Filarial susceptibility and effects of Wolbachia in Aedes pseudoscutellaris mosquitoes.

The mosquito Aedes pseudoscutellaris (Theobald), a member of the Aedes (Stegomyia) scutellaris complex (Diptera: Culicidae), is an important vector of subperiodic Wuchereria bancrofti (Cobbold) (Spirurida: Onchocercidae), causing human lymphatic filariasis, on South Pacific islands. Maternal inheritance of filarial susceptibility in the complex has previously been asserted, and larval tetracycline treatment reduced susceptibility; the maternally inherited Wolbachia in these mosquitoes were suggested to be responsible. To investigate the relationship of these two factors, we eliminated Wolbachia from a strain of Ae. pseudoscutellaris by tetracycline treatment, and tested filarial susceptibility of the adult female mosquitoes using Brugia pahangi (Edeson & Buckley). Filarial susceptibility was not significantly different in Wolbachia-free and infected lines of Ae. pseudoscutellaris, suggesting that the Wolbachia in these mosquitoes do not influence vector competence. Crosses between Wolbachia-infected males and uninfected females of Ae. pseudoscutellaris showed cytoplasmic incompatibility (CI), i.e. no eggs hatched, unaffected by larval crowding or restricted nutrient availability, whereas these factors are known to affect CI in Drosophila simulans. Reciprocal crosses between Ae. pseudoscutellaris and Ae. katherinensis Woodhill produced no progeny, even when both parents were Wolbachia-free, suggesting that nuclear factors are responsible for this interspecific sterility.

Population differentiation and Wolbachia phylogeny in mosquitoes of the Aedes scutellaris group.

Mosquito species of the Aedes (Stegomyia) scutellaris (Walker) group (Diptera: Culicidae) are distributed across many islands of the South Pacific and include major regional vectors of filariasis, such as Aedes polynesiensis (Marks). Analysis of populations of Ae. polynesiensis at the extremes of its range, from Fiji and from Moorea, French Polynesia, using the rDNA ITS2 (internal transcribed spacer 2) region and six microsatellite markers showed considerable genetic differentiation between them (F(ST) = 0.298-0.357). Phylogenetic analysis of the Wolbachia endosymbionts in three members of the complex revealed that based on the wsp gene they are all very similar and belong to the Mel subgroup of the A clade, closely related to the Wolbachia strain present in the gall wasp Callyrhytis glandium (Giraud) (Hymenoptera: Cynipidae). By contrast they are only distantly related to the A-clade Wolbachia in Aedes albopictus (Skuse), a species closely allied to the Ae. scutellaris group. There was very low differentiation between the Wolbachia in the Moorea and Fiji populations of Ae. polynesiensis.

Strain-specific quantification of Wolbachia density in Aedes albopictus and effects of larval rearing conditions.

The density of the endosymbiont Wolbachia can influence the expression of the crossing sterilities known as cytoplasmic incompatibility (CI), and also its rate of maternal transmission. Aedes albopictus mosquitoes contain a superinfection with the Wolbachia strains wAlbA and wAlbB. A strain-specific real-time quantitative PCR assay was developed and used to quantify relative Wolbachia strain densities within individual mosquitoes. The wAlbB strain was consistently found to be at higher density than wAlbA, which can explain a slightly lower rate of maternal transmission reported for wAlbA. The effects of larval crowding and nutritional stress were also examined. Larval crowding always reduced adult size, but reduced the density of Wolbachia strains relative to uncrowded conditions only if crowding was accompanied by restricted nutrient availability. Crowded rearing conditions never resulted in strain segregation or in a reduction in the penetrance of CI, however. The rate of maternal transmission and the penetrance of CI are the two most important variables that determine relative Wolbachia population invasion dynamics, and both are considerably higher here than have been reported in the Drosophila simulans model system.