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In the late summer of 1998, an outbreak of equine encephalomyelitis due to West Nile virus (WNV) occurred in the Tuscany region of central Italy. The disease was detected in 14 race horses from nine localities in four Provinces: Firenze, Lucca, Pisa and Pistoia. The outbreak area included Fucecchio wetlands (1800 ha), the largest inland marsh in Italy, and the adjacent hilly Cerbaie woodlands with farms breeding horses. To detect potential vectors of WNV, entomological surveys of Fucecchio and Cerbaie were undertaken during 1999–2002 by collecting mosquito larvae from breeding sites and adult mosquitoes by several methods of sampling.Among 6023 mosquitoes (Diptera: Culicidae) collected, 11 species were identified: Aedes albopictus (Skuse), Ae. vexans (Meigen), Anopheles atroparvus Van Thiel, An. maculipennis Meigen s.s., An. plumbeus Stephens, Culex impudicus Ficalbi, Cx. pipiens L., Culiseta longiareolata Macquart), Ochlerotatus caspius (Pallas), Oc. detritus (Haliday) and Oc. geniculatus (Olivier). In Fucecchio marshes, Cx. impudicus predominated with seasonal peak densities in spring and autumn: its greatest abundance during early spring coincides with arrival of migratory birds from Africa. In Cerbaie hills, Cx. pipiens predominated with peak population density in late summer. No viruses were isolated from 665 mosquitoes processed. These findings, plus other data on Italian mosquito bionomics, suggest a possible mode of WNV transmission involving the most abundant Culex in the Fucecchio-Cerbaie areas. Culex impudicus, being partly ornithophilic, might transmit WNV from migratory to non-migratory birds during springtime; Cx. pipiens, having a broader host range, would be more likely to transmit WNV from birds to horses and, perhaps, to humans by late summer.