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Noninvasive genetic analyses for estimating population size and genetic diversity of the remaining Far Eastern leopard (Panthera pardus orientalis) population

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Abstract

Understanding and monitoring the population status of endangered species is vital for developing appropriate management interventions. We used noninvasive genetic analyses to obtain ecological and genetic data on the last remaining Far Eastern leopard population in the world. During seven winters from 2000–2001 to 2007–2008, we collected feces, hair, and saliva from most of the leopard habitat. Of the 239 leopard samples collected during the study period, 155 were successfully genotyped at 13 microsatellite loci and 37 individuals (18 males and 19 females) were identified. Population size estimates based on the Capwire model were 28 (95 % CI 19–38) in 2002–03 and 26 (95 % CI 13–33) in 2007–2008. The leopard population had a low level of genetic diversity (expected and observed heterozygosity = 0.43; average number of alleles per locus = 2.62), and effective population size was estimated to be low (N e = 7–16) by two genetic-based methods. We observed little improvement in the genetic diversity during the study period and did find an indication of allele loss compared with individuals from the mid-1990s, suggesting that the remaining population will continue to suffer loss of genetic diversity. Given the small population size and the low genetic diversity, with little expectation of replenishment of the genetic variation by natural immigration, successful expansion of available habitat and development of a second population based on captive individuals may be crucial for persistence of this leopard subspecies in the wild.

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References

  • Abramov VK, Pikunov DG (1974) Leopards in the far east of the USSR and their protection. Bull Mosc Soc Nat Amateur Sec Biol 79:5–15 in Russian

    Google Scholar 

  • Adams JR, Vucetich LM, Hedrick PW, Peterson RO, Vucetich JA (2011) Genomic sweep and potential genetic rescue during limiting environmental conditions in an isolated wolf population. Proc R Soc B-Biol Sci 278:3336–3344

    Article  Google Scholar 

  • Aramilev VV, Fomenko PV (2000) Distribution and number of Far Eastern Leopards in the Southwest Primorski Krai. In: Vashukevich YE (ed) Conservation and rational use of animals and floral resources. Irkutsk Agriculture Academy Publishing House, Irkutsk, pp 50–63

    Google Scholar 

  • Ardren WR, Kapuscinski AR (2003) Demographic and genetic estimates of effective population size (N e ) reveals genetic compensation in steelhead trout. Mol Ecol 12:35–49

    Article  CAS  PubMed  Google Scholar 

  • Augustine J, Miquelle DG, Korkishko VG (1996) Preliminary results of the Far Eastern leopard ecology project: implications for conservation and management. Zov Taigi 4:6–11

    Google Scholar 

  • Boom R, Sol CJA, Salimans MMM, Jansen CL, Wertheimvandillen PME, Vandernoordaa J (1990) Rapid and simple method for purification of nucleic acids. J Clin Microbiol 28:495–503

    CAS  PubMed Central  PubMed  Google Scholar 

  • Broquet T, Petit E (2004) Quantifying genotyping errors in noninvasive population genetics. Mol Ecol 13:3601–3608

    Article  CAS  PubMed  Google Scholar 

  • Crow JF, Kimura M (1970) An introduction to population genetics theory. Harper and Row, New York

    Google Scholar 

  • De Barba M, Waits LP, Garton EO, Genovesi P, Randi E, Mustoni A, Groff C (2010) The power of genetic monitoring for studying demography, ecology and genetics of a reintroduced brown bear population. Mol Ecol 19:3938–3951

    Article  PubMed  Google Scholar 

  • Frankham R (1995) Effective population size/adult population size ratios in wildlife: a review. Genet Res 66:95–107

    Article  Google Scholar 

  • Goudet J (2001) FSTAT, a program to estimate and test gene diversities and fixation indices

  • Hebblewhite M, Miguelle DG, Murzin AA, Aramilev VV, Pikunov DG (2011) Predicting potential habitat and population size for reintroduction of the Far Eastern leopards in the Russian Far East. Biol Conserv 144:2403–2413

    Article  Google Scholar 

  • Höss M, Pääbo S (1993) DNA extraction from Pleistocene bones by a silica-based purification method. Nucl Acids Res 21:3913–3914

    Article  PubMed Central  PubMed  Google Scholar 

  • Jenny D (1996) Spatial organization of leopards Panthera pardus in Tai National Park, Ivory Coast: is rain forest habitat a ‘tropical haven’? J Zool 240:427–440

