一组微卫星标记，用于对个体豹进行识别，及其在人类活动强烈干扰的景观中对豹监测的应用。
A panel of microsatellites to individually identify leopards and its application to leopard monitoring in human dominated landscapes.
机构信息
National Centre for Biological Sciences, TIFR, GKVK Campus, Bellary Road, Bangalore 560065, India.
出版信息
BMC Genet. 2009 Dec 4;10:79. doi: 10.1186/1471-2156-10-79.
BACKGROUND
Leopards are the most widely distributed of the large cats, ranging from Africa to the Russian Far East. Because of habitat fragmentation, high human population densities and the inherent adaptability of this species, they now occupy landscapes close to human settlements. As a result, they are the most common species involved in human wildlife conflict in India, necessitating their monitoring. However, their elusive nature makes such monitoring difficult. Recent advances in DNA methods along with non-invasive sampling techniques can be used to monitor populations and individuals across large landscapes including human dominated ones. In this paper, we describe a DNA-based method for leopard individual identification where we used fecal DNA samples to obtain genetic material. Further, we apply our methods to non-invasive samples collected in a human-dominated landscape to estimate the minimum number of leopards in this human-leopard conflict area in Western India.
RESULTS
In this study, 25 of the 29 tested cross-specific microsatellite markers showed positive amplification in 37 wild-caught leopards. These loci revealed varied levels of polymorphism (four-12 alleles) and heterozygosity (0.05-0.79). Combining data on amplification success (including non-invasive samples) and locus specific polymorphisms, we showed that eight loci provide a sibling probability of identity of 0.0005, suggesting that this panel can be used to discriminate individuals in the wild. When this microsatellite panel was applied to fecal samples collected from a human-dominated landscape, we identified 7 individuals, with a sibling probability of identity of 0.001. Amplification success of field collected scats was up to 72%, and genotype error ranged from 0-7.4%.
CONCLUSION
Our results demonstrated that the selected panel of eight microsatellite loci can conclusively identify leopards from various kinds of biological samples. Our methods can be used to monitor leopards over small and large landscapes to assess population trends, as well as could be tested for population assignment in forensic applications.
背景
豹是大型猫科动物中分布最广的物种，从非洲到俄罗斯远东地区都有它们的身影。由于栖息地的碎片化、高人口密度以及该物种固有的适应性，它们现在占据了靠近人类住区的景观。因此，它们是印度最常见的涉及人与野生动物冲突的物种，需要对其进行监测。然而，豹行踪隐秘，这使得监测工作变得困难。近年来，DNA 方法的进步以及非侵入性采样技术的发展，可以用来监测包括人类主导景观在内的大片区域的种群和个体。在本文中，我们描述了一种基于 DNA 的豹个体识别方法，该方法使用粪便 DNA 样本来获取遗传物质。此外，我们将我们的方法应用于在人类主导的景观中收集的非侵入性样本，以估计印度西部这个人与豹冲突地区的豹数量。
结果
在这项研究中，29 个交叉特异性微卫星标记中的 25 个在 37 只野生捕获的豹中显示出阳性扩增。这些位点显示出不同程度的多态性（4-12 个等位基因）和杂合度（0.05-0.79）。结合扩增成功率（包括非侵入性样本）和位点特异性多态性的数据，我们表明，8 个位点的兄弟姐妹概率为 0.0005，这表明该面板可用于识别野生个体。当将这个微卫星面板应用于从人类主导的景观中收集的粪便样本时，我们鉴定出了 7 个个体，其兄弟姐妹概率为 0.001。现场采集的粪便样本的扩增成功率高达 72%，基因型错误率为 0-7.4%。
结论
我们的结果表明，从各种生物样本中可以使用所选的 8 个微卫星位点的面板来明确识别豹。我们的方法可用于监测小型和大型景观中的豹，以评估种群趋势，也可以在法医应用中进行种群分配的测试。
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