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January 17, 2020 Miguel GarcÃa-Bermúdez Fish and Wildlife Administrator Science Application, U.S. Fish & Wildlife Service South Atlantic, Gulf and Mississippi-Basin San Juan, Puerto Rico Dear Dr. GarcÃa-Bermúdez, This letter describes the work and transmits the final evaluation of applications to carry out research to determine the taxonomy of wild canid populations in regions of the United States where recent evidence suggests the potential presence of red wolves. BACKGROUND In 1980, the red wolf (Canis rufus) was declared extinct in the wild. However, there have been several recent reports of âred wolfâ sightings in southwestern Louisiana and Texas. Preliminary evidence suggests that these wild canines, originally thought to be coyotes, may at a minimum carry some red wolf alleles or, at a maximum, may be overlooked populations of wild red wolves. Genetic analyses of two deceased animals on Galveston Island, Texas, revealed shared alleles with the captive breeding red wolf population in North Carolina and also some genetic variation that is distinct from any known wild North American canine.1 Analyses of DNA microsatellite variation at 9 nuclear loci and mitochondrial DNA (mtDNA) from wild canine scat, hair, and tissue samples from southwestern Louisiana suggest that several animals have more than 40 percent red wolf ancestry, and at least one animal has 100 percent red wolf ancestry at the 9 microsatellite loci and mtDNA.2 Because the red wolf is a critically endangered species, determining the identity of the wild canines in southern Louisiana and adjacent areas is a top priority for the U.S. Fish & Wildlife Service (FWS). FWS commissioned the National Academies of Sciences, Engineering, and Medicine (the National Academies) to develop a request for applications (RFA), âResearch to determine the taxonomy of wild canid populations in regions of the United States where recent evidence suggests the potential presence of red wolves (Canis rufus),â and to conduct an independent evaluation of the submitted applications. The RFA defined two project goals: (1) to collect and analyze morphological, genetic, and genomic data to clarify the taxonomic identity of the wild canid populations in southwestern Louisiana and other geographic areas in the southeastern United States that red wolves were historically known to inhabit, and (2) if feasible, to deliver results to FWS within 8 to 12 months from the receipt of funding. Applicants were asked to describe study designs for the collection of genetic, genomic, and morphological data; describe analyses; assess the availability of appropriate outgroup genomes; propose a clear plan for 1 Heppenheimer, E., K. E. Brzeski, R. Wooten, W. Waddell, L. Y. Rutledge, M. J. Chamberlain, D. R. Stahler, J. W. Hinton, and B. M. vonHoldt. 2018. Rediscovery of red wolf ghost alleles in a canid population along the American Gulf Coast. Genes (Basel) 9(1):e618. 2 Murphy, S. M., J. R. Adams, J. J. Cox, and L. P. Waits. 2019. Substantial red wolf genetic ancestry persists in wild canids of southwestern Louisiana. Conservation Letters 12(2):e12621. 3
sampling animals without harm; and provide a statement of intent for publishing results and making data available to the public. To carry out this work, the National Academies appointed a nine-person committee, chaired by Dr. Joseph Travis of the Florida State University. The committee membersâ collective expertise includes conservation biology, genomic and other kinds of genetic analyses, morphology and paleontology, and the genetics and evolution of canids. The committee evaluated the scientific merit of the proposed research and provided additional scientific and technical factors for FWS to consider. The selection of applications for funding will be determined by FWS. EVALUATION PROCESS Each committee member was responsible for writing a brief summary assessment of how well each application met the criteria of the RFA before holding a discussion in a committee meeting. Prior to their evaluation of the applications, all of the committee members were screened for potential financial conflicts of interests and relevant relationships with organizations and individuals involved with the submitted applications. None of the committee members responded to the RFA. In cases in which a committee member had a financial or research association with an applicant, the committee member did not review that individualâs application. Five criteria were used to evaluate each application: 1. Efficiency and thoroughness of the sampling plan. 2. Appropriateness of the proposed genomic and morphological methods. 3. Power of the proposed analyses. 4. Potential to provide results to FWS within 8 to 12 months from the receipt of the award. 5. Qualifications and experience of principal investigators and senior personnel. The committee held its evaluation meeting in Washington, DC, on December 8â9, 2019. At the meeting, one member first outlined the strengths and weaknesses of each application. Next, the committee held a plenary discussion of the scientific merit (criteria 1â3), timeline, and the qualifications of the principal investigator and key personnel in order to come to a consensus on which applications best met the selection criteria and would best assess the identity of the canid populations in southwestern Louisiana and southeastern Texas. The committeeâs summary evaluation of the three applications and cross-cutting considerations are appended to this letter report as non-public Appendixes B and C. CONCLUSIONS AND RECOMMENDATION The committee received three applications in response to the RFA. No single application would unambiguously resolve the taxonomic status of the canid populations in the southeastern United States. However, aspects of two of the applications were of high scientific merit and could together yield reliable conclusions about red wolf identity in the canids in southwestern Louisiana and southeastern Texas. One of these applications was comprehensive and addressed each element of the RFA. However, the committee is concerned that the proposed genomic 4
methods in this application may yield uncertain results and that the proposed approach to resolve some of the uncertainties will significantly exceed the timeframe within which FWS expects to receive results. A second application, though scientifically comprehensive, focused only on defining genomic markers associated with the red wolf. In the second application, the principal investigators proposed to create more robust genomic reference data than currently exists, which will be critical for the unambiguous interpretation of genomic data on the unidentified canids in the southeastern United States. The work in the second application could be completed well in advance of any genomic analysis of field-collected samples. Therefore, the committee judged that the narrower focus of the application was not a reason to disqualify it from consideration. The committee recommends that FWS fund specific aspects of each of these two applications that, in combination, present a strategic and robust scientific approach to addressing taxonomic questions about the unidentified canids in southwestern Louisiana and southeastern Texas. The collective results of other aspects will also accelerate further research on canid ancestry. Though some proposed activities in these two applications are not essential to achieve the goals described in the RFA, FWS will be more likely to obtain answers to its questions if it provides funding for the specific aspects relevant to the RFA in both applications as described in the non-public appendixes. The committee would like to thank all of the applicants for responding to the RFA at such short notice. It is anticipated that FWS will request full research proposals from the applicants, and it will allocate the funds to complete the work, taking into consideration the committeeâs evaluation and internal agency factors. We hope that FWS will find the committeeâs recommendations and comments useful in its efforts and look forward to the outcome of the full proposal process. Sincerely, Joseph Travis Chair, Committee on Assistance to the U.S. Fish & Wildlife Service on Taxonomic Studies of the Red Wolf: A Review of Applications to Carry Out Research and Development of a Research Strategy cc: Mallory Martin, Coordinator for the Southeast Conservation Adaptation Strategy and the South Atlantic Blueprint, U.S. Fish & Wildlife Service 5
APPENDIXES A. List of Received Applications* B. Evaluations of the Individual Applications* C. Cross-Cutting Considerations* D. Request for Applications E. Committee Roster F. Acknowledgment of Report Reviewers *This appendix is not available to members of the public. 6
