prishtina, Prishtina, Kosovo.
Sekulin, Karin
  • Armaments and Defence Technology Agency, Federal Ministry of Defence, Vienna, Austria.
Unterköfler, Maria S
  • Department of Biological Sciences and Pathobiology, University of Veterinary Medicine Vienna, Vienna, Austria.
Schlamadinger, Lisa
  • Department of Biological Sciences and Pathobiology, University of Veterinary Medicine Vienna, Vienna, Austria.
Situmorang, Tanto
  • Department of Biological Sciences and Pathobiology, University of Veterinary Medicine Vienna, Vienna, Austria.
Fuehrer, Hans-Peter
  • Department of Biological Sciences and Pathobiology, University of Veterinary Medicine Vienna, Vienna, Austria.
Obwaller, Adelheid G
  • Division of Science, Research and Development, Federal Ministry of Defence, Vienna, Austria.
Camp, Jeremy V
  • Center for Virology, Medical University Vienna, Vienna, Austria.
Walochnik, Julia
  • Center for Pathophysiology, Infectiology and Immunology, Medical University Vienna, Vienna, Austria.
Sherifi, Kurtesh
  • Faculty of Agriculture and Veterinary, University of Prishtina, Prishtina, Kosovo.
Kniha, Edwin
  • Center for Pathophysiology, Infectiology and Immunology, Medical University Vienna, Vienna, Austria.

MeSH Terms

  • Animals
  • Kosovo / epidemiology
  • West Nile virus / isolation & purification
  • West Nile virus / genetics
  • West Nile virus / classification
  • Phylogeny
  • Culicidae / virology
  • West Nile Fever / virology
  • West Nile Fever / epidemiology
  • Mosquito Vectors / virology
  • Culex / virology

Conflict of Interest Statement

The authors declare no conflicts of interest.

References

This article includes 47 references
  1. May FJ, Davis CT, Tesh RB, Barrett ADT. Phylogeography of West Nile Virus: From the Cradle of Evolution in Africa to Eurasia, Australia, and the Americas.. Journal of Virology 2011;85(6):2964–2974.
    doi: 10.1128/jvi.01963-10pmc: PMC3067944pubmed: 21159871google scholar: lookup
  2. Hubálek Z, Halouzka J. West Nile Fever–a Reemerging Mosquito-Borne Viral Disease in Europe.. Emerging Infectious Diseases 1999;5(5):643–650.
    doi: 10.3201/eid0505.990505pmc: PMC2627720pubmed: 10511520google scholar: lookup
  3. Bakonyi T, Ivanics É, Erdélyi K. Lineage 1 and 2 Strains of Encephalitic West Nile Virus, Central Europe.. Emerging Infectious Diseases 2006;12(4):618–623.
    doi: 10.3201/eid1204.051379pmc: PMC3294705pubmed: 16704810google scholar: lookup
  4. Bakonyi T, Ferenczi E, Erdélyi K. Explosive Spread of a Neuroinvasive Lineage 2 West Nile Virus in Central Europe, 2008/2009.. Veterinary Microbiology 2013;165(1-2):61–70.
    doi: 10.1016/j.vetmic.2013.03.005pubmed: 23570864google scholar: lookup
  5. World Health Organization. West Nile Virus.. 2024.
  6. Gossner CM, Marrama L, Carson M. West Nile Virus Surveillance in Europe: Moving Towards an Integrated Animal-Human-Vector Approach.. Eurosurveillance 2017;22(18):1–10.
    doi: 10.2807/1560-7917.ES.2017.22.18.30526pmc: PMC5434877pubmed: 28494844google scholar: lookup
  7. Camp JV, Nowotny N. The Knowns and Unknowns of West Nile Virus in Europe: What did we Learn From the 2018 Outbreak?. Expert Review of Anti-infective Therapy 2020;18(2):145–154.
    doi: 10.1080/14787210.2020.1713751pubmed: 31914833google scholar: lookup
  8. Ruscher C, Patzina-Mehling C, Melchert J. Ecological and Clinical Evidence of the Establishment of West Nile Virus in a Large Urban Area in Europe, Berlin, Germany, 2021 to 2022.. Eurosurveillance 2023;28(48).
