Abstract
The Critically Endangered Smalltooth Sawfish, Pristis pectinata, was once widespread in the tropical and subtropical waters of the Atlantic Ocean, but following substantial declines over the past century, the remaining population(s) are currently confined to Florida in the U.S., and the Bahamas. Recent research and verified public encounter reports suggest that the core population in south and southwest Florida may be stabilizing and potentially expanding into formerly occupied areas of their historic range in the western Atlantic; however, the status of this species outside of core waters is not well understood. Environmental DNA (eDNA) methods provide a relatively cost effective and rapid assessment tool for monitoring species occurrence in aquatic habitats. Here, we have developed an eDNA tool: a species-specific Droplet Digital™ PCR assay targeting a 100-base pair portion of the mitochondrial NADH dehydrogenase subunit 2 gene in P. pectinata, with the ability to reliably detect as little as 0.25 pg of target DNA. The assay was validated by analyzing a water sample from an occupied nursery in southwest Florida, which was found to contain an average of 11.54 copies of target DNA/µL (SE = 0.72) in the reaction. The assay was then further tested by placing a juvenile sawfish in an ex situ tank and analyzing water samples collected at time intervals. The implementation of this eDNA tool into field surveys will provide additional, reliable data to assess species recovery and aid in prioritizing localities in which to focus new research, conservation, and education initiatives.
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10 November 2021
A Correction to this paper has been published: https://doi.org/10.1007/s12686-021-01244-1
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Acknowledgements
This research was funded by The University of Southern Mississippi and was supported by the Mississippi Institutional Development Award (IDeA) Network of Biomedical Research Excellence (INBRE), funded under Institutional Development Award (IDeA) number P20-GM103476 from the National Institutes of General Medical Sciences of the National Institutes of Health. Thank you to Jonathan Lindner for general advice regarding Droplet Digital™ PCR. Thanks to Joshua Speed, London Williams, and Michael Garrett for laboratory access at University of Mississippi Medical Center (UMMC), and for use of Droplet Digital™ PCR equipment. Thanks to Alia Court and Andrew Wooley for field and laboratory support in Florida and to Kevin Feldheim and Kelcee Smith for extracting and providing genomic DNA. Positive sample collection in Florida was supported by funding from the U.S. Department of Commerce, National Oceanic and Atmospheric Administration’s (NOAA) National Marine Fisheries Service through Section 6 (Cooperation with the States) of the U.S. Endangered Species Act under grant award NA16NMF4720062 to the Florida Fish and Wildlife Conservation Commission. Florida sampling was also supported by Keystone Grant 384 to GRP from the Save Our Seas Foundation. Statements, findings, conclusions, and recommendations are those of the authors and do not necessarily reflect the views or policies of the funders. This research was conducted under Endangered Species Permit number 21043 (GRP) issued by NOAA Fisheries.
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Lehman, R.N., Poulakis, G.R., Scharer, R.M. et al. An environmental DNA tool for monitoring the status of the Critically Endangered Smalltooth Sawfish, Pristis pectinata, in the western Atlantic. Conservation Genet Resour 12, 621–629 (2020). https://doi.org/10.1007/s12686-020-01149-5
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DOI: https://doi.org/10.1007/s12686-020-01149-5