Comparative The Fecal Flora Of Health And Diarrhea Adult Rhinopithecus Roxellanae

This study aim to explore the differences of intestinal flora between diarrheal and healthy Rhinopithecus roxellanae and to protect the Rhinopithecus roxellanae better. Test collected each 15 healthy and diarrheal feces of adult Rhinopithecus roxellanae at the zoo of Chengdu in March tu April,2014 (7 feces of each group collected by March compared preliminarily by PCR-DGGE analysis). These 30 samples were analyzed for high-throughput sequencing by using Illumina technology and by Real-time PCR on the total number of bacteria as below:Clostridium cluster IV, Clostridium cluster ⅪⅤa, Bacteroides prevotella, Lactobacillus spp. and Enterobacteriaceae. The test results were as follows:1. The PCR-DGGE profiles showed there had no significant difference with microbial diversity index (healthy group 3.87±0.02, diarrheal group 3.87±0.01) and richness (healthy group 48±2.16, diarrheal group 48±2.45) in diarrheal group comparing with the healthy group. The two groups separated obviously to two part in cluster analysis. The similarity between the two groups was 0.78. The similarity between healthy samples was 0.81 to 0.86 and the similarity between diarrheal samples was 0.83 to 0.92.2. Results of 16s rRNA high-throughput sequencing:Average per sample detected 18912±6527 tags in all the samples. There had no significant difference between healthy group (19172±6806) and diarrheal group (18652±6463) among all the samples. OTU of samples was calculated at the similarity level of 97% by using Mothur had no significant between healthy group (2598±971) and diarrheal group (2367±825). On the level of phylum, the main floras of healthy adult Rhinopithecus roxellanae were Firmicutes, Bacteroidetes. There had significant changes with five kinds of Phylum, seven kinds of Class, eight kinds of Order,19 kinds of Family and 28 kinds of Genus between diarrheal group and healthy group. On the level of Genus, Flexispira was average of 3.43×10-04 and the total of detection rate was 90% (healthy group 13/15, diarrheal group 14/15).3. Results of Real-time PCR:The number of Total Bacteria was 5.14x1011cfu/g in feces flora of healthy Rhinopithecus roxellanae. Clostridium cluster IV and Clostridium cluster ⅪⅤa were total to 2.11×1011 cfu/g which was 41% of the total number of bacteria. Comparing diarrheal group and healthy group, there had no significant difference in Total Bacteria, Clostridium cluster IV and Clostridium cluster ⅪⅤa. Bacteroides prevotella had the trend of decrease but had no obvious difference. Enterobacteriaceae rised observably but Lactobacillus spp. reduced observably.In conclusion, the feces flora of adult Rhinopithecus roxellanae is variety, and the Firmicutes and Bacteroides were the dominant species. Compared with the normal group the differences of the total number an diversity of feces flora of the diarrhea group have no significant, but there had significant differences on some microflora, compared to the normal group the lactobacillus, Prevotella was significantly decreased, while the number of some pathogens flora (Facklamia, Erysipelothrix, Acinetobacter) were significantly increased, which was affect the health of Rhinopithecus roxellanae.