In 2013, highly virulent West Nile virus (WNV) lineage 2 was repeatedly reported in mosquitoes in South Moravia. This virus is an etiologic agent of West Nile fever, which might lead to meningitis or meningoencephalitis. Unfortunately, no antiviral treatment or vaccine are available at present. In 2018, first neuroinvasive human cases due to WNV, including one fatal case, were documented. These findings prompted the Czech Academy of Sciences and the Regional Authority of the South Moravian Region to establish a monitoring project, aiming at the early detection of WNV in mosquitoes in endemic areas of South Moravia. Other aims are establishing risk assesment for WNV and also dissemination of relevant data about possible health hazards caused by mosquitoes and agents they transmit to general public.