Although mercury (Hg) is a well-studied contaminant, knowledge about Hg accumulation in snakes is limited. The authors evaluated Hg bioaccumulation within and among four snake species (northern watersnakes, Nerodia sipedon; queen snakes, Regina septemvittata; common garter snakes, Thamnophis sirtalis; and rat snakes, Elaphe obsoleta [Pantherophis alleghaniensis]) from a contaminated site on the South River (Waynesboro, VA, USA) and two nearby reference sites. Total Hg (THg) concentrations in northern watersnake tail tissue at the contaminated site ranged from 2.25 to 13.84 mg/kg dry weight (mean: 4.85 ± 0.29), or 11 to 19 times higher than reference sites. Blood THg concentrations (0.03-7.04 mg/kg wet wt; mean: 2.24 ± 0.42) were strongly correlated with tail concentrations and were the highest yet reported in a snake species. Within watersnakes, nitrogen stable isotope values indicated ontogenetic trophic shifts that correlated with THg bioaccumulation, suggesting that diet plays a substantial role in Hg exposure. Female watersnakes had higher mean THg concentrations (5.67 ± 0.46 mg/kg) than males (4.93 ± 0.49 mg/kg), but no significant differences between sexes were observed after correcting for body size. Interspecific comparisons identified differences in THg concentrations among snake species, with more aquatic species (watersnakes and queen snakes) accumulating higher mean concentrations (5.60 ± 0.40 and 4.59 ± 0.38 mg/kg in tail tissue, respectively) than the more terrestrial species, garter snakes and rat snakes (1.28 ± 0.32 and 0.26 ± 0.09 mg/kg, respectively). The results of the present study warrant further investigation of potential adverse effects and will aid in prioritizing conservation efforts.