不同益生菌对肉鸡肠道菌群结构的影响

李可; 罗建杰; 孟昆; 姚斌; 刘国华; 郑爱娟

doi:10.3969/j.issn.1006-267x.2015.11.024

动物营养学报 >

2015 , Vol. 27 >Issue 11: 3516 - 3526

DOI: https://doi.org/10.3969/j.issn.1006-267x.2015.11.024

研究简报 Short Communications

不同益生菌对肉鸡肠道菌群结构的影响

李可 ,
罗建杰 ,
孟昆 ,
姚斌 ,
刘国华 ,
郑爱娟

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1. 中国农业科学院饲料研究所, 农业部饲料生物技术重点开放实验室, 北京 100081;
2. 北京科为博生物科技有限公司, 北京 100086

收稿日期: 2015-05-06

网络出版日期: 2015-11-21

基金资助

国家肉鸡产业技术体系2011-2015年度前瞻性研究(CARS-42-G16);"十二五"科技支撑计划生态环保饲料生产关键技术研发与集成示范(2011BAD26B03)

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Effects of Different Probiotics on Intestinal Microbial Community Structure of Broilers

LI Ke ,
LUO Jianjie ,
MENG Kun ,
YAO Bin ,
LIU Guohua ,
ZHENG Aijuan

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1. Key Laboratory for Feed Biotechnology of the Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China;
2. Beijing Creat-Value Biological Technology Co., Ltd., Beijing 100086, China

Received date: 2015-05-06

Online published: 2015-11-21

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摘要

本试验旨在研究益生菌对肉鸡肠道微生态环境的影响,饲养试验选取1日龄的肉公鸡432只,分成4个组,组1饲喂基础饲粮,组2饲喂基础饲粮+地衣芽孢杆菌(≥4×10⁶ CFU/g饲粮),组3饲喂基础饲粮+屎肠球菌(≥10⁶ CFU/g饲粮),组4饲喂基础饲粮+丁酸梭菌(≥10⁶ CFU/g饲粮)。采用PCR-变性梯度凝胶电泳(DGGE)技术对肉鸡肠道内细菌16S rDNA V3区进行菌群多样性分析,利用实时荧光定量PCR(qPCR)技术对肠道菌群数量进行定量分析。PCR-DGGE指纹图谱结果表明:与对照组相比,地衣芽孢杆菌、屎肠球菌和丁酸梭菌均不同程度的影响了肉鸡回肠、空肠及盲肠样品PCR-DGGE指纹图谱的条带数,其中屎肠球菌处理后回肠、空肠样品PCR-DGGE指纹图谱的条带数显著增加(P<0.05),其余样品PCR-DGGE指纹图谱的条带数无显著变化(P>0.05)。由PCR-DGGE指纹图谱条带明亮度可知:屎肠球菌在回肠食糜中发生了生长繁殖,且促进了乳杆菌生长;地衣芽孢杆菌的添加促进了乳杆菌的生长繁殖,与其产生了协同作用,而抑制了粪肠球菌(C. eutactus)和梭菌(C. irregulare)的生长,二者相互竞争;饲粮中添加丁酸梭菌促进了L. aviaries在空肠及L. versmoldensis在回肠食糜中的生长繁殖,其作用效果比较专一。qPCR结果表明:地衣芽孢杆菌组空肠、回肠及盲肠细菌的菌群数量分别是对照组的1.5倍、1.0倍和1.3倍;屎肠球菌组空肠、回肠及盲肠细菌的菌群数量分别是对照组的4.0倍、6.0倍及0.6倍;丁酸梭菌组空肠、回肠及盲肠细菌的菌群数量分别是对照组的1倍、3.5倍及0.5倍。该研究结果表明地衣芽孢杆菌、屎肠球菌和丁酸梭菌不同程度改变了肉鸡肠道的菌群结构。

关键词： 益生菌; 变性梯度凝胶电泳; 实时荧光定量PCR; 肠道; 菌群结构

本文引用格式

李可 , 罗建杰 , 孟昆 , 姚斌 , 刘国华 , 郑爱娟 . 不同益生菌对肉鸡肠道菌群结构的影响[J]. 动物营养学报, 2015 , 27(11) : 3516 -3526 . DOI: 10.3969/j.issn.1006-267x.2015.11.024

Abstract

This experiment was conducted to study the effects of different probiotics(Bacillus licheniformis, Enterococcus faecium and Clostridium butyricum) on microbial community structure of broilers. A total of 432 one-day-old roosters were randomly allocated to 4 groups, group 1 was fed the basal diet, groups 2 to 4 were fed the basal diet with Bacillus licheniformis (≥4×10⁶ CFU/g), Enterococcus faecium (≥10⁶ CFU/g) and Clostridium butyricum (≥10⁶ CFU/g), respectively. Diversity of intestinal microflora was determined based on V3 variable regions of bacteria 16S rDNA by using PCR-denaturing gradient gel electrophoresis (DGGE) technique. The flora amount of bacteria in the intestine was quantified by real-time quantitative PCR (qPCR). The PCR-DGGE fingerprints results showed as follows:compared with the control group, Enterococcus could significantly increase the number of bands in PCR-DGGE fingerprints (P<0.05), while the number of bands in PCR-DGGE fingerprints of other group was not significantly affected (P<0.05). From the brightness of bands in PCR-DGGE fingerprints, we could know that:Enterococcus faecium was successfully colonized in the ileum, and promote the growth of Lactobacillus, Bacillus licheniformis, also promote the growth of Lactobacillus with a synergistic effect, and inhibit the growth of C. eutactus and C. irregulare with a competition effect. Clostridium butyricum promote the growth of L. aviaries in jejunum and L. versmoldensis in ileum with a more specific effect. The qPCR results showed as follows:the flora amount of bacteria in jejunum, ileum and cecum of Bacillus licheniformis group was 1.5, 1.0 and 1.3 times as many as that of control group; the flora amount of bacteria in jejunum, ileum and cecum of Enterococcus faecium group was 4.0, 6.0 and 0.6 times as many as that of control group; the flora amount of bacteria in jejunum, ileum and cecum of Clostridium butyricum group was 1.0, 3.5 and 0.5 times as many as that of control group. In conclusion, Bacillus licheniformis, Enterococcus faecium and Clostridium butyricum can change the micro-ecology environment of broiler intestine to varying degrees.

Key words： probiotics; denaturing gradient gel electrophoresis; real-time quantitative PCR; intestine; microbial community structure

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