3～6月龄伊犁马肠道微生物群落多样性的研究

A Study on Intestinal Microbiota Diversity of 3-to 6-Month-Old Yili Horses

本试验旨在研究3~6月龄伊犁马肠道微生物群落的多样性,揭示断奶前马驹肠道菌群演替的规律,从微生物学角度为此阶段马驹生长发育及肠道健康研究提供理论依据。试验选取平均体重为(89.75±8.81)kg、出生日期相同的3月龄伊犁马5匹,进行为期90 d的饲养试验。分别在试验的第0天(即试验开始的前1天)、第30天、第60天、第90天采集马驹粪便样品,从每份样品中提取微生物基因组总DNA,采用Illumina HiSeq测序技术检测样品微生物群落多样性。结果显示:1)对5匹马驹的20份粪样进行测序,共获得有效序列数157 665条,获得平均操作分类单元(OTUs)1 117个。2)马驹粪便中微生物群落α多样性指数(ACE、Chao1、Shannon和Simpson指数)随着马驹月龄的增加呈波动性变化,但各时间点之间均无显著性变化(P>0.05)。3)在门水平上,马驹粪便中10大优势菌分别为厚壁菌门(Firmicutes)、拟杆菌门(Bacteroidetes)、变形菌门(Proteobacteria)、疣微菌门(Verrucomicrobia)、螺旋体菌门(Spirochaetes)、梭杆菌门(Fusobacteria)、无壁菌门(Tenericutes)、放线菌门(Actinobacteria)、TM7、广古菌门(Euryarchaeota),其中以厚壁菌门、拟杆菌门、变形菌门及疣微菌门的丰度较高;在科水平上,马驹粪便中10大优势菌分别为芽孢杆菌科(Bacillaceae)、莫拉氏菌科(Moraxellaceae)、动球菌科(Planococcaceae)、肉杆菌科(Carnobacteriaceae)、BS11、RFP12、乳杆菌科(Lactobacillaceae)、瘤胃球菌科(Ruminococcaceae)、毛螺旋菌科(Lachnospiraceae)、紫单胞菌科(Porphyromonadaceae);在属水平上,马驹粪便中10大优势菌分别为不动杆菌属(Acinetobacter)、德库菌属(Desemzia)、乳杆菌属(Lactobacillus)、Ureibacillus、Paludibacter、芽孢杆菌属(Bacillus)、埃希式杆菌属(Escherichia)、肉食杆菌属(Carnobacterium)、密螺旋体属(Treponema)、艰难杆菌属(Mogibacterium)。由此得出,采用Illumina HiSeq测序技术能够准确地对3~6月龄伊犁马肠道微生物群落进行分类研究;厚壁菌门、拟杆菌门、变形菌门、疣微菌门是3~6月龄伊犁马肠道内的优势菌门。

The current study was conducted to evaluate the intestinal microbiota diversity of 3- to 6-month-old Yili horses, to reveal the regulation of succession of intestinal micrbiota before weaning, and to provide a theo- retical basis for the researches of growth and intestinal health of young horse. Five 3-month-old Yili horses with the average body weight of （ 89.75_＋8.81 ） kg and the same birth day were selected to conduct a 90-day feeding trial. The feces samples were collected on days 0 （ 1 day before the experiment）, 30, 60 and 90 of the experi- ment, respectively. The microbial genomic total DNA was extracted from each feces sample. Microbiota diver- sity of feces was evaluated using lllumina HiSeq sequencing technology. The results showed as follows ： 1 ） twenty feces samples for 5 colts were sequenced, a total of 157 665 effective sequences and 1 117 average op- erational taxonomic units （OTUs） were acquired. 2） The alpha diversity indexes （ACE, Chaol, Shannon and Simpson indices） of feces microbiota showed fluctuated changes with the age increasing, but the changes were not significant among different time points （P〉0.05）. 3） At the phylum level, ten dominant bacteria in feces of colts were Firmicutes, Bacteroidetes, Proteobacteria, Verrucomicrobia, Spirochaetes, Fusobacteria, Teneri- cutes, Actinobacteria, TM7 and Euryarchaeota, and the abundances of Firmicutes, Bacteroidetes, Proteobacte- ria and Verrucomicrobia were higher. At the family level, ten dominant bacteria in feces of colts were Bacil- laceae, Moraxellaceae, Planococcaceae, Carnobacteriaceae, BSll, RFP12, Lactobacillaceae, Ruminococ- caceae, Lachnospiraceae and Porphyromonadaceae. At the genera level, ten dominant bacteria in feces of colts were Acinetobacter, Desemzia, Lactobacillus, Ureibacillus, Paludibacter, Bacillus, Escherichia, Carnobac- terium, Treponema and Mogibacterium. Base on the results, it is concluded that the intestinal microbiota of 3- to 6-month-old Yili horses can be accurately classified using Illumina HiSeq sequencing technology. The Firmi- cutes. Bacteroidetes. Proteobacteria. Verrucomicrobia were the dominant bacteria in intestine of 3- to 6-month-old Yili horses.