Entomological, veterinary, and human surveillance systems for West Nile virus (WNV) infection have been implemented in Italy since the first detection of the virus in 1998. These surveillance activities documented a progressive increase of WNV activity and spread in different regions and the emergence of new WNV lineages and strains. Italy is a paradigmatic example of the complex epidemiology of WNV in Europe, where sporadic cases of WNV infection, clusters, and small outbreaks have been reported in several regions. In addition, different strains of both WNV lineage 1 and lineage 2 have been identified, even co-circulating in the same area.