    Article  Google Scholar 

  • Keller LF, Waller DM (2002) Inbreeding effects in wild populations. Trends Ecol Evol 17:230–241

    Article  Google Scholar 

  • Kerley LL (2010) Using dogs for tiger conservation and research. Integr Zool 5:390–396

    Article  PubMed  Google Scholar 

  • Kimura M, Crow JF (1964) The number of alleles that can be maintained in a finite population. Genetics 49:725–738

    CAS  PubMed Central  PubMed  Google Scholar 

  • Koelewijn HP, Perez-Haro M, Jansman HAH, Boerwinkel MC, Bovenschen J, Lammertsma DR, Niewold FJJ, Kuiters AT (2010) The reintroduction of the Eurasian otter (Lutra lutra) into the Netherlands: hidden life revealed by noninvasive genetic monitoring. Conserv Genet 11:601–614

    Article  Google Scholar 

  • Kostyria AV, Skorodelov AS, Miquelle DG, Aramilev VV, McCullough DR (2003) Results of camera trap survey of Far Eastern leopard population in southwest Primorski Krai, winter 2002–2003. Wildlife Conservation Society, Institute of Sustainable Use of Nature Resources, University of California (Berkeley), Vladivostok, Russia

  • Lewis J (2010) Disease management program. In: A program for reintroduction of the Far Eastern leopard into southern Sikhote-Alin, Primorskii Krai, Russian Far East. http://www.wcsrussia.org/en-us/publications/thefareasternleopardscientificpublications.aspx. Accessed 1 March 2010

  • MacKay P, Smith DA, Long RA, Parker M (2008) Scat detection dogs. In: Long RA, MacKay P, Zielinski WJ, Ray JC (eds) Noninvasive survey methods for carnivores. Island Press, Washington, pp 183–222

    Google Scholar 

  • Marker LL, Dickman AJ (2005) Factors affecting leopard (Panthera pardus) spatial ecology, with particular reference to Namibian farmlands. S Afr J Wildl Res 35:105–115

    Google Scholar 

  • McKelvey KS, Schwartz MK (2004) Genetic errors associated with population estimation using non-invasive molecular tagging: problems and new solutions. J Wildl Manage 68:439–448

    Article  Google Scholar 

  • Menotti-Raymond M, David VA, Lyons LA, Schaffer AA, Tomlin JF, Hutton MK, O’Brien SJ (1999) A genetic linkage map of microsatellites in the domestic cat (Felis catus). Genomics 57:9–23

    Article  CAS  PubMed  Google Scholar 

  • Miller CR, Joyce P, Waits LP (2005) A new method for estimating the size of small populations from genetic mark-recapture data. Mol Ecol 14:1991–2005

    Article  CAS  PubMed  Google Scholar 

  • Miquelle DG, Murzin AA (2000) Spatial distribution of Far Eastern leopard in Southwest Primorski Krai, and recommendations for their conservation. Wildlife Conservation Society, World Wildlife Fund, and Tigris Foundation, Vladivostok

  • Miquelle DG, Pikunov DG (2003) Status of the Amur tiger and Far Eastern leopard in Northeast Asia. In: Newell JP (ed) The Russian Far East: a reference guide for conservation and development. Daniel & Daniel Publishers Inc., McKinleyville, pp 106–109

    Google Scholar 

  • Miquelle DG, Pikunov DG, Dunishenko YM, Aramilev VV, Nikolaev IG, Abramov VK, Smirnov EN, Salkina GP, Matyushkin EN, Murzin AA (2006) A theoretical basis for surveys of Amur tigers and their prey base in the Russian Far East. DalNauka, Vladivostok in Russian

    Google Scholar 

  • Mizutani F, Jewell PA (1998) Home-range and movements of leopards (Panthera pardus) on a livestock ranch in Kenya. J Zool 244:269–286

    Article  Google Scholar 

  • Nagata J, Aramilev VV, Belozor A, Sugimoto T, McCullough DR (2005) Fecal genetic analysis using PCR-RFLP of cytochrome b to identify sympatric carnivores, the tiger Panthera tigris and the leopard Panthera pardus, in Far Eastern Russia. Conserv Genet 6:863–866

    Article  Google Scholar 

  • Nowell K, Jackson P (1996) Wild cats: status survey and conservation management plan. IUCN/SSC Cat Specialist Group, Gland

