Optimal life stage for radiation sterilization of Anopheles males and their fitness for release.

Mating by male Anopheles mosquitoes (Diptera: Culicidae) was evaluated in the laboratory to assess fitness effects of radio-sterilization applied during different life stages of the malaria vectors An. stephensi Liston and An. gambiae Giles sensu stricto. After reproductive sterilization by gamma-irradiation (120 Gy), equal proportions of sterile and fertile (unirradiated) male adults were released into cages with virgin females and allowed to compete for matings. Radio-sterilization was applied when the males were pupae aged 0-7 h or 24-32 h, or adults aged <24 h or 24-55 h. After being radio-sterilized in the adult stage, males of both species competed effectively with unirradiated males, whereas those sterilized in the pupal stage obtained significantly fewer matings than unirradiated males from the same cohort. There was no evidence of females obtaining multiple inseminations. These findings emphasize the need to radio-sterilize males as adults in order to minimize the fitness cost. Such males may be intended for sterile insect technique population suppression or for trial releases of transgenic anophelines.

In situ hybridization to the Rdl locus on polytene chromosome 3L of Anopheles stephensi.

We are interested in generating a Y-autosome translocation of the Resistance to dieldrin (Rdl) locus in the malaria vector mosquito Anopheles stephensi Liston (Diptera: Culicidae), for use in sterile insect release. To ensure stability of the system, a recombination suppressing inversion can also be induced which encompasses the Rdl locus. As a first step, here we report the cloning of fragments of the Rdl gene from both An. stephensi and An. gambiae Giles using degenerate primers in the polymerase chain reaction. These fragments encode the second membrane-spanning region of the gamma-aminobutyric acid receptor and show high levels of both nucleotide and predicted amino acid identity to other Rdl-like receptors. They confirm that, as in all other arthropod species examined, dieldrin resistance in An. stephensi is associated with replacement of alanine302, in this case with a serine. In situ hybridization of the Rdl probe to polytene chromosomes of An. stephensi localizes the gene to the left arm of chromosome 3 (3L) in region 45C. Rdl localization will enable us to identify chromosomal rearrangements encompassing the Rdl locus and help anchor the genome sequence of An. gambiae to the polytene map.