Culex quinquefasciatus, Cx. restuans, Cx. pipiens complex, and Cx. nigripalpus were collected as larvae or egg rafts from the southern USA. Adult female mosquitoes were intrathoracically inoculated with ~1,000 plaque-forming units of West Nile virus (WNV) and saliva was collected from them 5 days later. The amount of infectious WNV in the saliva samples was quantified by plaque assay and WNV RNA was detected by reverse transcriptase polymerase chain reaction (RT-PCR). More than 90% of the mosquitoes had either infectious virus or viral RNA in their saliva. The RT-PCR assay detected a greater percent of samples with WNV RNA than the plaque assay detected infectious virus. Pairwise comparisons revealed 6 significant differences between the 7 groups surveyed. The Cx. nigripalpus secreted lower mean amounts of WNV than 3 other groups, and a difference was found between early- and late-season Cx. quinquefasciatus collected in Louisiana.