Improved Non-Invasive Detection of Cryptobranchus Allegeniensis using Multiplexed Total Nucleic Acid eDNA/eRNA Comparisons

The Eastern Hellbender (Cryptobranchus alleganiensis) is a scarce and reclusive aquatic salamander that resides in swiftly flowing streams within isolated regions of Appalachia. Hellbender populations have been decreasing for decades and Hurricane Helene's recent devastation of Western North Carolina further disrupted population strongholds with reports of the animals displaced and killed due to flash floods in the Hurricane's wake. The recent U.S. Fish and Wildlife request for federal protection under the Endangered Species Act has emphasized the importance of ongoing conservation efforts for the animal, including monitoring and preserving the existing population. Traditional field surveys (TFS) make up the bulk of Hellbender detection efforts although multiple, non-invasive environmental DNA (eDNA) assays have been developed in recent years. Adoption of any eDNA method requires meticulous testing and verification, as some published primer sets for Hellbender have been shown to produce off-target amplification. The goal of this research is to increase the sensitivity and accuracy of eDNA detection methods for the Eastern Hellbender, and to provide an additional molecular target for Ohio stream conservation assessments. This work will consist of developing an RNA molecular marker to examine Eastern Hellbender in conjunction with a DNA marker for Spotted Darter populations through environmental nucleic acids-thus, widening the scope of past Hellbender eDNA assays and providing additional information and confidence in results through eRNA. Both primer sets will be validated by rigorous testing on diverse nucleic acid extractions as performed previously

• Record URL:
  • https://www.dot.state.oh.us/Divisions/Planning/SPR/Research/reportsandplans/Lists/Final%20Reports%20All/Item/displayifs.aspx?List=47f3581d%2Df21c%2D403b%2D9358%2Dfea0b008772b&ID=817&Source=https%3A%2F%2Fwww%2Edot%2Estate%2Eoh%2Eus%2FDivisions%2FPlanning%2FSPR%2FResearch%2Freportsandplans%2FLists%2FFinal%2520Reports%2520All%2FAllItems%2Easpx%3FuseFiltersInViewXml%3D1%26FilterField1%3DCategories%26FilterValue1%3DEnvironmental%26FilterType1%3DMultiChoice%26FilterField2%3DResearch%255Fx0020%255FOrganization%26FilterValue2%3DUniversity%2520of%2520Cincinnati%26FilterType2%3DChoice%26FilterField3%3DProposal%255Fx0020%255FType%26FilterValue3%3DSTAR%26FilterType3%3DChoice&ContentTypeId=0x0100BD006C89430C884FB603A74E63BB6849&RootFolder=%2FDivisions%2FPlanning%2FSPR%2FResearch%2Freportsandplans%2FLists%2FFinal%20Reports%20All

Language

• English

Project

• Status: Active
• Funding: $150,000.00
• Contract Numbers:

  42153

  136989

  123345

• Sponsor Organizations:

  Federal Highway Administration

  1200 New Jersey Avenue, SE
  Washington, DC  United States  20590
• Managing Organizations:

  Ohio Department of Transportation

  Research Program
  1980 West Broad Street
  Columbus, OH  United States  43223
• Project Managers:

  Spriggs, Jennifer

• Performing Organizations:

  University of Cincinnati

  Civil Engineering Department, P.O. Box 210071, 741 Baldwin Hall
  Cincinnati, OH  United States  45221-0071
• Principal Investigators:

  Wendell, David

• Start Date: 20250811
• Expected Completion Date: 20270211
• Actual Completion Date: 0

Subject/Index Terms

• TRT Terms: Amphibians; Detection and identification systems; Endangered species
• Geographic Terms: Ohio
• Subject Areas: Environment; Transportation (General);

Filing Info

• Accession Number: 01957418
• Record Type: Research project
• Source Agency: Ohio Department of Transportation
• Contract Numbers: 42153, 136989, 123345
• Files: RIP, USDOT, STATEDOT
• Created Date: Jun 5 2025 3:01PM