Water chemistry, periphyton and seston chlorophyll a (CHLa), and biological community data were collected from 321 sites from 2001 through 2005 to (1) determine statistically and ecologically significant relations among the stressor (total nitrogen, total phosphorus, periphyton and seston CHLa, and turbidity) variables and response (biological community) variables; and, (2) determine the breakpoint of biological community attributes and metrics in response to changes in stressor variables. Because of the typically weak relations among the stressor and response variables, methods were developed to reduce the effects of non-nutrient biological stressors that could mask the effect of nutrients. Stressor variable concentrations ranged from 0.30 to 11.0 milligrams per liter (mg/L) for total nitrogen, 0.025 to 1.33 mg/L for total phosphorus, 2.9 to 768 milligrams per square meter (mg/m²) for periphyton CHLa, and 0.37 to 42 micrograms per liter (µg/L) for seston CHLa. Turbidity, another stressor variable, ranged from 0.8 to 65.4 Nephelometric turbidity units (NTUs). When the nutrient and CHLa data were compared to Dodds’ trophic classifications, 75.0 percent of the values for total nitrogen, 46.6 percent of the values for total phosphorus, 35.8 percent of the values for periphyton CHLa, and 3.5 percent of the values for seston CHLa, were eutrophic. The invertebrate communities were dominated by families considered highly nutrient tolerant, Chironimidae, (41.7 percent relative abundance), Hydropsychidae, (17.3 percent relative abundance), and Baetidae, (10.2 percent relative abundance). Fish communities were dominated by algivores and nutrient-tolerant species, specifically central stonerollers (13.3 percent relative abundance), creek chubs (9.9 percent relative abundance), and bluntnose minnows (9.3 percent relative abundance). Although not the dominant taxa, white sucker, spotted sucker, green sunfish, and bluegill species were correlated (p ‹0.05) with the stressor variables. The median breakpoints ranged from 2.4 to 3.3 mg/L for total nitrogen, from 0.042 to 0.129 mg/L for total phosphorus, from 54 to 68 mg/m² for periphyton CHLa, from 4.5 to 7.5 µg/L for seston CHLa, and from 14.1 to 16.1 NTU for turbidity. The breakpoints determined in this study, in addition to Dodds’ trophic classifications, were used as multiple lines of evidence to show changes in fish and invertebrate community and attributes based on annual exposure to nutrients.