Animals in many freshwater habitats are experiencing decreased recruitment due to declines in reproductive health. Both subspecies of a long-lived aquatic salamander, (Cryptobranchus alleganiensis alleganiensis and C. a. bishopi) have experienced severe population declines characterized by low recruitment. For many states throughout their geographic range, captive propagation and translocation are the only remaining form of management given the severity of declines. These captive rearing programs should rely on techniques to assess male reproductive health, which are currently lacking. In this study, we compared the sperm health (motility, viability, and concentration) of male hellbenders from declining and stable populations. Sperm motility and viability were similar among populations, but sperm concentrations (sp/ml) were significantly lower in declining Missouri populations than in hellbenders from populations with higher recruitment in the southeast. Sperm from Ozark hellbenders was successfully cryopreserved but with low post thaw motilities. This method for assessing male reproductive health provides the first baseline comparative study among populations of this cryptic species in decline and has broad implications for use in monitoring male health across the distribution of the eastern hellbender.