The eastern hellbender (Cryptobranchus alleganiensis alleganiensis) has experienced precipitous population declines throughout its range. Numerous factors are speculated to be involved, but no empirical evidence has been presented for any. We implemented a population-wide health assessment in Indiana, USA, examining both the physical well-being of individuals and the quality of their habitat. Physicochemical parameters were analyzed directly in the field and later in the laboratory, when appropriate. Samples were collected June 2008-October 2008 and June 2009-September 2009 for reproductive analysis, blood screening, and disease prevalence. Of 27 chemicals screened in water samples, three were found in the study site, including atrazine. Atrazine was found at levels reported to cause reproductive problems in other amphibians. Vitellogenin was detected only in females and proved a reliable indicator of sex. Sperm parameters were generally of high quality and similar to other populations. Most plasma parameters were similar between sexes, although there were significant differences in calcium and potassium concentrations. Abnormalities were common, occurring in 68% of individuals. No hemoparasites were found, but amphibian chytrid fungus (Batrachochytrium dendrobatidis) was detected on one individual. Our findings establish a baseline for hematology and water-quality parameters that can be used as a model for evaluating population health throughout the hellbender range.