基于16S rRNA测序分析正常与腹泻牦牛犊牛粪便细菌区系组成

doi:10.16431/j.cnki.1671-7236.2021.09.020

摘要/Abstract

摘要： 试验旨在比较分析放牧条件下正常与腹泻牦牛犊牛粪便微生物区系组成与基因功能预测，筛选出作为腹泻评价依据的细菌，为下一步牦牛犊牛益生菌制剂的开发提供理论依据。在放牧条件下分别采集青海玉树地区正常与腹泻牦牛犊牛粪便样本各7份并提取DNA，利用16S rRNA测序技术研究两组牦牛犊牛肠道微生物区系的OTUs聚类、门水平和属水平相对丰度差异性、Alpha多样性、Beta多样性、功能预测水平信息。结果显示，在门水平上，正常组和腹泻组的优势菌门有拟杆菌门和厚壁菌门，且腹泻组的梭杆菌门显著高于正常组（P<0.05）；在属水平上，正常组犊牛粪便中优势细菌相对丰度从高到低为：拟杆菌属、拟普雷沃菌属和未分类的瘤胃菌属；腹泻组犊牛粪便中优势细菌相对丰度从高到低为：拟杆菌属、梭杆菌属、未分类的肠杆菌属和未分类的瘤胃菌属，且腹泻组的梭杆菌属显著高于正常组（P<0.05）；在功能预测水平上，腹泻组的聚糖生物合成与代谢、运输和分解代谢、老化、分类和降解、脂代谢显著高于正常组（P<0.05），正常组的氨基酸代谢、信号转导、细胞运动、遗传信息显著高于腹泻组（P<0.05）。通过分析放牧牦牛犊牛正常组和腹泻组之间的优势菌群差异表明，犊牛腹泻是由梭杆菌属引起的。

关键词: 牦牛犊牛; 腹泻; 粪便; 细菌区系; 16S rRNA测序

Abstract: This experiment was conducted to compare and analyze the fecal microflora composition and gene function prediction of normal and diarrhea yak calves under grazing conditions, and screen out the bacteria as the basis for diarrhea evaluation, so as to provide the theoretical basis for the development of probiotics for yak calves in the next step. Seven fecal samples were collected from normal and diarrhea yak calves in Yushu area of Qinghai province under grazing conditions, respectively, and DNA was extracted. 16S rRNA sequencing technology was used to study the OTU clustering, relative abundance difference at phylum level and genus level, Alpha diversity, Beta diversity and functional prediction level information of intestinal microflora of yak calves between normal and diarrhea groups. The results showed that at phylum level, Bacteroidetes and Firmicutes were the dominant phyla in normal and diarrhea groups, and the Fusobacterium in diarrhea group was significantly higher than that in normal group (P<0.05). At genus level, the relative abundance of dominant bacteria in the feces of normal group calves from high to low were Bacteroides, Alloprevotella and unidentified_Ruminococcaceae. The relative abundance of dominant bacteria in the feces of diarrhea group calves from high to low were Bacteroides, Fusobacterium, unidentified_Enterobacteriaceae and unidentified_Ruminococcaceae, and the abundance of Fusobacterium in diarrhea group was significantly higher than that in normal group (P<0.05). At function prediction level, the main contents of glycan biosynthesis and metabolism, transportation and catabolism, aging, classification and degradation and lipid metabolism in diarrhea group were significantly higher than that in normal group (P<0.05), while the main contents of amino acid metabolism, signal transduction, cell movement and genetic information in normal group were significantly higher than that in diarrhea group (P<0.05). According to this experiment, the difference of dominant flora between normal and diarrhea groups of grazing yak calves was analyzed, the diarrhea of calves in this experiment was caused by Fusobacterium.

Key words: yak calves; diarrhea; fecal; bacterial flora; 16S rRNA sequencing

中图分类号:

S823.8⁺5

张玉莹, 刘书杰, 冯宇哲, 张晓卫, 崔占鸿. 基于16S rRNA测序分析正常与腹泻牦牛犊牛粪便细菌区系组成[J]. 中国畜牧兽医, 2021, 48(9): 3293-3302.

ZHANG Yuying, LIU Shujie, FENG Yuzhe, ZHANG Xiaowei, CUI Zhanhong. Bacterial Flora Composition of Normal and Diarrhea Yak Calves Based on 16S rRNA Sequencing[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(9): 3293-3302.

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