猪源益生芽孢杆菌的分离鉴定与生物学特性分析

蒋佳璇; 王淑京; 任志鸿; 杜华茂

doi:10.3969/j.issn.1006-267x.2016.11.022

动物营养学报 >

2016 , Vol. 28 >Issue 11: 3542 - 3548

DOI: https://doi.org/10.3969/j.issn.1006-267x.2016.11.022

饲料营养 Feed Science and Technology

猪源益生芽孢杆菌的分离鉴定与生物学特性分析

蒋佳璇 ,
王淑京 ,
任志鸿 ,
杜华茂

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1. 西南大学生物技术学院, 重庆 400715;
2. 中国疾病预防控制中心传染病预防控制所, 传染病预防控制国家重点实验室, 北京 102206

收稿日期: 2016-04-09

网络出版日期: 2016-11-18

基金资助

传染病预防控制国家重点实验室留学归国人员启动经费（2011SKLlD301）

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Isolation, Identification and Biological Characterization of Bacillus spp from Swine

JIANG Jiaxuan ,
WANG Shujing ,
REN Zhihong ,
DU Huamao

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1. College of Biotechnology, Southwest University, Chongqing 400715, China;
2. National Institute for Communicable Diseases Control and Prevention Chinese Center for Disease Control and Prevention, State Key Laboratory for Infectious Disease Prevention and Control, Beijing 102206, China

Received date: 2016-04-09

Online published: 2016-11-18

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摘要

本研究旨在从健康猪肠道正常菌群中筛选出性能良好的芽孢杆菌，以用于提高饲料利用率、减少猪消化道感染及减少感染后抗生素的使用。本研究精选重庆偏远山区从未添食配合饲料且生产性能优良的经产母猪，从其新鲜粪便中分离耐酸、耐胆盐的芽孢杆菌，进一步筛选能同时产淀粉酶、纤维素酶和蛋白酶的菌株，最后进行生化鉴定和基于16S rDNA序列的分子鉴定。结果表明，试验共筛选出7株能同时分泌淀粉酶、纤维素酶、蛋白酶的芽孢杆菌，它们在pH 3.0条件下处理2 h存活率为30%~90%，在0.5%胆盐中处理2 h存活率为40%~100%。经生化鉴定与分子鉴定为1株嗜热脂肪芽孢杆菌，1株地衣芽孢杆菌，5株枯草芽孢杆菌。由结果可知，本研究筛选的7株芽孢杆菌能耐受低pH和高胆盐环境，且具备同时分泌淀粉酶、纤维素酶和蛋白酶3种胞外酶的能力。

关键词： 益生菌; 芽孢杆菌; 生化鉴定; 16S rDNA; 产酶能力

本文引用格式

蒋佳璇 , 王淑京 , 任志鸿 , 杜华茂 . 猪源益生芽孢杆菌的分离鉴定与生物学特性分析[J]. 动物营养学报, 2016 , 28(11) : 3542 -3548 . DOI: 10.3969/j.issn.1006-267x.2016.11.022

Abstract

The aim of this project was to isolate Bacillus spp with good performance from the normal intestinal flora of healthy swine, and to improve the feed utilization rate, reduce the infection of pig digestive tract and reduce the use of antibiotics. The feces specimen were collected from local back-yard cows which never feed with manufactured feeds, and the Bacillus spp with acid and bile salt tolerance was isolated from fresh feces. The isolates were then tested for the production potency of amylase, cellulose and protease on specific agar plates. The strains were identified by biochemical assays and 16s rDNA sequence alignment analysis. The results showed that 7 Bacillus strains were isolated which could secrete amylase, cellulose and proteinase. The survival ratios of Bacillus strains in pH 3.0 for 2 hours were 30% to 90%, and in 0.5% bile salt for 4 hours were 40% to 100%. The strains were identified as Bacillus stearothermophiluc (1 strain), Bacillus licheniformis (1 strain) and Bacillus subtilis (5 strains) by biochemical identification and molecular identification, respectively. In conclusion, it selects 7 strains identified Bacillus spp, which have good tolerance in low pH and high bile salt conditions, and the strains can secrete amylase, cellulose and proteinase.

Key words： probiotics; Bacillus spp; biochemical identification; 16S rDNA; enzyme production capacity

参考文献

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[13] 宁豫昌,赵绪永,郑鸣.猪源芽孢杆菌的分离鉴定及生物学特性研究[J].中国畜牧兽医,2012,39(9):65-68.

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