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<div>1Northeast Tiger and Leopard Biodiversity National Observation and Research Station, Ministry of Education Key Laboratory for Biodiversity Science and Ecological Engineering, State Forestry and Grassland Administration Key Laboratory for Conservation Ecology of Northeast Tiger and Leopard National Park, State Forestry and Grassland Administration Amur tiger and Amur leopard Monitoring and Research Center, College of Life Science, Beijing Normal University, Beijing, China</div><div></div><div></div><div>Amur Leopard and North Chinese leopard are large-sized feline species and solitary predators that play pivotal roles in the ecosystems where they occur. Many efforts have been made to uncover their dietary habits, population genetic structure, and individual identification for conservation purposes through non-invasive sampling of feces (Dutta & Seidensticker, 2013; Dutta et al., 2012; Rodgers & Janečka, 2013; Yang et al., 2018). Gut microbial diversity analyses based on leopard fecal samples should also be considered as an important part of conservation efforts. In-depth understanding of the relationship between host habitat and microbiota composition may be helpful for conservation efforts because changes in the gut bacterial communities have been shown to affect host metabolism and energy homeostasis (Musso, Gambino & Cassader, 2010).</div><div></div><div></div><div></div><div></div><div></div><div>mac os leopard download kostenlos</div><div></div><div>DOWNLOAD: https://t.co/vSUUXNnA67 </div><div></div><div></div><div>Series of alpha-diversity indices including Observed species, Shannon, Simpson, Chao1, ACE, and Goods coverage were calculated and analyzed in QIIME (Version 1.9.1) (Caporaso et al., 2010). The rarefaction curves and rank abundance curves were constructed in R (Version 2.15.3). We applied Wilcoxon rank-sum test to identify discrepancies of gut bacterial diversities between the Amur leopard and North Chinese leopard for each index of alpha-diversity.</div><div></div><div></div><div>For the Amur leopard, Firmicutes was the predominant phylum (78.4%) (Fig. 2). Proteobacteria (9.6%), Actinobacteria (7.6%), Bacteroidetes (2.6%) and Fusobacteria (1.7%) contributed also to the total composition. At the family level, Planococcaceae (30.1%), Clostridiaceae 1 (17.2%) and Peptostreptococcaceae (14.5%) were the top 3 dominant families. At the genus level, Sporosarcina was predominant with an abundance of 22.8%, followed by Clostridium sensu stricto 1 (17.1%) and Peptoclostridium (10.2%).</div><div></div><div></div><div>For the North Chinese leopard, Firmicutes (68.6%) was the most predominant phylum (Fig. 2), followed by Actinobacteria (11.6%), Fusobacteria (6.4%), Proteobacteria (6.2%) and _Bacteroidetes _(6.0%). Clostridiaceae_1 (19.5%), Planococcaceae (16.2%) and Lachnospiraceae (12.5%) were the three most predominant families. At the genus level, Clostridium sensu stricto 1 (19.4%), Sporosarcina (9.5%) and Peptoclostridium (6.1%) constituted the top three genera.</div><div></div><div></div><div>The yellow points represent Amur leopard and the blue squares represent North Chinese leopard. For PCoA (C) and (D) were analyzed with weighted Unifrac distance and unweighted Unifrac distance respectively. All the points are scattered, which indicates that no significant differences were found between the two subspecies.</div><div></div><div></div><div>Five major bacterial phyla were observed including Firmicutes, Proteobacteria, Actinobacteria, Bacteroidetes and Fusobacteria both in the Amur leopard and North Chinese leopard samples, which is in accordance with the vertebrate gut microbial diversity described by many other studies (Deng & Swanson, 2014; Ley et al., 2008; Ritchie, Steiner & Suchodolski, 2008). Fecal samples of healthy cats are featured with similar phylum composition with slightly different proportions (Barry et al., 2012). Based on our analysis, no significant difference was found in the relative abundance of these five phyla between the samples from the Amur leopard and North Chinese leopard.