Simpsonaias ambigua (Salamander Mussel) is the only North American freshwater mussel known to parasitize an amphibian for reproduction. Prior to this study, the only records of this species from the Cumberland River drainage were from the Stones and Caney Fork River systems, and all records from the Cumberland River drainage are from 1965 or earlier. In 2017, while conducting a survey for freshwater mussels in the Harpeth River, a direct tributary to the Cumberland River, we found 2 fresh-dead Salamander Mussels in Cheatham County, TN, representing the first record of this species from the Harpeth River drainage. Future surveys targeting the specific habit for the Salamander Mussel are recommended to accurately assess its status in the Harpeth River and elsewhere.