Freshwater resources in arid and semi-arid regions are in extreme demand, which creates conflicts between needs of humans and aquatic ecosystems. The Rio Grande basin in the southwestern United States and northern Mexico exemplifies this issue, as much of its aquatic biodiversity is in peril as a result of human activities. Unionid mussels have been disproportionately impacted, though the specific factors responsible for their decline remain largely unknown. This is problematic because the Rio Grande basin harbors one federally endangered unionid mussel (Popenaias popeii, Texas Hornshell) plus two other mussel species (Potamilus metnecktayi, Salina Mucket; and Truncilla cognata, Mexican Fawnsfoot), which are also being considered for listing under the U.S. Endangered Species Act. To date, surveys for these species have not corrected for variability in detection so current range estimates may be inaccurate. Using single occupancy-modeling to estimate detection and occupancy at 115 sites along ~800 river kilometers of the Rio Grande in Texas, we found that detection probabilities were relatively high, indicating that our survey design was efficient. In contrast, the estimated occupancy was low, indicating that our focal species were likely rare within the Rio Grande drainage. In general, the predicted occupancy of our focal species was low throughout their respective ranges, indicating possible range declines. A comparison of currently occupied ranges to presumptive ranges underscores this point. The best-approximating models indicated that occupancy was influenced by habitat, water quantity and quality, and proximity to large-scale human activities, such as dams and major urban centers. We also discuss a series of conservation options that may not only improve the long-term prognosis of our focal species but also other aquatic taxa.