Appalachian coal surface mines fracture geologic materials, causing release of both major ions and trace elements to water via accelerated weathering. When elevated above natural background, trace elements in streams may produce adverse effects to biota via direct exposure from water and sediment and via dietary exposure in food sources. Other studies have found elevated water concentrations of multiple trace elements in Appalachia's mining‐influenced streams. Excepting Se, trace‐element concentrations in abiotic and biotic media of Appalachian mining‐influenced streams are less well‐known. We analyzed environmental media of headwater streams receiving alkaline waters from Appalachian coal mines for eight trace elements (Al, As, Cd, Cu, Ni, Sr, V, and Zn) and assessed potential consequent ecological risks. Streamwater, particulate media (sediment, biofilm, leaf detritus) and benthic macroinvertebrates (primary consumers, secondary consumers, crayfish) were sampled from six mining‐influenced and three reference streams during low‐flow conditions in two seasons. Dissolved Cu, Ni, and Sr were higher in mining‐influenced streams than in reference streams; whereas Ni, Sr, and Zn in fine sediments and Ni in macroinvertebrates were also elevated relative to reference‐stream levels in samples from mining‐influenced streams. Seasonal ratios of mining‐influenced stream concentrations to maximum concentrations in reference streams also demonstrated mining‐influenced increases for several elements in multiple media. In most media, concentrations of several elements including Ni were correlated positively. All water‐column dissolved concentrations were below protective levels, but fine‐sediment concentrations of Ni approached or exceeded threshold‐effect concentrations in several streams. Further study is warranted for several elements (Cd, Ni, and Zn in biofilms, and V in macroinvertebrates), which approached or exceeded previously established dietary‐risk levels.