Data from: Carnivore diet analysis based on next-generation sequencing: application to the leopard cat (Prionailurus bengalensis) in Pakistan

Diet analysis is a prerequisite to fully understand the biology of a species and the functioning of ecosystems. For carnivores, traditional diet analyses mostly rely upon the morphological identification of undigested remains in the feces. Here, we developed a methodology for carnivore diet analyses based on next generation sequencing. We applied this approach to the analysis of the vertebrate component of leopard cat diet in two ecologically distinct regions in northern Pakistan. Despite being a relatively common species with a wide distribution in Asia, little is known about this elusive predator. We analyzed a total of 38 leopard cat feces. After a classical DNA extraction, the DNA extracts were amplified using primers for vertebrates targeting about 100 bp of the mitochondrial 12S rRNA gene, with and without a blocking oligonucleotide specific to the predator sequence. The amplification products were then sequenced on a next generation sequencer. We identified a total of 18 prey taxa, including eight mammals, eight birds, one amphibian, and one fish. In general, our results confirmed that the leopard cat has a very eclectic diet, and feeds mainly on rodents, and particularly on the Muridae family. The DNA-based approach we propose here represents a valuable complement to current conventional methods. It can be applied to other carnivore species with only a slight adjustment relating to the design of the blocking oligonucleotide. It is robust, simple to implement, and allows the possibility of very large-scale analyses.饮食分析是全面理解某一物种生物学特性与生态系统运作机制的先决条件。对于食肉动物而言，传统饮食分析大多依赖于对粪便中未消化残余物的形态学鉴定。本研究开发了一种基于下一代测序（next generation sequencing）的食肉动物饮食分析方法，并将其应用于巴基斯坦北部两个生态迥异区域中豹猫（leopard cat）饮食的脊椎动物组分分析。尽管该物种在亚洲分布广泛且相对常见，但人们对这种神秘莫测的食肉动物所知甚少。本研究共分析了38份豹猫粪便样本。在进行经典DNA提取后，我们使用靶向线粒体12S核糖体RNA（mitochondrial 12S rRNA）基因约100 bp片段的脊椎动物引物，分别添加和不添加针对该食肉动物序列的封闭寡核苷酸，对DNA提取物进行扩增。扩增产物随后在下一代测序仪上完成测序。本研究共鉴定出18个猎物类群，其中包括8种哺乳类、8种鸟类、1种两栖类和1种鱼类。总体而言，研究结果证实豹猫饮食极为宽泛，主要以啮齿类动物为食，尤其是鼠科（Muridae）物种。本研究提出的基于DNA的分析方法，可作为现有传统方法的宝贵补充手段。仅需对封闭寡核苷酸的设计进行小幅调整，该方法即可应用于其他食肉动物物种的相关研究。该方法稳定性强、操作简便，且可支持大规模分析研究。