From 16 June to 30 July, 2021, a Virginia Tech team of 4-6 sampled the fish community in 30 Shenandoah Valley streams (Virginia and West Virginia, USA) as part of a larger stream-health study including other teams who surveyed geomorphology, water quality, flow, temperature, macroinvertebrates, and fish health at the same 30 streams. The fish community team also sampled three additional bonus streams (SiteID's FOLL, NAKE, and TUMB) during 19-22 August, 2021. These 33 streams had upstream drainage areas of 7 to 46 sq. km and included 4 James River tributaries (SiteID's POAG, MLRB, CEDR, PISG), 1 Potomac River tributary (HARL), and 28 Shenandoah River tributaries. At each stream, we sampled fish from two reaches using two-pass backpack electrofishing and seining. Reach A was the main reach surveyed by all the interdisciplinary teams; Reach B was surveyed for fish communities only. Reach length was 20 wetted channel widths, capped at 150 meters. Fish were identified to species, assigned to a length class, inspected for anomalies, and released. Other measurements included water temperature, electrical conductivity, and clarity, and percent of the reach comprising pool, riffle, and run habitats.