Carbendazim is a fungicide largely used in agriculture as a plant protection product. As a result of agricultural runoffs, drainage, and leaching, it reaches surface waters at concentrations possibly hazardous to aquatic communities. Because of potential and continuous release of carbendazim to aquatic systems, long-term exposure to aquatic organisms should be addressed. To fill the knowledge gap, the present study evaluated the responses of multiple generations of Daphnia magna (clone K6) to an environmentally relevant concentration of carbendazim (5 μg/L). Twelve successive generations were evaluated, and the effects in these offspring were compared with those from a control population. Neonates' fitness was assessed through immobilization, reproduction, and feeding activity tests, along with the comet assay for in vivo DNA damage evaluation. Recovery from long-term exposure was also assessed. In the F5 generation, the results revealed that when daphnids were re-exposed to carbendazim, DNA damage was higher in daphnids continuously exposed to carbendazim than those from clean medium. After daphnids were moved to a clean medium, a low recovery potential was observed for DNA damage. Daphnids exposed continuously for 6 generations (F6) to carbendazim displayed an increase in feeding rates when re-exposed to carbendazim compared with F6 daphnids reared in clean medium. Continuous exposure of daphnids to carbendazim induced a significant increase in DNA damage from the F0 to the F12 generation. Deleterious effects of the multigenerational exposure to carbendazim were more prominent at a subcellular level (DNA damage) compared with the individual level. Environ Toxicol Chem 2017;36:383-394. © 2016 SETAC.