Diflubenzuron is a chitin-inhibiting insecticide that is registered for use in controlling a multitude of invertebrate pests. The selective mode of action of diflubenzuron provided us with a unique tool for examining the indirect responses of an experimental aquatic ecosystem following direct impacts at the primary consumer (invertebrate) trophic level. Twelve outdoor aquatic mesocosms (0.1 ha each) were randomly assigned to three treatments of diflubenzuron (four mesocosms/treatment): control, monthly (five total 10-μg/L applications), or biweekly (twice monthly; nine total 10-μg/L applications). Direct impacts on invertebrates (numbers and species composition of insects and zooplankton) and indirect impacts on fish (mortality, growth, and reproduction of bluegill sunfish and largemouth bass), phytoplankton (biomass and productivity), and water quality were measured over 16 weeks posttreatment. Both monthly and biweekly treatments of diflubenzuron decreased total numbers (fivefold reduction) and species richness (twofold reduction) and increased dominance (twofold increase) of zooplankton within 4 weeks posttreatment; Cladocerans, Copepods, and Rotifers were all sensitive. Diflubenzuron reduced species richness of emergent insects, which resulted in increased dominance by a few species; however, impacts on emergent insects were confounded by a treatment × time interaction. Direct reductions in invertebrate grazers caused indirect increases in algal biomass. Indirect effects on biomass (50% reduction) and individual weight (50% reduction) of juvenile bluegill occurred because of apparent decreases in invertebrate food resources. No statistically significant impacts were observed on adult bluegill or largemouth bass for the duration of the experiment. Results indicated that diflubenzuron had both direct and indirect impacts on the experimental aquatic ecosystems under the conditions tested.