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Cypress Creek is a northern tributary to the Tennessee River (Pickwick Lake) in Lauderdale County, Alabama, and Wayne County, Tennessee, with a drainage area of 214 square miles. Although repeatedly sampled for fishes such as Slackwater Darter, Etheostoma boschungi, Tuscumbia Darter, E. tuscumbia, and Flame Chub, Hemitremia flammea, over the past several decades, the last comprehensive fish survey of the watershed was conducted in 1971, and changes in the fish fauna and community composition may have occurred. Historical data (1845‐ 2019) containing 5,105 records was compiled for 109 species reported at least once from the Cypress Creek system. Samples from 2009‐19 (primarily in 2015‐16) at 83 localities were collected by seining, backpack electrofishing, and a limited number of boat electrofishing efforts. Community composition was analyzed for 39 collections using the Index of Biotic Integrity. One hundred thirty‐one collections yielded 2,453 records containing a total of 80 species, with a mean of 21 species per collection, including several species of interest. Multiple species (e.g., Pirate Perch, Aphredoderus sayanus) that were rare in historical collections were proportionally documented more frequently in this study when sampling appropriate habitats in select tributaries. The current fish fauna of Cypress Creek includes 99 species, which represents high species diversity for this relatively moderate‐sized stream, when compared to rivers and streams frequently referenced for their fish diversity. Index of Biotic Integrity scores varied among tributaries, suggesting differing anthropogenic impacts on fish community composition across the watershed. Future development within the watershed should be closely monitored to ensure the aquatic biodiversity of this stream is not compromised.