The Study On The Bacterial And Archaeal Diversity In The Feces Of The Gaint Padans And The Forest Musk Deers

In this study, five healthy adult captive giant pandas and five healthy forest musk deers were choosed as experimental animals. Their fresh feces before the morning feeding were collected to evaluate their bacterial and archaeal compositions using high-throughput sequencing technology.The total microbial DNA was extracted from fecal samples of each panda and forest musk deer,then the DNA was used as template to amplify the V3-V4 variable region with length about 300bp of 16S rRNA gene using a bacteria-specific primer pair 515F (5’-GTG CCAGCMGCCGCGGTAA-3’),806R(5’-GGACTACVSGGGTATCTAAT-3’).The DNA was also used as template to amplify the V4-V5 variable region with the length about 500bp of 16S rRNA gene using a archaea-specific primer pair 349F (5’-GYGCASCAGKC GMGAAW-3’),806R(5’-GGACTACVSGGGTATCT AAT-3’). The PCR products were visualized by 2% agarose gel electrophoresis and purified using a PCR purification kit.The amplification of bacteria was subjected to sequencing on Illumina’s MiSeq 250PE sequencing platforms (Megiddo biological) and the amplification of archaea on Illumina’s MiSeq 300PE sequencing platforms (Millennium Gene). The obtained sequences were undergone quality control and filtering, clustering analysis, species taxonomic analysis, and the ultimate results were following:The bactetial in the feces of giant pandas and forest musk deers were assigned to 16 plyla and 21 plyla,respectively,and 13 phyla were shared by all animal fecal samples. At the phylum level,the dominant bacteria were Proteobacteria (74.46%) and Firmicutes (15.66%) in giant pandas’feces,and Firmicutes (51.53%) and Bacteroidetes (34.69%) in forest musk deers’feces. At the genus level, the predominant bacteria were Escherichia/Shigella (49.84%) and Enterobacteriaceae (Other)(13.73%) in giant pandas’feces.and Ruminococcaceae (other)(15.10%) and Clostridiales(other) (14.17%) in forest musk deers’ feces.The difference of this four genera was extremely significant (P<0.01) between in the feces of giant pandas and forest musk deers.In giant pandas’feces,the abundance of phylum Crenarchaeota was higher than phylum Euryarchaeota.At the genus level, the dominant archaea were Thermoplasmatales (unclassified) (57.91%) and Methanobrevibacter (18.35%) in giant pandas’feces,and Methanobrevibacter (73.18%) and Thermoplasmatales (unclassified) (12.01%)in Forest musk deers’feces. The abundance of Methanobrevibacter and Methanosphaera in the feces of giant pandas and forest musk deer were significant difference (P<0.05).