Multiple physical and chemical stressors can simultaneously affect the biological condition of streams. To better understand the complex interactions of land-use practices, water quality, and ecological integrity of streams, the U.S. Geological Survey National Water Quality Assessment Project is conducting regional-scale assessments of stream condition across the United States. In the summer of 2013, weekly water samples were collected from 100 streams in the Midwestern United States. Employing watershed theory, we used structural equation modeling (SEM) to represent a general hypothesis for how 16 variables (previously identified to be important to stream condition) might be inter-related. Again, using SEM, we evaluated the ability of this “stressor network” to explain variations in multimetrics of algal, invertebrate, and fish community health, trimming away any environmental variables not contributing to an explanation of the ecological responses. Seven environmental variables—agricultural and urban land use, sand content of soils, basin area, percent riparian area as forest, channel erosion, and relative bed stability—were found to be important for all three-community metrics. The algal and invertebrate models included water-chemistry variables not included in the fish model. Results suggest that ecological integrity of Midwest streams are affected by both agricultural and urban land uses and by the natural geologic setting, as indicated by the sand content of soils. Chemicals related to crops (pesticides and nutrients) and residential uses (pyrethroids) were found to be more strongly related to ecological integrity than were natural factors (riparian forest, watershed soil character).