Application of environmental DNA detection for surveillance of Four-toed salamander

We evaluated the efficacy of eDNA monitoring for the four-toed salamander (Hemidactylium scutatum), a small terrestrial salamander that is threatened throughout much of its range. We collected aquatic eDNA weekly for 8 weeks from vernal pools with historical reports of H. scutatum presence (n = 6) and contemporary reports of H. scutatum adults on associated pool uplands (n = 4). We confirmed specificity of qPCR for eDNA by sequencing qPCR amplicons.