Sandy Run (Vinton County, southeastern Ohio, USA) is a stream receiving acid mine drainage (AMD) from an abandoned coal mine complex. This stream has been dammed to form Lake Hope. The heavy metal composition of waters (benthic and pore), sediments, and macroinvertebrates in the lake reservoir sediments were analyzed. Lake waters contained Mn as the heavy metal present in higher concentrations followed by Fe, Al, and Zn. Depletion of Fe and Al occurred from precipitation of less soluble Fe and Al oxides and hydroxides along Sandy Run before entering the lake, producing a high Mn water input into the reservoir. Concentrations of heavy metals in the sediments increased toward the dam area. Sequential extraction of metals in the sediments showed that the highest fractions of metals corresponded to the detrital fraction or eroded material from the watershed and metals associated with iron and manganese hydroxides. Heavy metals in the organic sediment fraction were low. Heavy metals from the AMD source, as well as sediments rich in heavy metals eroded from the watershed, were transported to the downstream dam area and stored at the bottom, producing the observed chemistry. Heavy metals in benthic waters also were sourced from the diffusion of ions from sediments and lake waters as variation in pH and redox conditions determined the flux at the sediment-water interface. Metal concentrations were measured within two deposit feeders, oligochaetes and chironomids, and compared to trends in physical metal concentration across the lake. For the four heavy metals with higher concentration in both benthic animals, the concentrations followed the trend: Fe>Al>Mn>Zn, which were similar to the bioavailable metals in the sediments rather than the pore or the benthic water where Mn was the most abundant heavy metal. Ingestion of sediment, not exposure to pore or benthic waters, appeared to be the main transfer mechanism for metals into the biota. Trends and patterns in animal metal concentrations across the lake were probably a complex process controlled by metabolic needs and metallic regulation and tolerance. Even when Mn was the highest concentration heavy metal in the pore waters, it was the lowest to bioconcentrate in the organisms. In comparison, Cd, the lowest concentration metal in the sediments, presented one of the highest bioaccumulation factors.