A simple and affordable membrane-feeding method for Aedes aegpyti and Anopheles minimus (Diptera: Culicidae).

This study developed an artificial feeding (AF) method to replace direct host feeding (DHF) for the maintenance of Aedes aegypti and Anopheles minimus mosquito colonies. The procedure can be adopted by all laboratories due to its simple and affordable materials and design. The apparatus consists of heparinized cow blood contained in a 5cm diameter glass petri dish with 5cm(2) Parafilm M (Bemis(®)) stretched thinly over the top, with a pre-heated bag of vegetable oil placed underneath to keep the blood warm. Both parts are contained within an insulated Styrofoam™ box with a hole in the lid for mosquitoes to access the membrane. Mosquitoes are fed by AF for 15min at a time. Feeding rate and fecundity of Ae. aegypti mosquitoes feeding on the AF device were compared to those feeding on a live rat (DHF(r)), and of Anopheles minimus mosquitoes feeding on the AF device compared to those feeding on a human arm (DHF(h)). Aedes aegypti mosquitoes fed by AF or DHF(r) had similar feeding rates (38.2±21.5% and 35.7±18.2%, respectively) and overall egg production (1.5% difference). Anopheles minimus mosquitoes fed by the AF method had a lower feeding rate (52.0±1.0% for AF compared to 70.7±20.2% for DHF(h)) and overall egg production (40% reduction compared to DHF(h)). However, the number of eggs produced by AF-fed mosquitoes (1808 eggs per 100 mosquitoes) was still sufficient for colony maintenance, and with increased feeding time both parameters are expected to increase. Reduced feeding rate and overall egg production was observed when Ae. aegypti mosquitoes were fed on blood refrigerated for over two weeks. In conclusion, an AF device has been developed which can replace DHF for Ae. aegypti and An. minimus colony maintenance when using blood refrigerated for a maximum of two weeks.