Rearing Anopheles funestus Mosquitoes

Blood fed An. funestus adult females resting indoor were collected in houses between 06 and 12 AM in Tororo District (0°45′ N, 34°5′E) in Eastern Uganda, an area of high malaria transmission [2] (link). The collection was carried out in April and November 2009. Blood-fed and gravid mosquitoes resting inside houses were collected using aspirators and torches and immediately transported to the laboratory of the National Livestock Resources Research Institute based in Tororo. A new method was used to induce the females to lay eggs. Briefly, the collected blood fed females were stored in net covered paper cups or in cages and provided with cotton wool moistened with sucrose. They were maintained for 4 to 5 days to allow them to fully reach the gravid stage and were checked daily for survival. The gravid mosquitoes were then gently individually introduced into 1.5 ml Eppendorf tubes containing a 1square cm filter paper inserted into the bottom of the tube. The filter paper was moistened and excess water removed. The cap of the Eppendorf tube was pierced with 3 holes to allow air into the tube. The tubes were checked daily for the presence of eggs. Females that laid eggs were carefully removed from the tubes and transferred into Eppendorf tubes with silica gel. Eggs were stored at room temperature or at 4°C for up to 2 days and then sent via courier to the Liverpool School of Tropical Medicine (LSTM) where they were allowed to hatch in small cup and later transferred to larvae bowls for rearing. Apart from 20 families that were reared individually, the egg batches were pooled and reared together. Larvae were comparatively reared in mineral (bottled) and distilled water and fed abundantly with TetraminTM baby fish food every day. Water of each larvae bowl was changed every two days to reduce the mortality. The F₁ adults generated were randomly mixed in cages for subsequent experiments.