Depleted Dissolved Oxygen Concentrations Impair the Embryonic and Early Larval Development of Eastern Hellbenders (Cryptobranchus alleganiensis).

Depletion of dissolved oxygen (DO; hypoxia) and its influence on sensitive fauna and vulnerable life stages is an emerging concern in lotic freshwater ecosystems. A species particularly vulnerable to hypoxia is the Eastern Hellbender-a large-bodied, fully aquatic salamander which relies on cutaneous respiration and has evolved to thrive in cold, well-oxygenated streams. Adult hellbenders nest in benthic stream microhabitats which are vulnerable to DO depletion caused by loss of riparian forest cover that increases sedimentation and solar radiation. Hellbender populations are experiencing declines characterized by reduced recruitment in areas with low forest cover, but the influence of hypoxia on hellbender embryonic development is unknown. We hypothesized that hellbender embryos are sensitive to hypoxic conditions because of their high degree of habitat specialization. We conducted two experiments where we exposed freshly laid eggs to a range of DO concentrations in the lab (3-10 mg/L). In both experiments we found that hellbender embryos reared in our lowest DO treatments of 3 and 5 mg/L had an average viability 51% and 34% lower than each experiment's 10 mg/L treatment, respectively. In addition, eggs reared in these low DO concentrations had significantly lower hatching success, a higher percentage of premature hatchlings, and produced hatchlings with smaller morphometrics compared to higher DO treatments. In our second experiment, we also continued to track hatchlings at least 14 days post hatching and demonstrated that premature hatchlings had a high probability of dying within weeks of hatching. Our results indicate that hellbender embryos require DO concentrations of > 5 mg/L for proper development, thus placing them on the more sensitive end of the hypoxia tolerance continuum for freshwater taxa. Our experiments confirmed hellbender embryonic sensitivity to environmentally relevant reductions in DO concentrations and identified future research and conservation needs related to the development of hellbender embryos in the field.