Ambystoma barbouri (Streamside Salamander) inhabits upland deciduous forests associated with ephemeral first- and second-order streams throughout middle Tennessee. The distribution of Streamside Salamanders is centered in north-central Kentucky and extends into adjacent southeastern Indiana and southwestern Ohio. Disjunct populations are known from West Virginia, southern Kentucky, and middle Tennessee. Extant populations for middle Tennessee occur only within the southern Inner Nashville Basin (INB) ecological subregion of the Interior Plateau. Populations were found in Davidson and Jackson counties within the Outer Nashville Basin (ONB) during the 1960s and early 1970s, respectively; however, the status of these populations is unknown. To better determine the distribution of this species in the northern INB and the eastern ONB, we surveyed first- and second-order streams for eggs and larvae. We found Streamside Salamanders at only 2 of 78 localities. One of these breeding sites was adjacent to a previously known breeding stream in Wilson County, and the second site was in Trousdale County in the ONB, approximately 35 km north of the Wilson County sites and 49 km west of the historic Jackson County site. The distribution of Streamside Salamanders is still insufficiently known in middle Tennessee.