Efficient and cost-effective non-invasive population monitoring as a method to assess the genetic diversity of the last remaining population of Amur leopard (Panthera pardus orientalis) in the Russia Far East

PLoS One. 2022 Jul 6;17(7):e0270217. doi: 10.1371/journal.pone.0270217. eCollection 2022.

Abstract

Small populations of the endangered species are more vulnerable to extinction and hence require periodic genetic monitoring to establish and revisit the conservation strategies. The Amur leopard is critically endangered with about 100 individuals in the wild. In this study, we developed a simple and cost-effective noninvasive genetic monitoring protocol for Amur leopards. Also, we investigated the impact of fecal sample's age, storage, and collection season on microsatellite genotyping success and data quality. We identified 89 leopard scats out of the 342 fecal samples collected from Land of the Leopard between 2014-2019. Microsatellite genotyping using 12 markers optimized in 3 multiplex PCR reactions reveals presence of at least 24 leopard individuals (18 males and 6 females). There was a significant difference in the success rate of genotyping depending on the time from feces deposition to collection (p = 0.014, Fisher's exact test), with better genotyping success for samples having <2 weeks of environmental exposure. Amur leopard genetic diversity was found low (Ho- 0.33, HE- 0.35, and NA- 2.57) with no visible population substructure and recent bottleneck signature. Although a historical bottleneck footprint was observed. Mitochondrial DNA diversity was also found low with two haplotypes differing by a point mutation reported in 1,769 bp of investigated sequence covering parts of cytochrome b gene (846 bp), NADH-5 gene (611 bp) and control region (312 bp). We recommend periodic genetic monitoring of wild Amur leopards following the proposed methodology to achieve cost effectiveness and efficiency.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Asia, Eastern
  • Cost-Benefit Analysis
  • Endangered Species
  • Female
  • Genetic Variation
  • Male
  • Panthera* / genetics

Grants and funding

This work was supported by the following. Ministry of Environment of the Republic of Korea, Convention on Biological Diversity under Bio-Bridge Initiative (550-20190003): SC, PP, JYH, DJ, JL, DYK, M-SM, HL, Brain Korea-21 programme (5260-20190100, 5260-20200100, A0449-20200100): SC, DJ, JL, Brain Korea-21 programme (A0449-20210100): SC. The funders had no role in study design, data collection, and analysis, decision to publish, or preparation of the manuscript.