Freshwater mussels (Unionidae) play a vital role in aquatic ecosystem health, yet populations in Kentucky are experiencing severe declines due to pollution, habitat loss, and anthropogenic pressures. While home to one of the richest freshwater mussels’ diversities in North America, more than half of Kentucky’s mussel species are threatened or endangered. Cryopreservation of gametes has become a valuable conservation tool to safeguard genetic resources and support propagation programs. Research to date has largely emphasized Dimethyl Sulfoxide (DMSO), with limited comparative evaluation of alternatives such as glycerol. Some findings from marine bivalves indicate that DMSO outperforms glycerol in motility and fertilization rates (Table 1). Studies highlighted progress in aquatic cryopreservation, but cryoprotectant evaluation in freshwater mussels is limited (27% of studies, Table 1).
This study will systematically assess DMSO and glycerol at concentrations of 5%, 10%, and 15% for sperm cryopreservation in freshwater mussels. CEROS II Analysis System will be used to measure sperm motility and viability. By integrating CEROS II Analysis technology with cryoprotectant evaluation, this research aims to establish an optimized cryopreservation protocol tailored to freshwater mussels. The outcomes will enhance hatchery propagation, preserve genetic diversity, and strengthen long-term recovery strategies for imperiled mussel populations in Kentucky. Results will also provide broader guidance for restoration initiatives across North America where mussel declines threaten freshwater ecosystem resilience.