Nanoplastics (NPs; <0.1 μm) are speculated to be a bigger ecological threat due to their predicted wider distribution, higher concentrations, and bioavailability. Primary NPs are manufactured to be that size, while secondary NPs originate from fragmentation of bigger debris. To date, the long-term impact of NPs in freshwater systems, particularly secondary NPs, is not well-understood. Thus, we employed a freshwater invertebrate, Daphnia magna, to investigate the chronic effects of model primary NPs, fluorescent polystyrene nanospheres (PS-NPs; 20 nm), and water leachate of weathered single-use plastics that contained micro- and nanosized particles. In experiment 1, parent Daphnia (F0) were exposed to 1 and 50 mg/L PS-NPs until the production of the neonates (F1) followed by a two-generation recovery. PS-NPs were mainly detected in the intestine and brood chamber in F0 and transferred to F1 and F2. PS-NPs significantly decreased the appendage curling and heartbeat rate in F0 and reduced reproduction in F2. In experiment 2, the plastic leachate also reduced the appendage curling rate but increased growth and reproduction. The results suggest that the acute toxicity of primary and secondary plastic particles is low even at high concentrations, but their chronic and sublethal effects should not be overlooked.