This study combined sedimentation, sediment chemistry and benthic community data from three Oncorhynchus mykiss cage farms in Ontario, along with a laboratory bioassay to examine the effects of aquaculture waste loading. Waste loading rates, as well as sediment TC, TN, TP and metal (Cu, Zn) concentrations were highest beneath the cage and decreased exponentially with distance. During the 21-day bioassay, Tubifex (Oligochaeta, Naididae), Chironomus (Diptera, Chironomidae) and Sphaerium (Bivalvia, Pisidiidae) were subjected to a gradient of waste loading. Survival and growth of Sphaerium simile was highest at intermediate levels of waste loading. Chironomus riparius growth increased with increasing waste addition. Tubifex tubifex growth increased with exposure to fish waste, compared with the control. Until this study, there were no predictions for thresholds of effect at freshwater cage farms in the literature. The proposed threshold of effect on freshwater benthos is 2.0 - 3.0 g C m-2 day-1.