This study characterized baseline conditions in minimally impacted headwater streams in three major river drainages in southern West Virginia. Benthic macroinvertebrate samples were collected in the summer and fall of 2013 using a Hess sampler from sites in the New River, Guyandotte River, and Tug Fork drainages. The macroinvertebrates were identified to genus level and several biotic indices were calculated, including the WVSCI and GLIMPSS(CF), to assess stream health.