    Google Scholar 

  • Odden M, Wegge P (2005) Spacing and activity patterns of leopards Panthera pardus in the Royal Bardia National Park, Nepal. Wildl Biol 11:145–152

    Article  Google Scholar 

  • Paetkau D (2003) An empirical exploration of data quality in DNA-based population inventories. Mol Ecol 12:1375–1387

    Article  CAS  PubMed  Google Scholar 

  • Pikunov DG, Korkishko VG (1985) Modern distribution and numbers of the leopard (Panthera pardus) in the Soviet Far East. Zool J 64:897–905 (in Russian)

    Google Scholar 

  • Pikunov DG, Miquelle DG, Abramov VK, Nikolaev IG, Seredkin IV, Murzin AA, Korkishko VG (2003) A survey of Far Eastern leopard and Amur tiger populations in Southwest Primorski Krai, Russian Far East (February 2003). Pacific Institute of Geography (FEB RAS), Wildlife Conservation Society, and Tigris Foundation, Vladivostok

  • Pikunov DG, Seredkin IV, Aramilev VV, Nikolaev IG, Murzin AA (2009) Numbers of Far Eastern leopards (Panthera pardus orientalis) and Amur Tigers (Panthera tigris altaica) in Southwest Primorski Krai, Russian Far East, 2007. DalNauka, Vladivostok

    Google Scholar 

  • Pocock RI (1930) The panthers and ounces of Asia. J Bombay Nat Hist Soc 34:307–336

    Google Scholar 

  • Pritchard JK, Stephens M, Donnelly P (2000) Inference of population structure using multilocus genotype data. Genetics 155:945–959

    CAS  PubMed Central  PubMed  Google Scholar 

  • Rabinowitz A (1989) The density and behaviour of large cats in a dry tropical forest mosaic in Huai Kha Khaeng wildlife sanctuary, Thailand. Nat His Bull Siam Soc 37:235–251

    Google Scholar 

  • Raymond M, Rousset F (1995) Genepop (Version-1.2)—population-genetics software for exact tests and ecumenicism. J Hered 86:248–249

    Google Scholar 

  • Rice WR (1989) Analyzing tables of statistical tests. Evolution 43:223–225

    Article  Google Scholar 

  • Salmanova E (2008) Territorial use of Far Eastern leopards. M.S. thesis, Far Eastern State University, Primorskii Krai, Russia

  • Salmanova E (2011) Characteristics of scrapes of Far Eastern leopards as a primary means of scentmarking. In: Materials from the X conference of students from universities and research organizations of the Russian Far East: problems in ecology, marine biology, and biotechnology, pp 244–245

  • Schwartz MK, Luikart G, Waples RS (2007) Genetic monitoring as a promising tool for conservation and management. Trends Ecol Evol 22:25–33

    Article  PubMed  Google Scholar 

  • Stander PE, Haden PJ, Kaqece, Ghau (1997) The ecology of a sociality in Namibian leopards. J Zool 242:343–364

    Article  Google Scholar 

  • Sugimoto T, Nagata J, Aramilev VV, Belozor A, Higashi S, McCullough DR (2006) Species and sex identification from faecal samples of sympatric carnivores, Amur leopard and Siberian tiger, in the Russian Far East. Conserv Genet 7:799–802

    Article  Google Scholar 

  • Sugimoto T, Nagata J, Aramilev VV, McCullough DR (2012) Population size estimation of Amur tigers in Russian Far East using noninvasive genetic samples. J Mamm 93:93–101

    Article  Google Scholar 

  • Sun B, Miquelle DG, Xiaochen Y, Zhang E, Hiyai S, Goshen G, Pikunov DG, Dunishenko YM, Nikolaev IG, Daming L (1999) Survey of Amur tigers and Far Eastern leopards in eastern Heilongjiang Province, China, and recommendations for their conservation. Final report to the Wildlife Conservation Society, Vladivostok, Russia

  • Sunquist M, Sunquist F (2002) Wild cats of the world. The University of Chicago Press, Chicago

    Google Scholar 

  • Taberlet P, Waits LP, Luikart G (1999) Noninvasive genetic sampling: look before you leap. Trends Ecol Evol 14:323–327

    Article  PubMed  Google Scholar 

  • Tallmon DA, Koyuk A, Luikart G, Beaumont MA (2008) ONeSAMP: a program to estimate effective population size using approximate Bayesian computation. Mol Ecol Resour 8:299–301

