Diet analysis is essential to fully understanding the biology of a species and its function within the ecosystem, and is also key in identifying food web interactions and the population dynamics of predators and prey. There is currently little information on the diet of small to mid-sized carnivores due to their elusive behavior. We used a DNA-based method to analyze the vertebrate prey of the Pallas’s cat (Otocolobus manul) in the Gongga Mountain Nature Reserve of southwestern China, a global biodiversity hotspot region. We confirmed 14 scat samples containing the Pallas’s cat DNA from 100 small carnivore scats collected at altitudes between 3200 and 4900 m, and then performed polymerase chain reaction (PCR) amplifications using primers targeting the vertebrate mitochondrial 12S rRNA gene with a blocking oligonucleotide specific to the predator sequence. We identified a total of 18 prey taxa, including nine mammals, eight birds and one fish. Our results show that pikas (in 79% of the scats) and birds (in 50% of the scats) are the predominant prey consumed by the Pallas’s cat in our investigation area. The most important finding in our study is that Cypriniformes species were found in the Pallas’s cat’s diet, exhibiting dietary diversification consistent with the accessible fauna of the surrounding environment. In order to increase the detectability of both typical and rare vertebrate prey items, we constructed a blocking primer to inhibit the amplification of the Pallas’s cat DNA.
We are grateful to Xiaohong Sun, Jie Gao, Gai Luo, Xingcheng He and Meiling Cheng for their kind help and suggestions in conducting the research and data analyses. We also thank Professor Yongjie Wu, Mike Seitz and two additional anonymous reviewers for their valuable comments on the manuscript. This study was funded by the Sichuan Gongga Mountain National Nature Reserve central finance forestry protection 2014 subsidy fund snow leopard investigation project.
Competing interests: The authors declare that they have no competing interests.
Availability: The datasets used in the present study are available from the corresponding author on reasonable request.
Consent for publication: Applicable.
Ethics approval and consent to participate: The experiments comply with the current laws of China in which they were performed.
Barashkova, A. and I. Smelansky. 2011. Pallas’s cat in the Altai Republic Russia[J]. Cat News 54: 1–7. Search in Google Scholar
Boyles, J.G. and J.J. Storm. 2007. The perils of picky eating: dietary breadth is related to extinction risk in insectivorous bats. PLoS One 2: e672. Search in Google Scholar
Brownstein, M.J., J.D. Carpten and J.R. Smith. 1996. Modulation of non-templated nucleotide addition by Taq polymerase: primer modification that facilitate genotyping. Bio Techniques 20: 1004–1010. Search in Google Scholar
Carreon-Martinez, L., T.B. Johnson, S.A. Ludsin and D.D. Heath. 2011. Utilization of stomach content DNA to determine diet diversity in piscivorous fishes. J. Fish Biol. 78: 1170–1182. Search in Google Scholar
Deagle, B.E., D.J. Tollit, S.N. Jarman, M.A. Hindell, A.W. Trites and N.J. Gales. 2005. Molecular scatology as a tool to study diet: analysis of prey DNA in scats from captive Steller sea lions. Mol. Ecol. 14: 1831–1842. Search in Google Scholar
Farrell, L.E., J. Roman and M.E. Sunquist. 2000. Dietary separation of sympatric carnivores identified by molecular analysis of scats. Mol. Ecol. 9: 1583–1590. Search in Google Scholar
Ficetola, G.F., E. Coissac and S. Zundel. 2010. An in silico approach for the evaluation of DNA barcodes. BMC Genomics 11: 434. Search in Google Scholar
