Validation of a nutria (Myocastor coypus) environmental DNA assay highlights considerations for sampling methodology

Abstract Nutria (Myocastor coypus) is a semiaquatic rodent species that is invasive across multiple regions within the United States. Here, we evaluated a qPCR assay previously described for use in Japan for application across invasive populations in the United States. We also compared two environmental DNA sampling methodologies for this assay: field filtration of large volumes of water passed through filters versus direct sampling of small volumes of water. We validated assay specificity, generality, and sensitivity, compared assay performance between two independent laboratories, and successfully tested the assay in situ on a known wild population. The filtration method required fewer samples for environmental DNA detection than direct sampling, but the choice of methods should be assessed based on specific field conditions and time and budget considerations. Our extensive assay validation and comparison across laboratories suggest that the assay is ready to be applied in environmental DNA monitoring of nutria throughout the United States.