The former Callahan Mine Site in Brooksville, ME is an open-pit, hardrock mine site in an intertidal system, providing a unique opportunity to evaluate how metal-enriched sediments and overlying water impact estuarine food webs. Cu, Zn, Cd, and Pb concentrations in sediment, whole water, and Atlantic killifish (Fundulus heteroclitus) were evaluated at sites in Goose Pond (the Callahan Mine Site) and at reference sites. The metal concentrations of sediment, water, and fish were spatially distinct and significantly higher at the Mine Site than in the reference estuary. Sediment concentrations were particularly elevated and were above probable effects levels (PEL) for all four metals adjacent to the tailings pile. Even in this well-mixed system, water metal concentrations were significantly elevated adjacent to the tailings pile and the concentrations of Cu and Zn were above ambient water quality criteria (AWQC) for chronic marine exposure. Neither organic matter in the sediment nor salinity or pH of the water explained the metal concentrations. Adjacent to the tailings pile, killifish body burdens were elevated and were significantly related to both sediment and aqueous concentrations. In conclusion, (1) the contaminated sediment and seepage from the tailings impoundment and waste rock pile 3 create a continual flux of metals into the water column, (2) the metals are bioavailable and are bioconcentrating as evident in the killifish tissue concentrations, and (3) Callahan Mine is directly affecting metal bioaccumulation in fauna residing in the Goose Pond estuary and, potentially, in Penobscot Bay via the ‘trophic nekton relay.’