Abstract Exposure of Daphnia pulex to the insecticide and juvenile hormone-mimic methoprene resulted in a decrease in the incidence of all-male broods and an increase in the incidence of all-female broods compared with controls. These effects were observed at nominal concentrations of 10 and 100 μg/L, within the upper range of concentrations at which methoprene is applied in the environment. Because methoprene has been found to bind to the mammalian retinoid × receptor, we also tested the effects of retinoic acid on Daphnia reproduction. Neither 9-cis-retinoic acid nor all-trans-retinoic acid had any observable effect. Because juvenile hormone and ecdysteroids interact in many insect systems, we also exposed Daphnia to 20-OH-ecdysone. Exposure to the crustacean hormone 20-OH-ecdysone at levels of 1 and 10 μg/L resulted in an increase in all-male broods and a decrease in all-female broods, but 100 μg/L 20-OH-ecdysone resulted in a decrease in all-male broods and an increase in all-female broods. Our results suggest that juvenile hormone and ecdysteroids might play a role in the Daphnia sex determination system.