ORIGINAL RESEARCH article

Front. Microbiol.

Sec. Virology

Detection and genomic characterization of West Nile virus lineage 2 in captive harbor seals in Southern Italy

  • 1. Experimental Zooprophylactic Institute of Southern Italy (IZSM), Portici, Italy

  • 2. Universita Campus Bio-Medico di Roma, Rome, Italy

  • 3. Azienda Sanitaria Locale Napoli 2 Nord, Pozzuoli, Italy

  • 4. Universita degli Studi di Napoli Federico II, Naples, Italy

The final, formatted version of the article will be published soon.

Abstract

West Nile virus (WNV) is a mosquito-borne flavivirus maintained in bird-mosquito cycles, with occasional spillover events causing neuroinvasive disease in humans and animals. This study describes two fatal cases of West Nile virus (WNV) lineage 2 infection in captive harbor seals housed in a zoological park in southern Italy, together with genomic characterization and phylogenetic analysis of the detected strains. Harbour seals were tested by multiplex real-time RT-PCR for the simultaneous detection of lineage 1 and 2. Next, selected extracts were analyzed by droplet digital PCR (dd-PCR). Next-generation sequencing was performed and phylogenetically analyzed. A CDC light trap was used for targeted mosquito collection near the fence of the dead harbor seals and female mosquitoes were screened for WNV. Histopathology was also carried out in one seal. WNV lineage 2 RNA was detected in central nervous system tissues of both seals and in female Culex pipiens pool collected. Near-complete genomes clustered with contemporary southern Italian strains and was consistent with local circulation. These findings document WNV lineage 2 infection associated with fatal neurological disease in harbor seals, expanding the knowledge of host range of infection of this virus, also supported by in-situ entomological and genomic evidence. Altogether, this study highlights the value of integrated One Health surveillance approach in zoological parks

Summary

Keywords

Fatal cases, Genetic characterization, Harbor seals, lineage 2, West Nile virus

Received

14 July 2026

Accepted

03 August 2026

Copyright

© 2026 De Martinis, Cardillo, Giovanetti, Pucciarelli, Picazio, Viscardi, Cozzolino, Cavaliere, Napolitano, Lella, Dimatteo, Silvestre, Di Napoli, D'Alessio, De Carlo, GALIERO, Iovane, Paciello and Fusco. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

*Correspondence: Lorena Cardillo

Disclaimer

All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.

Outline

Share article

Article metrics