Behaviour and fitness of γHCH/dieldrin resistant and susceptible female Anopheles gambiae and An.stephensi mosquitoes in the absence of insecticide

The effects of gamma HCH/dieldrin resistance genes on various fitness components of mosquito larvae and adult females in the absence of insecticide were investigated in backcrossed strains of Anopheles gambiae Giles and An.stephensi Liston. Among larvae, heterozygotes (RS) developed slightly but significantly faster than homozygotes for resistance (RR) or susceptibility (SS). The lifetime fecundity of RR females in population cages was only half to two-thirds that of SS and RS females despite similar longevities; several reasons were identified: RR gravid females were less responsive to oviposition-site stimuli, their spontaneous activity--as measured in an acoustic actograph--was only half that of SS or RS females, and RR females produced fewer eggs per unit bloodmeal. When inseminated females were recorded in LD 12:12, RR were again less active than SS or RS. When the lighting was switched to a regime simulating full-moonlight, the activity pattern of SS and RS changed and they flew for longer periods. In contrast, the activity of RR females was the same in LD 12:12 as in 'moonlight'. In a test simulation of potential predation, RR mosquitoes took to flight least readily. All component tests on adult females therefore point to RR as being the least fit of the three genotypes. The behavioural tests suggest that resistance has raised the response threshold of RR females to diverse stimuli. A possible physiological mechanism underlying RR behaviour is that a change in the cyclodiene receptor on the chloride channels has increased their permeability to chloride ions, causing hyper-inhibition of the nervous system.     

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.     

Activity and mating competitiveness of γHCH/dieldrin resistant and susceptible male and virgin female Anopheles gambiae and An.stephensi mosquitoes, with assessment of an insecticide-rotation strategy

The effects of gamma HCH/dieldrin resistance genes on flight activity and mating competitiveness were investigated in males from backcrossed strains of Anopheles gambiae Giles and An.stephensi Liston. Activity of males and virgin females of both species, as recorded in an acoustic actograph, occurred mainly at dusk (the E peak). The activity pattern of An.gambiae males was not affected by resistance genes; in mating competition and predator avoidance experiments, however, RR males were less successful than RS males which were less successful than SS males. The activity pattern of An.stephensi differed from An.gambiae in that the E peaks of RR males and females in a gradual dusk regime were out of synchrony with those of SS and RS, the E peaks of RR occurring slightly later. Thus, RR males and females tended to mate assortatively in mate competition experiments. When a sudden dusk regime was substituted for the gradual dusk regime, activity of RR An.stephensi became synchronized with SS and RS activity, but in mating competition experiments RR still tended to mate assortatively. Estimates of male competitiveness, together with previously-obtained estimates of female fitness, were included in population genetics models. Computer simulations showed that the frequency of resistance in populations of An.gambiae and An.stephensi should decrease in the absence of insecticide at a rate comparable with known field reversions.