Identification of Chemicals Associated Gambusia affinis (Cyprinodontiformes: Poeciliidae), and Their Effect on Oviposition Behavior of Culex tarsalis (Diptera: Culicidae) in the Laboratory.

The western mosquitofish, Gambusia affinis (Baird & Girard), has been used worldwide for the control of larval mosquitoes for more than 100 yr. We found that the western encephalitis mosquito, Culex tarsalis Coquillett (Diptera: Culicidae), can detect the presence of G. affinis in oviposition sites based on associated chemicals, leading to a decrease in the number of egg rafts laid. Three volatile chemical compounds were identified in the headspace above the water where G. affinis had been held for 24 h. Oviposition bioassays conducted using standards of the volatile compounds identified (dimethyl disulfide [DMDS], dimethyl trisulfide [DMTS], and S-methyl methanethiosulphonate) found that females reduced oviposition only when low concentrations of DMTS were present, but this response was not consistent across all trials and concentrations tested. DMDS, DMTS, and S-methyl methanethiosulphonate are known bacterial metabolic waste products and may be the source of the compounds. Two nonvolatile compounds of interest were found to be present in the Gambusia-exudate water. After tasting Cx. tarsalis were deterred from ovipositing onto Gambusia-treated water from which the bacteria had been removed by filtration, indicating that the kairomone may consist of nonvolatile compound(s). One of the nonvolatile compounds isolated from the Gambusia-treated water has a benzene ring structure similar to that of cholesterol but the structure of the two nonvolatile deterrents remains to be fully characterized. Our research shows that three volatile compounds and two nonvolatile compounds are present in water associated with G. affinis (Poeciliidae: Gambusia) and affect the oviposition behavior of Cx. tarsalis in laboratory bioassays.

Oviposition Behavior of Culex tarsalis (Diptera: Culicidae) Responding to Semiochemicals Associated with the Western Mosquitofish, Gambusia affinis (Cyprinodontiformes: Poecilliidae).

Considerable previous research has focused on predator-associated semiochemicals and how they affect mosquito oviposition behavior. However, most of this work has been done without taking into consideration either the natural density of the predators or how other semiochemicals in aquatic environments might affect the responses of gravid mosquitoes. The influence of mosquitofish density, source water (tap vs pond), presence of freshly laid egg rafts, and removal of a putative source of semiochemicals (bacteria) on oviposition by Culex tarsalis Coquillett (Diptera: Culicidae) was studied in laboratory bioassays. Culex tarsalis females were deterred from laying egg rafts on water that contained semiochemicals associated with Gambusia affinis (Baird & Girard), but this deterrence was not strongly associated with the density of fish used to condition aged tap water. The number of egg rafts laid onto Gambusia-exudate water made with either tap water (density < 1 fish per liter) or pond water was typically ≥ 50% of that onto water that did not house mosquitofish. Gravid mosquitoes tested individually did not reduce oviposition onto Gambusia-exudate water as compared to controls. Likewise, oviposition by females with ablated wings did not differ significantly between Gambusia-exudate water and controls. Oviposition onto filter-sterilized Gambusia-exudate water was reduced relative to unfiltered water, suggesting that semiochemicals deterring egg-laying were still present after bacteria were removed. Taken together, these findings suggest that the responses of gravid Cx. tarsalis to chemicals from habitats containing mosquitofish are complex and the origin of the semiochemicals present in the Gambusia-exudate water needs to be elucidated.

Oviposition of Culex tarsalis (Diptera: Culicidae) Differs on Water Conditioned by Potential Fish and Insect Predators.

The response of egg-laying Culex tarsalis Coquillett (Diptera: Culicidae) to water conditioned by three fish species used for mosquito control and three predatory aquatic insect species was examined in laboratory binary choice experiments. Oviposition by Cx. tarsalis was 72% less on water conditioned with the arroyo chub, Gila orcutti (Eigenmann & Eigenmann) (Cypriniformes: Cyprinidae) relative to control cups containing aged tap water, but no significant difference was found in the numbers of egg rafts laid on water conditioned with the fathead minnow (Pimephales promelas (Rafinesque), Cypriniformes: Cyprinidae) and the control treatment (water aged 24 h). Mosquito oviposition on water conditioned with the predominantly herbivorous/algivorous California Mozambique tilapia hybrid (Oreochromis mossambicus (Peters) × Oreochromis urolepis hornorum L. (Perciformes: Cichlidae)) or predatory insects (nymphs: Sympetrum corruptum (Hagen) (Odonata: Libellulidae); adults: Thermonectus basillaris (Harris) or Cybister fimbriolatus (Say) (Coleoptera: Dytiscidae)) did not differ significantly relative to that onto water aged for 24 h. As compared with water aged 24 h and water conditioned with diving beetles, oviposition by Cx. tarsalis was significantly lower (≥53%) when live predatory diving beetles were present in oviposition cups. Gravid Cx. tarsalis females do not respond equally to putative semiochemicals in water conditioned with the piscine or aquatic insect predators of immature mosquitoes tested here.