Peer Review #2 of "Comparison of the fecal microbiota of two free-ranging Chinese subspecies of the leopard (Panthera pardus) using high-throughput sequencing (v0.1)"

Abstract

The analysis of gut microbiota using fecal samples provides a non-invasive approach to understand the complex interactions between host species and their intestinal bacterial community. However, information on gut microbiota for wild endangered carnivores is scarce. The goal of this study was to describe the gut microbiota of two leopard subspecies, the Amur leopard (Panthera pardus orientalis) and North Chinese leopard (Panthera pardus japonensis). Fecal samples from the Amur leopard (n=8) and North Chinese leopard (n=13) were collected in Northeast China Tiger and Leopard National Park and Shanxi Tieqiaoshan Provincial Nature Reserve, respectively. The gut microbiota of leopards was analyzed via high-throughput sequencing of the V3-V4 region of bacterial 16S rRNA gene using the Life Ion S5TM XL platform. A total of 1,413,825 clean reads representing 4,203 operational taxonomical units (OTUs) were detected. For Amur leopard samples, Firmicutes (78.4%) was the dominant phylum, followed by Proteobacteria (9.6%) and Actinobacteria (7.6%). And for North Chinese leopard, Firmicutes (68.6%), Actinobacteria (11.6%) and Proteobacteria (6.2%) were the most predominant phyla. Clostridiales was the most diverse bacterial order with 37.9% for Amur leopard and 45.7% for North Chinese leopard. Based on the beta-diversity analysis, no significant difference was found in the bacterial community composition between the Amur leopard and North Chinese leopard samples. The current study provides the initial data about the composition and structure of the gut microbiota for wild Amur leopards and North Chinese leopards, and has laid the foundation for further investigations of the health, dietary preferences and physiological regulation of leopards.