Abstract: We evaluated the impact of land cover on fish assemblages by examining relationships between stream hydrology, physicochemistry, and instream habitat and their association with fish responses in streams draining 18 watersheds of the Lower Piedmont of western Georgia. Several important relationships between land use and physicochemical, hydrological, and habitat parameters were observed, particularly higher frequency of spate flows, water temperatures, and lower dissolved oxygen (DO) with percentage impervious surface (IS) cover, higher habitat quality with percentage forest cover, and elevated suspended solid concentrations with percentage pasture cover. Fish assemblages were largely explained by physicochemical and hydrological rather than habitat variables. Specifically, fish species diversity, richness, and biotic integrity were lower in streams that received high frequency of spate flows. Also, overall fish assemblage structure as determined by nonmetric multidimensional scaling was best described by total dissolved solids (TDS) and DO, with high TDS and low DO streams containing sunfish-based assemblages and low TDS and high DO streams containing minnow-based assemblages. Our results suggest that altered hydrological and physicochemical conditions, induced largely by IS, may be a strong determinant of fish assemblage structure in these lowland streams and allow for a more mechanistic understanding of how land use ultimately affects these systems.