Factory Creek is a major tributary to Shoal Creek (Tennessee River drainage) and drains 311 km² in Lawrence and Wayne counties, TN. We conducted a fish survey of Factory Creek in 2017 and calculated index of biotic integrity (IBI) scores using the “30 + 2” sampling method of the Geological Survey of Alabama for Tennessee River Valley streams. Our collections yielded 58 fish species, with a mean of 27 species per collection and with Erimyzon claviformis (Western Creek Chubsucker) reported for the first time from Factory Creek. Including historical records, the known fish fauna of Factory Creek comprises 75 species. Index scores varied from 42 to 58 out of 60 (good to excellent) with a mean of 52 (excellent), which reflects a relatively undisturbed and intact fish community. Factory Creek is located in the Pickwick Lake subbasin, a hotspot for aquatic biodiversity in the southeastern United States. Because much of Factory Creek has excellent IBI conditions, we recommend continued biological monitoring and adherence to best land-use management practices to ensure proactive conservation of the fish fauna.