Development of microsatellite markers and a restriction endonuclease digest assay for non-invasive sampling of endangere
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TECHNICAL NOTE
Development of microsatellite markers and a restriction endonuclease digest assay for non-invasive sampling of endangered White-rumped, Slender-billed and Red-headed vultures Y. A. Kapetanakos • I. J. Lovette • T. E. Katzner
Received: 5 March 2014 / Accepted: 27 March 2014 Ó Springer Science+Business Media Dordrecht 2014
Abstract Southeast Asian vultures have been greatly reduced in range and population numbers, but it is challenging to use traditional tagging and monitoring techniques to track changes in their populations. Genotypes derived from non-invasively collected feather samples provide an alternative and effective means to ‘capture’ individual vultures for mark-recapture analyses. We describe a restriction endonuclease digest assay that distinguishes the visually similar feathers of three species of critically endangered Asian vultures (Gyps bengalensis, G. tenuirostris, and Sarcogyps calvus). In addition, we describe a panel of eight polymorphic microsatellite loci. In combination, the restriction endonuclease assay and microsatellite marker set developed here are powerful molecular tools for investigating the genetic and demographic status of these Asian vultures species.
Y. A. Kapetanakos I. J. Lovette Department of Ecology and Evolutionary Biology, Cornell University, Ithaca, NY, USA e-mail: [email protected] Y. A. Kapetanakos (&) I. J. Lovette Fuller Evolutionary Biology Program, Cornell Lab of Ornithology, 159 Sapsucker Woods Rd., Ithaca, NY 14850, USA e-mail: [email protected] T. E. Katzner Division of Forestry and Natural Resources, West Virginia University, Percival Hall, Room 307D, PO Box 6125, Morgantown, WV 26506, USA e-mail: [email protected] T. E. Katzner USDA Forest Service, Northern Research Station, Parsons, WV 26287, USA
Keywords Gyps bengalensis Gyps tenuirostris Sarcogyps calvus Microsatellite Cambodia Of the nine species of vulture found in Asia, seven are undergoing population declines (IUCN 2013) largely due to severe reduction in food resources, habitat loss, and poisoning (Clements et al. 2013; Pain et al. 2003). Within the last two decades, three species found on the Indian subcontinent, White-rumped (Gyps bengalensis), Slender-billed (G. tenuirostris) and Red-headed (Sarcogyps calvus) vultures, were nearly extirpated by secondary exposure to the veterinary pharmaceutical diclofenac (Green et al. 2004) and are now listed as critically endangered (IUCN 2013). The three species also occur in low population densities in Cambodia. Monitoring and research efforts to conserve the Cambodian populations are underway through visual surveys and supplemental feeding programs that began in 2004 (Clements et al. 2013). To estimate abundance and genetic variability, we initiated a non-invasive genetic mark-recapture study using naturally shed feathers in 2008 (e.g. Rudnick et al. 2008). We developed an endonuclease digest assay to identify the vulture species associated with each feather sample and generated a panel of microsatellite markers to identify uni
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