Gravid mosquito traps are commonly used for both arbovirus surveillance and population surveillance of mosquitoes of the genus Culex. Oviposition substrates, used as baits in these traps, were tested against Culex under laboratory and field conditions. In the laboratory all substrates tested as 1% and 10% dilutions in 2-choice bioassays against female Cx. quinquefasciatus were significantly more effective than well water controls in eliciting oviposition. Strongest responses were to dilutions of dairy effluent, followed by larval water and infusions of alfalfa hay, alfalfa pellets, Bermuda hay, oak leaves, and Typha leaves, with lowest responses to cow manure infusion. In the field, few significant differences in collections were obtained between traps baited with different infusions. Significantly more Cx. quinquefasciatus and Cx. nigripalpus were collected in traps baited with cow manure infusion (highest) compared to alfalfa hay infusion (lowest). Responses of Cx. quinquefasciatus to dairy effluent and infusions of Bermuda hay, oak leaves, and Typha leaves were not significantly different from either cow manure infusion or alfalfa hay infusion. Responses of Cx. nigripalpus were highest to cow manure infusion and equally low to infusions of alfalfa hay and Typha leaves; moderate responses were observed to dairy effluent and infusions of Bermuda hay and oak leaves. Gravid females comprised 66.7–81.9% of the collections for each infusion type, with no significant difference among infusions in the proportion of gravid females collected.