</div><div></div><div></div><div>Firmicutes was the most predominant phylum in both the Amur leopard and North Chinese leopard and showed no significant difference between two groups (p = 0.210). Previous researches have reported that Firmicutes is the most dominant phylum in feces of animals (Garcia-Mazcorro et al., 2012; Guan et al., 2017; Ritchie et al., 2010) and humans (Arumugam et al., 2011). Same tendency was also found in feline species in the wild such as leopard cats (Prionailurus bengalensis) (An et al., 2017) and snow leopards (Panthera uncia) (Zhang et al., 2015). Some studies reported that the body fat storage influences the gut bacterial composition in mice (Ley et al., 2005) and humans (Ley, Peterson & Gordon, 2006). A significantly greater proportion of Firmicutes and a significant reduction of Bacteroidetes were observed in obese animals than in lean controls (Turnbaugh et al., 2006). The tendency of an increase in Firmicutes and a decrease in Bacteroidetes was associated with switching to the high fat diet (Hildebrandt et al., 2009; Tremaroli & Bäckhed, 2012). We detected that the proportion of Firmicutes in Amur leopards was relatively greater than in North Chinese leopards, and the proportion of Bacteroidetes in Amur leopards was relatively lower which indicated that the weight of Amur leopard should be more heavier. This might relate to the greater body fat storage of Amur leopards compare with North Chinese leopards, since Amur leopards have larger body size and store more fat to withstand severe cold in further north habitat (Wang et al., 2017). Unfortunately, the detail information about wild North Chinese leopard is comparatively scarce.</div><div></div><div></div><div></div><div></div><div></div><div></div><div>Within the phylum Firmicutes, Zhang et al. (2015) found that Lachnospiraceae was the most diverse family in the feces of snow leopards, which is consistent with a previous report in wolves (Canis lupus) (Zhang & Chen, 2010). In our results, however, the most diverse family was Clostridiaceae 1 (19.5% in North Chinese leopard, 17.2% in Amur leopard) within the order Clostridiales, and Lachnospiraceae constituted a relatively small proportion in our sample set compared to snow leopards and wolves. Lachnospiraceae was found in both human and mammal gut microbiota that relates to some diseases like colon cancer (Meehan & Beiko, 2014), nonalcoholic fatty liver disease (NAFLD) (Shen et al., 2017) and diabetes (Kameyama & Itoh, 2014). However, without sufficient support based on other health monitoring methods including blood or apparatus test, the proportion of Lachnospiraceae in the gut microbiota could only be a simple referential marker that reflects health condition for wild animals.</div><div></div><div></div><div>Our results also indicated that Clostridium sensu stricto 1 was a predominant genus in the gut microbiota of leopards. And Clostridium perfringens was a common bacterial species for both the Amur leopard and North Chinese leopard. Lubbs et al. (2009) reported that the gut microbiota of domestic cats is affected by the protein concentration in diets, particularly, Clostridium populations increased as more protein was digested. The presence of C. perfringens was positively associated with protein intake in grizzly bears (Ursus arctos) (Schwab et al., 2011) and cheetahs (Acinonyx jubatus) (Becker et al., 2014). To our knowledge, leopards are highly carnivorous and consume mostly protein in their daily diet (Martins et al., 2011). We speculate that the high proportions of Clostridium populations might reflect the high-protein diet of leopards in our study. Interestingly, C. perfringens might be potential pathogenic bacteria that cause diarrhea in dogs (Canis lupus familiaris) and cats (Felis catus) (Suchodolski, 2011). However, C. perfringens was also detected in the clinically healthy dogs and house cats (Handl et al., 2011; Queen, Marks & Farver, 2012). C. perfringens should probably be considered as a common commensal in the intestine of healthy feline (Becker et al., 2014).</div><div></div><div> df19127ead</div>