We sampled 240 wadeable streams across Wisconsin for different forms of phosphorus and nitrogen, and assemblages of macroinvertebrates and fish to (1) examine how macroinvertebrate and fish measures correlated with the nutrients; (2) quantify relationships between key biological measures and nutrient forms to identify potential threshold levels of nutrients to support nutrient criteria development; and (3) evaluate the importance of nutrients in influencing biological assemblages relative to other physicochemical factors at different spatial scales. Twenty-three of the 35 fish and 18 of the 26 macroinvertebrate measures significantly correlated (P < 0.05) with at least one nutrient measure. Percentages of carnivorous, intolerant, and omnivorous fishes, index of biotic integrity, and salmonid abundance were fish measures correlated with the most nutrient measures and had the highest correlation coefficients. Percentages of Ephemeroptera-Plecoptera-Trichoptera individuals and taxa, Hilsenhoff biotic index, and mean tolerance value were macroinvertebrate measures that most strongly correlated with the most nutrient measures. Selected biological measures showed clear trends toward degradation as concentrations of phosphorus and nitrogen increased, and some measures showed clear thresholds where biological measures changed drastically with small changes in nutrient concentrations. Our selected environmental factors explained 54% of the variation in the fish assemblages. Of this explained variance, 46% was attributed to catchment and instream habitat, 15% to nutrients, 3% to other water quality measures, and 36% to the interactions among all the environmental variables. Selected environmental factors explained 53% of the variation in macroinvertebrate assemblages. Of this explained variance, 42% was attributed to catchment and instream habitat, 22% to nutrients, 5% to other water quality measures, and 32% to the interactions among all the environmental variables. ?? 2006 Springer Science+Business Media, Inc.