    doi: 10.2807/1560-7917.ES.2023.28.48.2300258pmc: PMC10690859pubmed: 38037727google scholar: lookup
  9. Pem-Novosel I, Vilibic-Cavlek T, Gjenero-Margan I. First Outbreak of West Nile Virus Neuroinvasive Disease in Humans, Croatia, 2012.. Vector-Borne and Zoonotic Diseases 2014;14(1):82–84.
    doi: 10.1089/vbz.2012.1295pmc: PMC3880908pubmed: 24283515google scholar: lookup
  10. Ahmetagić S, Petković J, Hukić M, Smriko-Nuhanović A, Piljić D. Human West Nile Virus Infection in Bosnia and Herzegovina.. Medicinski Glasnik 2015;12:47–51.
    pubmed: 25669336
  11. Popović N, Milošević B, Urošević A. Outbreak of West Nile Virus Infection Among Humans in Serbia, August to October 2012.. Eurosurveillance 2013;18(43).
    doi: 10.2807/1560-7917.ES2013.18.43.20613pubmed: 24176618google scholar: lookup
  12. Berxholi K, Ziegler U, Rexhepi A. Indigenous West Nile Virus Infections in Horses in Albania.. Transboundary and Emerging Diseases 2013;60:45–50.
    doi: 10.1111/tbed.12141pubmed: 24589101google scholar: lookup
  13. Pervanidou D, Kefaloudi CN, Vakali A. The 2022 West Nile Virus Season in Greece; A Quite Intense Season.. Viruses 2023;15(7).
    doi: 10.3390/v15071481pmc: PMC10383024pubmed: 37515168google scholar: lookup
  14. Dreshaj S, Jakupi X, Sherifi K. Human West Nile Virus Infection in Kosovo 2012–2015: A Cross-Sectional Analysis.. Journal of Neurology & Neurophysiology 2018;960.
  15. European Center for Disease Prevention and Control. Historical Data by Year—West Nile Fever Seasonal Surveillance.. 2019.
  16. Emmerich P, Jakupi X, Sherifi K. Serologic and Genomic Investigation of West Nile Virus in Kosovo.. Viruses 2024;16(1).
    doi: 10.3390/v16010066pmc: PMC10818488pubmed: 38257766google scholar: lookup
  17. Rexhepi A, Sherifi K, Berxholi K. First Serological Evidence of West Nile Virus Among Equines and Birds in Kosovo, 2018–2019.. Vector-Borne and Zoonotic Diseases 2021;21(2):116–120.
    doi: 10.1089/vbz.2020.2673pubmed: 33090084google scholar: lookup
  18. Muja-Bajraktari N, Zhushi-Etemi F, Dikolli-Velo E, Kadriaj P, Gunay F. The Composition, Diversity, and Distribution of Mosquito Fauna (Diptera: Culicidae) in Kosovo.. Journal of Vector Ecology 2019;44(1):94–104.
    doi: 10.1111/jvec.12333pubmed: 31124243google scholar: lookup
  19. Becker N, Petrić D, Zgomba M. Mosquitoes: Identification, Ecology and Control.. 3rd. Cham: Springer International Publishing; 2020.
    doi: 10.1007/978-3-030-11623-1google scholar: lookup
  20. Kolodziejek J, Marinov M, Kiss BJ, Alexe V, Nowotny N, Coffey LL. The Complete Sequence of a West Nile Virus Lineage 2 Strain Detected in a Hyalomma Marginatum Marginatum Tick Collected From a Song Thrush (Turdus philomelos) in Eastern Romania in 2013 Revealed Closest Genetic Relationship to Strain Volgograd 2007.. PLoS ONE 2014;9(10).
    doi: 10.1371/journal.pone.0109905pmc: PMC4184904pubmed: 25279973google scholar: lookup
  21. Pang Z, Li A, Li J. Comprehensive Multiplex One-Step Real-Time TaqMan qRT-PCR Assays for Detection and Quantification of Hemorrhagic Fever Viruses.. PLoS ONE 2014;9(4).
    doi: 10.1371/journal.pone.0095635pmc: PMC3994070pubmed: 24752452google scholar: lookup
  22. Fournier F, Durand F, Estramon E. Real Time Micro-Organisms PCR in 82 Horseflies in France.. Archives of Microbiology & Immunology 2023;7(3):83–95.
    doi: 10.26502/ami.936500107google scholar: lookup
  23. Weissenböck H, Kolodziejek J, Url A, Lussy H, Rebel-Bauder B, Nowotny N. Emergence of Usutu virus, an African Mosquito-Borne Flavivirus of the Japanese Encephalitis Virus Group, Central Europe.. Emerging Infectious Diseases 2002;8(7):652–656.
