The overwintering strategy of the mosquito Culex modestus, an important West Nile virus (WNV) vector in Europe, was explored under field conditions in reedbed (Phragmites australis) ecosystems in early 2019. A total of 30 Cx. modestus females were found in a BG-Sentinel trap placed in a plastic greenhouse as well as in a reference BG-Sentinel trap placed under the open sky, both set up within the reedbeds and inspected every 2-3 days from February 27 to April 10, 2019. Moreover, 186 females of Cx. pipiens, 3 females of Anopheles hyrcanus, and 3 females of Culiseta annulata were trapped in the monitored time span. While all Cx. modestus females tested negative for the presence of WNV and other arboviruses circulating in Central Europe, we confirmed WNV lineage 2 and Ťahyna virus infection in several pools of the collected Cx. pipiens, demonstrating arbovirus overwintering. This pilot study highlights the need for large-scale monitoring activities covering different regions to identify the overwintering strategy of both mosquito-borne viruses and their vectors in Central Europe.

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