The Meramec River Basin in east-central Missouri is an important stronghold for native freshwater mussels (Order: Unionoida) in the United States. Whereas the basin supports more than 40 mussel species, previous studies indicate that the abundance and distribution of most species are declining. Therefore, resource managers have identified the need to prioritize threats to native mussel populations in the basin and to design a mussel monitoring program. The objective of this study was to identify threats of habitat and water-quality degradation to mussel diversity in the basin. Affected habitat parameters considered as the main threats to mussel conservation included excess sedimentation, altered stream geomorphology and flow, effects on riparian vegetation and condition, impoundments, and invasive non-native species. Evaluating water-quality parameters for conserving mussels was a main focus of this study. Mussel toxicity data for chemical contaminants were compared to national water quality criteria (NWQC) and Missouri water quality standards (MWQS). However, NWQC and MWQS have not been developed for many chemical contaminants and some MWQS may not be protective of native mussel populations. Toxicity data indicated that mussels are sensitive to ammonia, copper, temperature, certain pesticides, pharmaceuticals, and personal care products; these compounds were identified as the priority water-quality parameters for mussel conservation in the basin. Measures to conserve mussel diversity in the basin include expanding the species and life stages of mussels and the list of chemical contaminants that have been assessed, establishing a long term mussel monitoring program that measures physical and chemical parameters of high priority, conducting landscape scale modeling to predict mussel distributions, determining sublethal effects of primary contaminants of concern, deriving risk-based guidance values for mussel conservation, and assessing the effects of wastewater treatment plants and non-point source pollution on mussels. A critical next step to further prioritize these needs is to conduct a watershed risk assessment using local data (for example, land use, flow) when available.