    doi: 10.3201/eid0807.020094pmc: PMC2730324pubmed: 12095429google scholar: lookup
  24. Smith JL, Fonseca DM. Rapid Assays for Identification of Members of the Culex (Culex) Pipiens Complex, Their Hybrids, and Other Sibling Species (Diptera: Culicidae). American Journal of Tropical Medicine and Hygiene 2004;70(4):339–345.
    doi: 10.4269/ajtmh.2004.70.339pubmed: 15100444google scholar: lookup
  25. Bahnck CM, Fonseca DM. Rapid Assay to Identfy the two Genetic Forms of Culex (Culex) Pipiens L. (Diptera: Culicidae) and Hybrid Populations.. American Journal of Tropical Medicine and Hygiene 2006;75(2):251–255.
    doi: 10.4269/ajtmh.2006.75.2.0750251pubmed: 16896127google scholar: lookup
  26. Young JJ, Haussig JM, Aberle SW. Epidemiology of Human West Nile Virus Infections in the European Union and European Union Enlargement Countries, 2010 to 2018.. Eurosurveillance 2021;26(19).
    doi: 10.2807/1560-7917.ES.2021.26.19.2001095pmc: PMC8120798pubmed: 33988124google scholar: lookup
  27. European Centre for Disease Prevention and Control. West Nile Virus Infection—Annual Epidemiological Report for 2019.. Stockholm, Sweden: 2021.
  28. Lu L, Zhang F, Munnink BBO. West Nile Virus Spread in Europe: Phylogeographic Pattern Analysis and Key Drivers.. PLOS Pathogens 2024;20(1).
    doi: 10.1371/journal.ppat.1011880pmc: PMC10810478pubmed: 38271294google scholar: lookup
  29. Pervanidou D, Vakali A, Georgakopoulou T. West Nile Virus in Humans, Greece, 2018: The Largest Seasonal Number of Cases, 9 Years After its Emergence in the Country.. Eurosurveillance 2020;25(32).
    doi: 10.2807/1560-7917.ES.2020.25.32.1900543pmc: PMC7427301pubmed: 32794446google scholar: lookup
  30. Petrović T, Šekler M, Petrić D. Intensive West Nile Virus Circulation in Serbia in 2018—Results of Integrated Surveillance Program.. Pathogens 2021;10(10).
    doi: 10.3390/pathogens10101294pmc: PMC8540029pubmed: 34684243google scholar: lookup
  31. Vogels CBF, Fros JJ, Göertz GP, Pijlman GP, Koenraadt CJM. Vector Competence of Northern European Culex pipiens Biotypes and Hybrids for West Nile Virus is Differentially Affected by Temperature.. Parasites & Vectors 2016;9(1).
    doi: 10.1186/s13071-016-1677-0pmc: PMC4937539pubmed: 27388451google scholar: lookup
  32. Leggewie M, Badusche M, Rudolf M. Culex pipiens and Culex torrentium Populations From Central Europe are Susceptible to West Nile Virus Infection.. One Health 2016;2:88–94.
    doi: 10.1016/j.onehlt.2016.04.001pmc: PMC5462652pubmed: 28616480google scholar: lookup
  33. Jansen S, Heitmann A, Lühken R. Culex torrentium: A Potent Vector for the Transmission of West Nile Virus in Central Europe.. Viruses 2019;11(6).
    doi: 10.3390/v11060492pmc: PMC6630772pubmed: 31146418google scholar: lookup
  34. Gomes B, Sousa CA, Vicente JL. Feeding Patterns of Molestus and Pipiens Forms of Culex pipiens (Diptera: Culicidae) in a Region of High Hybridization.. Parasites & Vectors 2013;6(1).
    doi: 10.1186/1756-3305-6-93pmc: PMC3637809pubmed: 23578139google scholar: lookup
  35. la Puente JM, Ferraguti M, Ruiz S, Roiz D, Soriguer RC, Figuerola J. Culex pipiens Forms and Urbanization: Effects on Blood Feeding Sources and Transmission of Avian Plasmodium.. Malaria Journal 2016;15(1).