    Article  PubMed  Google Scholar 

  • Uphyrkina O, Miquelle D, Quigley H, Driscoll C, O’Brien SJ (2002) Conservation genetics of the Far Eastern leopard (Panthera pardus orientalis). J Hered 93:303–311

    Article  CAS  PubMed  Google Scholar 

  • Valière N (2002) GIMLET: a computer program for analysing genetic individual identification data. Mol Ecol Notes 2:377–379

    Article  Google Scholar 

  • Van Oosterhout C, Hutchinson WF, Wills DPM, Shipley P (2004) MICRO-CHECKER: software for identifying and correcting genotyping errors in microsatellite data. Mol Ecol Notes 4:535–538

    Article  Google Scholar 

  • Waits LP, Paetkau D (2005) Noninvasive genetic sampling tools for wildlife biologists: a review of applications and recommendations for accurate data collection. J Wildl Manage 69:1419–1433

    Article  Google Scholar 

  • Waits LP, Luikart G, Taberlet P (2001) Estimating the probability of identity among genotypes in natural populations: cautions and guidelines. Mol Ecol 10:249–256

    Article  CAS  PubMed  Google Scholar 

  • Waples RS, Do C (2008) LDNE: a program for estimating effective population size from data on linkage disequilibrium. Mol Ecol Resour 8:753–756

    Article  PubMed  Google Scholar 

  • Wasser SK, Houston CS, Koehler GM, Cadd GG, Fain SR (1997) Techniques for application of faecal DNA methods to field studies of Ursids. Mol Ecol 6:1091–1097

    Article  CAS  PubMed  Google Scholar 

  • Wildlife Conservation Society (2012) Rare appearance for an extremely rare Cat. http://www.wcs.org/news-and-features-main/rare-appearance-for-extremely-rare-cat.aspx. Accessed 25 April 2012

  • Yang S, Jiang J, Wu Z, Li T, Yang X, Han X, Miquelle DG, Pikunov DG, Dunishenko YM, Nikolaev IG (1998) A survey of tigers and leopards in eastern Jilin Province, China, winter 1998. Final report to the UNDP and the Wildlife Conservation Society, Vladivostok, Russia

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Acknowledgments

We thank the Kobe Oji Zoo, Hiroshima City Asa Zoological Park, Asahiyama Zoo, and S. J. O’Brien (the National Cancer Institute, USA) for the Far Eastern leopard DNA samples, and Inokashira Zoo, R. Masuda, and Ueno Zoological Gardens for the leopard cat DNA samples. We also thank M. Stuewe, Y. Uryu, K. Kobyakov, Y. Darman, O. Walter, and S. Christie for providing us with valuable information on the Far Eastern leopard; and the WWF Russian Far East office for providing us with maps and information on the forest restoration project. We express special gratitude to A. Belozor and S. Higashi for their assistance in the realization of this project. Financial support for this study was provided by the A. Starker Leopold Endowed Chair, University of California, Berkeley (D. McCullough, chair-holder) and a Grant-in-Aid for JSPS Fellows (19-05514) from Japan Society for the Promotion of Science and in part by WWF US.

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Authors and Affiliations

  1. Faculty of Environmental Earth Science, Hokkaido University, Sapporo, Hokkaido, 0600810, Japan

    Taro Sugimoto

  2. Pacific Institute of Geography, Far Eastern Branch of the Russian Academy of Sciences, Vladivostok, 690041, Russia

    Vladimir V. Aramilev

  3. Zoological Society of London, Regent’s Park, London, UK

    Linda L. Kerley

  4. Forestry and Forest Products Research Institute, Tsukuba, 3058687, Japan

    Junco Nagata

  5. Russia Program, Wildlife Conservation Society, Bronx, NY, 10460, USA

    Dale G. Miquelle

  6. Department of Environmental Science, Policy, and Management, and Museum of Vertebrate Zoology, University of California, Berkeley, Berkeley, CA, 94720, USA

    Dale R. McCullough

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  1. Taro Sugimoto
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  2. Vladimir V. Aramilev
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  5. Dale G. Miquelle
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Correspondence to Taro Sugimoto.

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Sugimoto, T., Aramilev, V.V., Kerley, L.L. et al. Noninvasive genetic analyses for estimating population size and genetic diversity of the remaining Far Eastern leopard (Panthera pardus orientalis) population. Conserv Genet 15, 521–532 (2014). https://doi.org/10.1007/s10592-013-0558-8

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