The Clinch River of southwestern Virginia and northeastern Tennessee is arguably the most important river for freshwater mussel conservation in the United States. This featured collection presents investigations of mussel population status and habitat quality in the Clinch River. Analyses of historic water- and sediment-quality data suggest that water column ammonia and water column and sediment metals, including Cu and Zn, may have contributed historically to declining densities and extirpations of mussels in the river's Virginia sections. These studies also reveal increasing temporal trends for dissolved solids concentrations throughout much of the river's extent. Current mussel abundance patterns do not correspond spatially with physical habitat quality, but they do correspond with specific conductance, dissolved major ions, and water column metals, suggesting these and/or associated constituents as factors contributing to mussel declines. Mussels are sensitive to metals. Native mussels and hatchery-raised mussels held in cages in situ accumulated metals in their body tissues in river sections where mussels are declining. Organic compound and bed-sediment contaminant analyses did not reveal spatial correspondences with mussel status metrics, although potentially toxic levels were found. Collectively, these studies identify major ions and metals as water- and sediment-quality concerns for mussel conservation in the Clinch River.