In the Appalachian region, numerous streams are polluted with acid mine drainage (AMD). These waters are sulfate‐rich with elevated amounts of total acidity, low pH, and high levels of dissolved metals. Biotic multimetric indices, such as the Periphyton Index of Biotic Integrity (PIBI) have been employed to determine water quality across a variety of environmental conditions and may prove useful for AMD impacts. This study was initiated (1) to evaluate the PIBI for distinguishing AMD impact in streams and (2) to examine whether PIBI scores are impacted by seasonal differences. Twelve AMD and three reference streams were sampled for periphyton in June, August, and October. Water chemistry was collected at least once during the sampling period. Preliminary results showed that PIBI scores were significantly different (p<0.05) among the seasons. In addition, the seasonal trends in PIBI scores among streams were not consistent. The PIBI scores were correlated with six water chemistry variables in August and with at least one variable indicative of AMD in each season sampled. PCA and UPGMA analyses of water chemistry data grouped the streams into five categories: (1) moderately impacted AMD streams with lower total dissolved solids, sulfate, total aluminum, and alkalinity, and higher sulfate; (2) AMD streams with higher alkalinity and lower total aluminum; (3) AMD streams with lower alkalinity and higher total aluminum; (4) reference streams and (5) an outlier reference stream affected by nutrients. Relationships between the groups based on water chemistry and the groups derived from the PIBI will be discussed.