A bunyavirus surveillance was performed in 2,600 pools consisting of 45,728 mosquitoes collected in north-central Florida from May 2006 to April 2007. Fifteen mosquito pools were found to be virus-positive from the total 2,600 mosquito pools tested (0.6% infection rate), which resulted in a minimum infection rate of 0.33 per 1,000 mosquitoes. Sequence data identified the virus to be Tensaw virus, a member of the Bunyaviridae family. All the virus-positive samples were obtained from pools collected from May to October 2006, in 3 of the 4 major locations studied, revealing the presence of Tensaw virus in north-central Florida mosquito populations in 2006.