Age-based Compositions And Differences In The Gut Microbiome Of The Shennongjia Golden Snub-nosed Monkey (Rhinopithecus Roxellana Hubeiensis)

The golden snub-nosed monkey as one of the first class national protected animals has a high research value and has been listed in critically endangered species for the rare number. Golden snub-nosed monkeys comprise the genus Rhinopithecus, which include Rhinopithecus roxellana, Rhinopithecus bieti, Rhinopithecus brelichi, Rhinopithecus avunculus, and Rhinopithecus strykeri. Golden snub-nosed monkeys residing within the Shennongjia natural reserve belong to Rhinopithecus roxellana. Numerous of measures have been taken by government in China to protect the golden snub-nosed monkey in Shennongjia. Artificial food supplement plan is one of them. The implementation of the artificial food supplement plan has changed the original living environment and the diet structure of the golden snub-nosed monkey and then changed compositions of the intestinal flora. The intestinal flora is closely related to the intestinal health and the imbalance of the intestinal flora may result in intestinal diseases and non-intestinal diseases. Therefore, the research on the intestinal flora of different-aged golden snub-nosed monkeys has great significance in monitoring the intestinal diseases and it also provides important theoretical and practical significance to the artificial diet development of the golden snub-nosed monkey.Here we characterized the bacterial species in fecal samples from 47 golden snub-nosed monkeys in Shennongjia, Hubei Province. 17 random samples of them were used in denaturing gradient gel electrophoresis(DGGE) of PCR-amplified fragments to explore the diversity and composition of the gut microbiota; The other 30 fecal samples collecting from 30 golden snub-nosed monkeys were used for high throughput sequencing. The bacterial 16 S r RNA gene V4 region was amplified and then sequenced on the Illumina Mi Seq platform PE250 to identify the bacterial constituents and differences. The result of the intestinal flora from high throughput sequencing had also been compared with the other species. The results and the conclusions display as following. 1. The results of the 17 random fecal samples of denaturing gradient gel electrophoresis indicated that: Different-aged golden snub-nosed monkeys had differences in gut microbiota’s composition; Firmicutes(55.6%)and Bacteroidetes(13.9%) were the main constitution in golden snub-nosed monkey’s gut. 2. The results of the 30 different-aged fecal samples of high-throughput sequencing indicated that: The main phyla of the golden snub-nosed monkey’s gut and the corresponding proportion of the main phyla were Firmicutes(20.96%-30.29%) and Bacteroidetes(29.85%-39.46%); The operational taxonomic units(OTUs) numbers in 0-3 years-old group(Y1), 5-8 years-old group(Y2), above 12 years-old group(Y3) were 1469, 1381, 1931 respectively; The plyla numbers in 0-3 years-old group(Y1), 5-8 years-old group(Y2), above 12 years-old group(Y3) were 16, 16, 19 and genus numbers were 166, 157, 190 respectively; The OUTs numbers in different-aged golden snub-nosed monkeys indicated that the species number of the intestinal flora in above 12 years-old group(Y3) was the most abundant group; UPGMA cluster analysis showed that the species composition of gut microbiota in 0-3 years-old group(Y1) and 5-8 years-old group(Y2) had a high similarity, however, had a distance with above 12 years-old group(Y3). 3. Compared with the other species, the results showed: Firmicutes and Bacteroidetes were preponderant phyla in the gut of human, non-human primates and the golden snub-nosed monkey, but the proportion of Firmicutes and Bacteroidetes in the three species was different(the golden snub-nosed monkey: 57.88%-62.17%; human: above 90%; non-human primates: 70.50%-98.30); The proportion of Proteobacteria had a considerable difference in the gut of human and golden snub-nosed monkeys; Spirochaetes was a dominant phylum in the gut of golden snub-nosed monkeys, but was not in the gut of human and non-human primates. In the gut of the golden snub-nosed monkey, Clostridia was the main class of the phylum Firmicutes; Fibrobacteres as a typical phylum of ruminant existed in the golden snub-nosed monkey’s gut; Methanobrevibacter as a typical bacterium of ruminant existed in the gut of the golden snub-nosed monkey.