The U.S. Geological Survey (USGS), in cooperation with the Northeast Texas Municipal Water District and the Texas Commission on Environmental Quality, did a baseline assessment in 2010-11 of physical characteristics and selected aquatic biota (fish and mussels) collected at the mesohabitat scale for three stream reaches in the Big Cypress Basin in northeastern Texas for which environmental flows have been prescribed. Mesohabitats are visually distinct units of habitat within the stream with unique depth, velocity, slope, substrate, and cover. Mesohabitats in reaches of Big Cypress, Black Cypress, and Little Cypress Bayous were evaluated to gain an understanding of how fish communities and mussel populations varied by habitat. Selected water-quality properties were also measured in isolated pools in Black Cypress and Little Cypress. All of the data were collected in the context of the prescribed environmental flows. The information acquired during the study will support the long-term monitoring of biota in relation to the prescribed environmental flows.