Developing Environmental DNA Metabarcoding for the Detection of Elusive Vernal Pool-Breeding Amphibians

Overview

According to the IUCN Red List, 41% of amphibian species worldwide are threatened with extinction. Conservation efforts for amphibian populations are limited as traditional biomonitoring methods fail to accurately census elusive species. This study developed a passive biomonitoring technique, using environmental DNA (eDNA) metabarcoding to target two elusive pool-breeding salamander species, the Jefferson salamander (Ambystoma jeffersonianum) and the blue-spotted salamander (Ambystoma laterale), as well as the ubiquitous wood frog species (Rana sylvatica). Aquatic eDNA was extracted from 102 periodic water grabs from four vernal pools and four brooks/streams in the Hudson River watershed between February to May in 2021 and 2022. The vertebrate 12S rRNA mitochondrial gene was amplified, sequenced, and matched to our novel DNA reference sequences for spotted salamander (Ambystoma maculatum) and Jefferson salamander (Ambystoma jeffersonianum) and GenBank nucleotide data. eDNA metabarcoding detected 63 vertebrate species, including ten amphibian species, along with multiple elusive amphibians: Ambystoma jeffersonianum-laterale complex, spring peeper (Pseudacris crucifer), two-lined salamander (Eurycea bisleanata), and spotted salamander (Ambystoma maculatum). Between the two wetland types surveyed, vernal pool obligate detections were highly specific to vernal pools (95.6%; p < .001), reinforcing eDNA as a proxy for observed species’ presence. The lack of vernal pool protections underscores the urgency for monitoring methods that accurately census elusive species, and the results of this study demonstrate the viability of eDNA metabarcoding as a novel amphibian monitoring technique and its broad applications for the conservation of threatened species.

Competition history

• JSHS 2023 Category not listed

Resources

• National JSHS abstract book ↗

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