Chemical toxicity in the environment may be the consequence of exposure of living organisms to multiple substances, with distinct putative effects. Among this mulytiplicity of chemicals that occur in the wild, pharmaceutical drugs and antifoulers are prone to exert toxic effects on non-target organisms. To characterize the toxicity elicited by a compound of this specific class, the present study used standard and biochemical-based tools to quantify the toxic response of the antifouler and antidandruff zinc pyrithione in Daphnia magna and Daphnia longispina. The analised parameters were immobility, reproduction, behavioral alteration (swimming patterns), anti-oxidant defense (catalase activity), metabolism (GSTs activities), and neurotoxicity (ChE activity) after exposure to sublethal concentrations of this drug. Exposure to zinc pyrithione has been shown to have neurotoxic and oxidative effects, with changes in swimming behavior. There were no changes in reproductive traits of exposed individuals, from both species. The obtained data demonstrate that ecologically relevant levels of zinc pyrithione can deleteriously alter critical parameters in two distinct freshwater microcrustacean species, although with distinct toxicity patterns and outcomes.