The Trinity River flows through the Dallas-Ft. Worth Metroplex in north central Texas where it receives effluents from numerous point sources including seven large regional wastewater treatment facilities. Historically, the Trinity River has been impacted by massive wastewater loadings which often constitute > 80% of the total river discharge during low flow periods. Normally, high mass loadings correspond to the summer months, compounding the effects of a naturally stressful period, characterized by high temperatures and low dissolved oxygen concentrations. Samples from 12 stations were collected quarterly over an 18 month period from the Trinity River and two tributaries. Water samples were analyzed for a variety of water quality variables, including metals, priority pollutants, pesticides, and general water quality parameters. Water samples were also tested for acute and subchronic effects with several test species. Fish were collected at each station and assemblages were characterized using traditional classification techniques and the Index of Biotic Integrity. In addition, sediment samples were assessed for toxic effects which could have adversely impacted fish recruitment and in situ biomonitoring experiments were performed. Quantitative habitat characterization analyses were performed to gain additional information that could possibly explains differences in fish assemblage structure related to habitat variability. Data were analyzed using regression, univariate, multivariate, and descriptive statistical techniques and new approaches for analyzing impact assessment data were discussed. Results indicated that the most substantial impacts on fish assemblages were confined to a segment of the river where a sequence of point sources, in close proximity to each other, were overloading the river's capacity to sufficiently dilute and/or detoxify the effluent. Data also indicated the presence of episodic toxicity from nonpoint sources. In addition, toxic effects in sediment samples and differences in habitat were detected and may have contributed to measured differences among fish assemblages in the Trinity River.