Fox, J.L. and T. Dorji. 2007. High elevation record for occurrence of the manul or Pallas cat on the northwestern Tibetan plateau, China. Cat News 46: 35. Search in Google Scholar
Harcourt, A.H., S.A. Coppeto and S.A. Parks. 2002. Rarity, specialization and extinction in primates. J. Biogeogr. 29: 44–456. Search in Google Scholar
He, L. and Y. Tang. 2008. Soil development along primary succession sequences on moraines of Hailuogou Glacier, Gongga Mountain, Sichuan, China. Catena 72: 259–269. Search in Google Scholar
Hyunbin J., S.J. Kwang, S.K. Heui and J.J. Gea. 2014. Application of DNA barcoding for identification of freshwater carnivorous fish diets: is number of prey items dependent on size class for Micropterus salmoides? Ecol. Evol. 4: 219–229. Search in Google Scholar
Janecka, J.E., R. Jackson, Y. Zhang, D. Li, B. Munkhtsog, B.V. Buckley and W.J. Murphy. 2008. Population monitoring of snow leopards using noninvasive collection of seat samples: a pilot study. Anim. Conserv. 11: 401–411. Search in Google Scholar
Jarman, S.N., B.E. Deagle and N.J. Gales. 2004. Group-specific polymerase chain reaction for DNA-based analysis of species diversity and identity in dietary samples. Mol. Ecol. 13: 1313–1322. Search in Google Scholar
Jutzeler, E., Y. Xie and K. Vogt. 2010. The smaller felids of China: Pallas’s cat Otocolobus manul. Cat News Special Issue 5: 37–39. Search in Google Scholar
Kageyama, T., Y. Sato, S. Nishizawa and N. Teramae. 2008. Competitive binding of small ligands to nucleobases in AP site-containing DNA duplexes. Nucleic Acids Symp. Ser. 52: 119–120. Search in Google Scholar
King, R.A., D.S. Read, M. Traugott and W. Symondson. 2008. Molecular analysis of predation: a review of best practice for DNA-based approaches. Mol. Ecol. 17: 947–963. Search in Google Scholar
King, R.A., I.P. Vaughan, J.R. Bell, D.A. Bohan and W. Symondson. 2010. Prey choice by carabid beetles feeding on an earthworm community analysed using species and lineage specific PCR primers. Mol. Ecol. 19: 1721–1732. Search in Google Scholar
Li, M.J., Y. Sato, S. Nishizawa, T. Seino, K. Nakamura and N. Teramae. 2009. 2-Aminopurinemodified abasic site containing duplex DNA for highly selective detection of theophylline. J. Am. Chem. Soc. 131: 2448–2449. Search in Google Scholar
Mckinney, M.L. 1997. Extinction vulnerability and selectivity: combining ecological and paleontological views. Ann. Rev. Ecol. Syst. 28: 495–516. Search in Google Scholar
Munkhtsog, B. 2016. “Otocolobus manul”. The IUCN Red List of Threatened Species. IUCN. 2016: e. T15640A87840229. Doi: 10. 2305/ IUCN. UK. 2016-1. RLTS. T15640A87840229. en. Retrived 15 January 2018. Search in Google Scholar
Riaz, T., W. Shehzad, A. Viari, F. Pompanon, P. Taberl and E. Coissac. 2011. ecoPrimers: inference of new DNA barcode markers from whole genome sequence analysis. Nucleic Acids Res. 39: e145. Search in Google Scholar
Ross, S., S. Harris and B. Munkhtsog. 2007. The spatial ecology and conservation of Pallas cat (Otocolobus manul) in central Mongolia. In: (J. Hughes and R. Mercer, eds.) Felid biology and conservation conference 17–20 September: Abstracts, Oxford, UK. pp. 59. Search in Google Scholar
Ross, S., R. Kamnitzer, B. Munktsog and S. Harris. 2010a. Den selection is critical for Pallas’s cats (Otocolobus manul). Can. J. Zool. 88: 905–913. Search in Google Scholar
Ross, S., B. Munktsog and S. Harris. 2010b. Dietary composition, plasticity and prey selection of Pallas’s cats. J. Mammal. 91: 811–817. Search in Google Scholar
Ross, S., B. Munktsog and S. Harris. 2012. Determinants of mesocarnivore range use: relative effects of prey and habitat properties on Pallas’s cat home-range size. J. Mammal. 93: 1292–1300. Search in Google Scholar
