Use of rDNA-PCR to investigate the ecological distribution of Anopheles bwambae in relation to other members of the An-gambiae complex of mosquitoes in Bwamba County, Uganda

Environmental relationships were investigated among three species of the Anopheles gambiae complex of mosquitoes associated with the geothermal springs located in Bwamba County, Uganda. The degree of ecological isolation between An.gambiae and An.bwambae, a sibling species known only from the geothermal springs environment, was assessed on the basis of adult distribution and abundance as well as differences in larval habitats. Field data were gathered during June 1995 without knowing which of the species were being collected. Specimens identified subsequently by rDNA-PCR were used to interpret the ecological data. Ten of twenty aquatic sites sampled were found positive for immature stages of the An.gambiae complex. Larvae of An.bwambae were associated with 'springwater' habitats having much higher conductivity, much greater concentrations of dissolved solids and slightly higher temperature and pH than 'normal' fresh water sites inhabited by larvae of An.gambiae. Larval habitats of both species were unshaded: An.bwambae occurred among dense sedge (Cyperus laevigatus) whereas those of An.gambiae were almost devoid of vegetation. One mixed sample showed that larvae of both species occur together in peripheral aquatic sites with intermediate physical and ecological characteristics. In water preference tests, free-flying females were reluctant to lay eggs on bowls of water in cages; gravid females (with one wing amputated) placed on the surface of water in a cup laid eggs on seasoned rainwater (12/51 An.bwambae; 2/3 An. gambiae) as well as springwater (39/51 An.bwambae; 1/3 An.gambiae). All three An.gambiae oviposited on the first water option, whereas 86% of An.bwambae witheld oviposition until being moved to the other type of water after 5-6 h, and 82% (36/44) of these laid eggs on geothermal water in preference to rainwater. Larval and adult collections showed that An.gambiae occurs sympatrically with An.bwambae throughout its range in the humid foothill environment of the geothermal springs, whereas the distribution of An, arabiensis overlaps only slightly with An.bwambae towards the savanna environment north of the springs.