The release of nanomaterials in water reservoirs is hazardous. Very few reports are available on the interaction of different sized nanoparticles with aquatic organisms and aquatic environment. In the present study, silver nanoparticles (AgNPs) having an average particle size of 20.80 ± 2.31 and 40.04 ± 4.72 nm were synthesized using polyvinylpyrrolidone and l-tyrosine. Ecotoxicological effects of AgNPs were evaluated on less explored crustacean species, Moina macrocopa. The 48 h lethal values (48 h LC₅₀) of 20 and 40 nm AgNPs were 0.11 ± 0.02 and 0.12 ± 0.03 mg/L, respectively. Further, a size dependent inhibition of AgNPs on acetyl cholinesterase and digestive enzymes (trypsin, amylase, β-galactosidase) was observed, while that of the antioxidant enzymes (catalase, superoxide dismutase, glutathione-S-transferase) and alkaline phosphatase were enhanced as compared to control group. These results strengthen the potential of enzymes as biomarker in environmental risk assessment of AgNPs. AgNPs accumulated in the gut of M. macrocopa which could not be completely eliminated, thereby resulting in an increased metal body burden. The accumulation of AgNPs of 20 nm was lower than that of 40 nm indicating the influence of size of nanoparticles on uptake and toxicity. AgNPs agglomerated in moderately hard water medium (MHWM) and this agglomeration influenced the exposure the organism thereto. The size of AgNPs influenced the toxicity to M. macrocopa through interplay between uptake, accumulation, aggregation, and excretion in the organism and environment.