Environmental factors influence the effectiveness of microbial control agents in mosquito control programs. Four of these factors (water temperature, larval density, sunlight and the effect of associated filter feeders) were studied with Bacillus thuringiensis var. israelensis under laboratory and semifield conditions in Europe using different instars of Aedes vexans, Ae. aegypti and Culex pipiens. Bioassays conducted at a low temperature (5 degrees C) yielded 10-fold higher LC50 and LC90 values compared with those conducted at a high temperature (25 degrees C). The efficacy of B.t.i. decreased in a linear manner with increasing larval density. Sunlight can reduce the effectiveness of B.t.i. by several times. Competition in food intake by filter feeding Daphnia resulted in lower mortality of mosquito larvae after B.t.i. applications.