A 21st Century butterfly net: Using eDNA to detect the imperiled Dakota skipper

The development of environmental DNA (eDNA) methods for terrestrial arthropods could be transformative for the difficult task of assessing the status of species of conservation concern. The primary goal of this study was to investigate the efficacy of detecting the Dakota skipper (Hesperia dacotae) from its DNA left behind on inflorescences as a means of inferring species presence. We developed and tested a novel qPCR assay and validated the assay in both controlled and field contexts. Using captive animals at the Minnesota Zoo, we found that the number of skippers in an enclosure increased the probability of skipper DNA detection. In the field, Dakota skipper DNA was found on 14% (11 of 81) of inflorescences collected. All detections were from narrowleaf purple coneflower (Echinacea angustifolia). Known visitation of an inflorescence by Dakota skipper prior to sample collection was not a strong predictor of either skipper DNA presence or amount of DNA, but skipper eDNA was detected at 60% (3 of 5) of sites where skippers were observed and 33% (1 of 3) of sites where skippers were not observed. These findings demonstrate successful application of a targeted-species approach to eDNA sampling for butterflies in the field. Taken together, our findings indicate that this method could provide a novel and useful source of data for assessing occupancy trends of butterflies without capturing or even observing them in the wild.