The Alligator Snapping Turtle (Macrochelys temminckii) is a large freshwater turtle endemic to river systems that drain into the Gulf of Mexico. Turtle populations were sharply reduced by commercial harvest in the 1970s and 1980s; however, the species has yet to be protected under the Endangered Species Act. While anthropogenic stressors such as habitat fragmentation and degradation and illegal capture continue to threaten populations, the degree to which disease may be contributing to any decline of the Alligator Snapping Turtle is unknown. Data were collected from 97 free-ranging Alligator Snapping Turtles in nine waterways in Florida and Georgia from 2001 to 2006. Eleven turtles were captured more than once, resulting in a total sample pool of 123. Reference ranges were established for complete blood count, plasma biochemistry values, trace metals (mercury, zinc, copper, lead, and arsenic), and nutrient parameters (vitamins A, E, D, and selenium). Variations by capture location, sex, and season were detected and likely resulted from external factors such as habitat and diet. Turtles sampled in one location were positive for tortoise herpesviral antibodies. Blood mercury values also differed among populations. This study provides justification for the use of these long-lived aquatic turtles as biologic monitors of the health of local freshwater ecosystems.