Larvae of Neophylax toshioi Vineyard and Wiggins were first collected from Doe Creek, Johnson County, Tennessee, in 2005 during a qualitative benthic survey conducted by the Tennessee Wildlife Resources Agency. Additional collections in 2007 allowed confirmation of the species by association of adult with larval stages through a series of eight reared male and female metamorphotype pupae. Larvae were again collected from the Doe Creek site and in nearby Roan Creek in early 2007. These findings represent new distribution records for N. toshioi and the first reports of this species from Tennessee.