Global population and species-level amphibian declines are attributable to multiple environmental and biological factors including the disease chytridiomycosis caused by the chytrid fungus, Batrachochytrium dendrobatidis (Bd). In North America, chytridiomycosis-mediated declines may be severe, but the occurrence of Bd is also patchy. The Southern Appalachian Mountains are a global hotspot for salamander diversity, yet relatively few surveys have focused on the prevalence of Bd in salamanders. From 2008 to 2013, we collected 668 swabs from 603 individual amphibians (some were captured and swabbed twice) of 43 species (seven Anura and 36 Caudata) from the Southeastern Piedmont and Southern Appalachian Mountains in western North Carolina and northeastern Tennessee. We used replicate PCR-assays and found that Bd was present but extremely uncommon (1.00%) in salamanders of the region and was not detected at all in the four anuran taxa sampled. We detected six Bd-positive salamanders, including five Spotted Salamanders (Ambystoma maculatum; 10% of individuals sampled) from Watauga County, North Carolina, and one Green Salamander (Aneides aeneus; 7.7% of individuals sampled) from Transylvania County, North Carolina. Collectively, our data suggest that Bd is very uncommon in this salamander hotspot. Thus, Bd may not be a cause of current and future declines in this region. These data serve as an important baseline for future studies of amphibian abundance, distribution, and assemblage structure in this region.