Assessment of sperm motility, apoptosis, and viability of Fathead Minnows (Pimephales promelas) exposed to polyfluorinated alkyl substances (PFAS) at Cape Cod, MA in 2024

Adult male Fathead Minnows (Pimephales promelas) were exposed to water containing per- and polyfluoroalkyl compounds (PFAS) derived from aqueous film-forming foam in mobile laboratories on location at the USGS Toxic Substance Hydrology Groundwater Research Site on Cape Cod, MA. In 2024, to investigate potential effects on male reproductive parameters, two groundwater wells, including one reference (REF) and one PFAS-contaminated water from fire-training area (FTA-2), were used as environmental source waters for continuous-flow fish exposures. Fathead Minnows were acclimated for 21 days in REF water, then exposed to different dilution ratios of FTA-2 and REF treatments for the 21 days. REF served as the reference treatment. After acclimation, the fish were moved into 5 different exposure treatment groups: 1) REF, 2) FTA-2, 3) FTA-2:REF at 10 parts to 90 parts by volume, 4) FTA-2:REF at 25 parts to 75 parts, and 5) FTA-2:REF at 50 parts to 50 parts. To assess potential effects on reproductive parameters, sperm motility measures were collected by using computer assisted sperm motion analysis, and potential effects on programmed testicular cell death were measured with flow cytometry by using live cells stained with Annexin and counterstained with propidium iodide. These data are to be included in a larger study measuring multiple endpoints, including water and tissue chemistry, testis histology, and female reproductive endpoints such as fecundity.