    doi: 10.1186/s12936-016-1643-5pmc: PMC5146868pubmed: 27931226google scholar: lookup
  36. Kothera L, Mutebi J-P, Kenney JL. Host Selection, and Genetic Admixture Analyses of Culex pipiens Complex (Diptera: Culicidae) Mosquitoes in Chicago, IL.. Journal of Medical Entomology 2020;57(1):78–87.
    doi: 10.1093/jme/tjz158pmc: PMC11313203pubmed: 31576405google scholar: lookup
  37. Wilke ABB, Chase C, Vasquez C. Urbanization Creates Diverse Aquatic Habitats for Immature Mosquitoes in Urban Areas.. Scientific Reports 2019;9(1).
    doi: 10.1038/s41598-019-51787-5pmc: PMC6814835pubmed: 31653914google scholar: lookup
  38. Kniha E, Obwaller AG, Dobler G, Poeppl W, Mooseder G, Walochnik J. Phlebovirus Seroprevalence in Austrian Army Personnel Returning From Missions Abroad.. Parasites & Vectors 2019;12(1).
    doi: 10.1186/s13071-019-3674-6pmc: PMC6708154pubmed: 31445517google scholar: lookup
  39. Šolaja S, Goletić Š, Veljović L, Glišić D. Complex Patterns of WNV Evolution: A Focus on the Western Balkans and Central Europe.. Frontiers in Veterinary Science 2024;11.
    doi: 10.3389/fvets.2024.1494746pmc: PMC11614783pubmed: 39634759google scholar: lookup
  40. Chaintoutis SC, Papa A, Pervanidou D, Dovas CI. Evolutionary Dynamics of Lineage 2 West Nile Virus in Europe, 2004–2018: Phylogeny, Selection Pressure and Phylogeography.. Molecular Phylogenetics and Evolution 2019;141.
    doi: 10.1016/j.ympev.2019.106617pubmed: 31521822google scholar: lookup
  41. Fiacre L, Pagès N, Albina E, Richardson J, Lecollinet S, Gonzalez G. Molecular Determinants of West Nile Virus Virulence and Pathogenesis in Vertebrate and Invertebrate Hosts.. International Journal of Molecular Sciences 2020;21(23).
    doi: 10.3390/ijms21239117pmc: PMC7731113pubmed: 33266206google scholar: lookup
  42. Visser I, Marshall EM, Agliani G. In Vitro and in Vivo Characterization of a Novel West Nile Virus Lineage 2 Strain.. npj Viruses 2024;2(1).
    doi: 10.1038/s44298-024-00070-0pmc: PMC11721649pubmed: 40295818google scholar: lookup
  43. Venter M, Myers TG, Wilson MA. Gene Expression in Mice Infected With West Nile Virus Strains of Different Neurovirulence.. Virology 2005;342(1):119–140.
    doi: 10.1016/j.virol.2005.07.013pubmed: 16125213google scholar: lookup
  44. Fall G, Di Paola N, Faye M. Biological and Phylogenetic Characteristics of West African Lineages of West Nile Virus.. PLOS Neglected Tropical Diseases 2017;11(11).
    doi: 10.1371/journal.pntd.0006078pmc: PMC5695850pubmed: 29117195google scholar: lookup
  45. Botha EM, Markotter W, Wolfaardt M. Genetic Determinants of Virulence in Pathogenic Lineage 2 West Nile Virus Strains, Emerg.. Emerging Infectious Diseases 2008;14(2):222–230.
    doi: 10.3201/eid1402.070457pmc: PMC2600181pubmed: 18258114google scholar: lookup
  46. Pérez-Ramírez E, Llorente F, del Amo J. Pathogenicity Evaluation of Twelve West Nile Virus Strains Belonging to Four Lineages From Five Continents in a Mouse Model: Discrimination Between Three Pathogenicity Categories.. Journal of General Virology 2017;98(4):662–670.
    doi: 10.1099/jgv.0.000743pubmed: 28475031google scholar: lookup
  47. Srihi H, Chatti N, Ben Mhadheb M, Gharbi J, Abid N. Phylodynamic and Phylogeographic Analysis of the Complete Genome of the West Nile Virus Lineage 2 (WNV-2) in the Mediterranean Basin.. BMC Ecology and Evolution 2021;21(1).
    doi: 10.1186/s12862-021-01902-wpmc: PMC8477494pubmed: 34579648google scholar: lookup
Subscribe to our newsletter
Join 99,357 horse owners like you who receive our email updates on horse health and nutrition.