West Nile virus transmission in the Metropolitan Area of Barcelona (Spain): A One-Health surveillance approach.

West Nile virus (WNV), mainly transmitted by mosquitoes, poses significant health risks to humans and horses, particularly in endemic regions. The first detection of WNV lineage 2 in Spain was in 2017 in Catalonia (northeastern Spain). In 2023, WNV was confirmed in a young yellow-legged gull and a probable human case was notified within the urban settings. We aimed to define the zone of WNV circulation in the Barcelona Metropolitan Area where these infections occurred and the effectiveness of the One Health approach for early WNV detection. The Catalan WNV surveillance and control programs includes the testing of horses, birds, mosquitoes and humans following molecular and serological methods. Phylogenetic analyses were performed to determine the origin of the circulating virus. IgM-positive data from both active and passive surveillance in horses identified the area of WNV circulation and suggested that WNV circulation happened either before or concurrently with human and bird infections in the agricultural and peri-urban areas. Furthermore, a new WNV introduction was discarded by phylogenetic studies, demonstrating that WNV lineage 2 has been established in Catalonia, albeit at a low level of circulation since the virus was not detected in blood donors. Our findings underscore the importance of integrating active and passive surveillance strategies to early assess WNV circulation and activate public health responses. The study highlights the role of wildlife in the WNV transmission and emphasizes the need for ongoing monitoring in animals and also mosquito control measures to mitigate the risk of animal and human infections.