Longitudinal discharge and water-quality campaigns (seepage runs) were combined with surface geophysical surveys, hyporheic zone temperature profiling, and watershed-scale hydrological monitoring to evaluate the locations, magnitude, and impact of stream-water losses from the West Creek sub-basin of the West West Branch Schuylkill River into the underground Oak Hill Mine complex that extends beneath the watershed divide. Abandoned mine drainage, containing iron and other contaminants, from the Oak Hill Boreholes to the West Branch Schuylkill River was sustained during low-flow conditions and correlated to streamflow lost through the West Creek streambed. During high-flow conditions, streamflow was transmitted throughout West Creek; however, during low-flow conditions, all streamflow from the perennial headwaters was lost within the 300 to 600 m “upper reach,” where an 1889 mine map indicated steeply dipping coalbeds underlie the channel. During low-flow conditions, the channel within the “intermediate reach,” 700 to 1,650 m downstream, gained groundwater seepage with higher pH and specific conductance than upstream; however, all streamflow 1,650 to 2,050 m downstream was lost to underlying mines. Electrical resistivity and electromagnetic conductivity surveys indicated conductive zones beneath the upper reach, where flow loss occurred, and through the intermediate reach, where gains and losses occurred. Temperature probes at 0.06 to 0.10 m depth within the hyporheic zone of the intermediate reach indicated potential downward fluxes as high as 2.1 × 10−5 m/s. Cumulative streamflow lost from West Creek during seepage runs averaged 53.4 L/s, which equates to 19.3 percent of the daily average discharge of abandoned mine drainage from the Oak Hill Boreholes and a downward flux of 1.70×10−5 m/s across the 2.1 km by 1.5 m West Creek stream-channel area.