Anopheles gambiae s.s. Giles accepted a range of agarose gels, varying from 0.5 to 8% (wt:vol), for oviposition; laid more eggs on 0.5% agarose gels than on moist filter paper or on drier agarose; and laid equal numbers of eggs on 0.5% agarose gels and distilled water. Larvae hatched on agarose gel substrates and crawled onto the surface of moist agarose gels, but they tended only to burst the egg cap and remain within the egg case on drier gels. A mixture of cultured bacteria, originating from a natural larval habitat of An. gambaiae s.s. in western Kenya, was classified by 16S rDNA sequence analysis, and then this mixture or individual colonies from it was used as odor sources in ovipositional experiments with agarose substrates. Of 61 sequences from a mixed clone sequence library, most (78%) were Pseudomonas strains, whereas the remainder were Stenotrophomonas, Enterobacter, Pantoea, Klebsiella, Acinetobacter, Aeromonas, and Bacillus. Oviposition was significantly reduced when mixtures of bacterial colonies of these strains, or a lawn of colonies of a field isolate of S. maltophilia, was presented. Oviposition was neither reduced nor enhanced when field isolates of Pseudomonas putida or Pseudomonas alcaligenes colonies were presented. These results suggest that gravid An. gambiae females are sensitive to bacterial-derived odors emanating from cultured bacteria from natural larval habitats and that some bacterial odors may be repellent.