The Yankee Fork is a large tributary of the Salmon River located in central Idaho, USA, with an extensive history of placer and dredge-mining activities. Concentrations of selenium (Se) and mercury (Hg) in various aquatic trophic levels were measured in the Yankee Fork during 2001 and 2002. Various measurements of fish health were also performed. Sites included four on the mainstem of the Yankee Fork and two off-channel sites in partially reclaimed dredge pools used as rearing habitat for cultured salmonid eggs and fry. Hg concentrations in whole mountain whitefish and shorthead sculpin ranged from 0.28 to 0.56 μg/g dry weight (dw), concentrations that are generally less than those reported to have significant impacts on fish. Biofilm and invertebrates ranged from 0.05 to 0.43 μg Hg/g dw. Se concentrations measured in biota samples from the Yankee Fork were greater than many representative samples collected in the Snake and Columbia watersheds and often exceeded literature-based toxic thresholds. Biofilm and invertebrates ranged from 0.58 to 4.66 μg Se/g dw. Whole fish ranged from 3.92 to 7.10 μg Se/g dw, and gonads ranged from 6.91 to 31.84 μg Se/g dw. Whole-body Se concentrations exceeded reported toxicological thresholds at three of four sites and concentrations in liver samples were mostly greater than concentrations shown to have negative impacts on fish health. Histological examinations performed during this study noted liver abnormalities, especially in shorthead sculpin, a bottom-dwelling species.