Ross, S., A. Barashkova, M.S. Farhadinia, A. Appel, P. Riordan, J. Sanderson and B. Munkhtsog. 2016. “Otocolobus manul”. The IUCN Red List of Threatened Species. IUCN. 2016: e. T15640A87840229. DOI: 10.2305/IUCN.UK.2016-1.RLTS.T15640A87840229.en. Retrieved 15 January 2018. Search in Google Scholar
Shehzad, W., T. Riaz, M.A. Nawaz, C. Miquel, C. Poillot, S.A. Shah, F. Pompanon, E. Coissac and P. Taberlet. 2012. Carnivore diet analysis based on next-generation sequencing: application to the leopard cat (Prionailurus bengalensis) in Pakistan. Mol. Ecol. 21: 1951–1965. Search in Google Scholar
Sheppard, S.K. and J.D. Harwood. 2005. Advances in molecular ecology: tracking trophic links through predator-prey food webs. Funct. Ecol. 19: 751–762. Search in Google Scholar
Smith, A.T. and J.M. Foggin. 1999. The plateau pika (Ochotona curzoniae) is a keystone species for biodiversity on the Tibetan plateau. Anim. Conserv. 2: 235–240. Search in Google Scholar
Smith, A.T., Y. Xie, R. Hoffman, D. Lunde, J. MacKinnon, D.E. Wilson and W.C. Wozencraft. 2008. A guide to the Mammals of China. Princeton University Press, Princeton, New Jersey. Search in Google Scholar
Sunquist, M.E. and F. Sunquist. 2002. Wild cats of the world. University of Chicago Press, Chicago, IL. Search in Google Scholar
Symonds, W. 2002. Molecular identification of prey in predator diets. Mol. Ecol. 11: 627–641. Search in Google Scholar
Titov, A. 2000. Notes on calicioid lichens and fungi from the Gongga Mountains (Sichuan, China). Lichenologist 32: 553–569. Search in Google Scholar
Titov, A. 2001. Further notes on calicioid lichens and fungi from the Gongga Mountains (Sichuan, China). Lichenologist 33: 303–314. Search in Google Scholar
Vestheim, H. and S.N. Jarman. 2008. Blocking primers to enhance PCR amplification of rare sequences in mixed samples-A case study on prey DNA in Antarctic krill stomachs. Front. Zool. 5: 12. Search in Google Scholar
Wang, Y.F., M.T.T. Ng, T.Y. Zhou, X. Li, C.H. Tan, T. Li. 2008. C3-Spacer-containing circular oligonucleotides as inhibitors of human topoisomerase I. Bioorg. Med. Chem. Lett. 18: 3597–3602. Search in Google Scholar
Wang, X.Q., G.H. Wang, F. Qiao, Q.K. Gao, L.H. Kong, Z.R. Zhu and J.A. Chen. 2017. Progress on high-throughput sequencing and its applications in food web analysis. Acta Ecol. Sin. 37: 2530–2539. Search in Google Scholar
Wilson, D.E. and D.M. Reeder. 2005. Mammal species of the world: a taxonomic and geographic reference. 3rd ed. Johns Hopkins University Press, Baltimore, MD, USA. pp. 2142. Search in Google Scholar
Wilson, M.C. and A.T. Smith. 2015. The pika and the watershed: the impact of small mammal poisoning on the ecohydrology of the Qinghai-Tibetan Plateau. AMBIO 44: 16–22. Search in Google Scholar
Xiong, M.Y., X.N. Shao, Y. Long, H.L. Bu, D. Zhang, D.J. Wang, S. Li, R.J. Wang and M. Yao. 2016. Molecular analysis of vertebrates and plants in scats of leopard cats (Prionailurus bengalensis) in southwest China. J. Mammal. 97: 1054–1064. Search in Google Scholar
Xiong, M.Y., D.J. Wang, H.L. Bu, X.N. Shao, D. Zhang, S. Li, R.J. Wang and M. Yao. 2017. Molecular dietary analysis of two sympatric felids in the Mountains of Southwest China biodiversity hotspot and conservation implications. Nat. Sci. Rep. 7: 41909. Search in Google Scholar
Zhang, Y.G., J.E. Janecka, D.Q. Li, H.R. Duo, R. Jackson and W.J. Murphy. 2008. Population survey and genetic diversity of snow leopards (Panthera uncia) as revealed by fecaI DNA. Acta Zool. Sin. 54: 762–766. Search in Google Scholar
The online version of this article offers supplementary material (https://doi.org/10.1515/mammalia-2018- 0144).
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