Our project sought to determine ecological effects of adding low-head dams and levees to large rivers by examining potential changes to aquatic food webs over a 70-year period in the Lower Ohio River (LOR) and Upper Mississippi River (UMR). We employed museum collections of fish and compound specific stable isotope analysis of amino acids to evaluate long-term changes in primary food sources for multiple species of fish in each river. Fishes in both rivers depended more on autochthonous than allochthonous carbon sources throughout the 70-year period (based on measurements of isotopic signatures of algae, C-3 plants, C-4 plants, cyanobacteria, and fungi), but the relative use of different carbon sources differed between the UMR and LOR. Significant but opposite shifts in trophic positions (TP) between rivers over time (higher TP in the UMR; lower in the LOR) were correlated with major anthropogenic changes to habitat structure (e.g. slight decrease in abundance of side channels in the UMR; increase in pool water depth in the LOR) resulting from low-head dam construction. They may also have been influenced by likely increased primary productivity in the UMR from agricultural nitrogen inputs and by possible shifts in the importance of phytoplankton versus benthic algae in the LOR from changes in water depth. Shifts in trophic position and reliance on various food sources were not correlated with variation in discharge, gage height, or temperature. Although these two rivers have contrasting hydrogeomorphic complexity (UMR is an anastomosing river, while the LOR is a constricted channel river) and different discharge patterns (seasonal versus yearly operation in some cases), both differ substantially from rivers having hydrogeomorphic changes resulting from construction of high dams (>15 m). It is not surprising, therefore, that factors controlling trophic position and reliance on different carbon sources vary among different types of dams and river structures.