Multigenerational toxicological effects of a binary mixture of silver nanoparticles (AgNPs) with glyphosate were identified in Daphnia magna using acute and chronic toxicity tests. Acute toxicity interactions were analyzed with the Abbott method. In the chronic tests, the survival, growth, reproduction, and age at first brood were evaluated for the parents and the exposed (F1E) and non-exposed (F1NE) descendants. The scales tested for binary mixture, at the acute level, presented antagonistic and additive interactions, possibly associated with the complexation of the AgNPs by glyphosate. Multigenerational chronic effects related to the parameters, reproduction, and age at first brood were observed in the descendants tested with the individual compounds, with no recovery for F1E and F1NE. In organisms exposed to binary mixture, there was a delay in the age at first brood and also a significant change in the reproduction parameter, with a strong reduction for the parents, F1E, and F1NE, indicating a higher toxicity than the compounds tested individually. Although the results for acute interactions between AgNP and glyphosate did not provide clear evidence, multigenerational chronic binary mixture trials have resulted in unexpected toxicity compared with individual treatments, increasing the concerns associated with this co-exposure in other scenarios. Therefore, the interaction of binary mixture with the organisms merits further investigation and the results reported in the present study will be useful in this regard.