动物营养学报 >

2021 , Vol. 33 >Issue 6: 3365 - 3379

DOI: https://doi.org/10.3969/j.issn.1006-267x.2021.06.038

反刍与草食动物营养与饲料 RUMINANT AND HERBIVORE NUTRITION AND FEED

羊瘤胃源乳酸菌的分离鉴定及其生物学特性分析

  • 何江波 ,
  • 姚志芳 ,
  • 吴国芳 ,
  • 杨雨鑫 ,
  • 王磊
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  • 1. 西北农林科技大学动物科技学院, 杨凌 712100;
    2. 青海大学畜牧兽医科学院, 西宁 810016
何江波(1994-),男,重庆人,硕士研究生,研究方向为饲料资源开发与利用。E-mail:hjb0625@126.com

收稿日期: 2020-12-02

  网络出版日期: 2021-06-10

基金资助

青海省重点研发与转化计划（2019-NK-173，2020-NK-127）；新疆生产建设兵团重点领域科技攻关计划（2020AB016）；国家绒毛用羊产业技术体系（CARS-39-12）

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Isolation and Identification of Lactic Acid Bacteria from Sheep Rumen and Analysis of Their Biological Characteristics

  • HE Jiangbo ,
  • YAO Zhifang ,
  • WU Guofang ,
  • YANG Yuxin ,
  • WANG Lei
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  • 1. College of Animal Science and Technology, Northwest A&F University, Yangling 712100, China;
    2. Academy of Animal Science and Veterinary Medicine, Qinghai University, Xining 810016, China

Received date: 2020-12-02

  Online published: 2021-06-10

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摘要

本研究旨在探寻益生特性优良、可作为抗生素替代品的乳酸菌，并为后续应用于畜禽养殖提供理论依据。本试验以青海黑藏羊和西藏白绒山羊瘤胃内容物为菌株分离来源，采用含有碳酸钙的MRS固体培养基分离纯化乳酸菌，经生理生化和16S rRNA测序鉴定及产酸产气试验分离得到22株同型发酵乳酸菌，测定并评价其生长速率、产酸能力、耐酸和耐胆盐性能、抑菌能力、自凝集率及抗生素敏感性。综合比较不同菌株之间的生物学特性，分别在片球菌属、肠球菌属、乳杆菌属中筛选获得2株戊糖片球菌、1株海氏肠球菌、2株戊糖乳杆菌。结果表明：2株戊糖片球菌均在4 h后进入对数生长期，16 h达到稳定期，24 h后菌液pH达到4.0以下，能耐受pH 3.0和0.3%的胆盐环境；1株海氏肠球菌在2 h后进入对数生长期，12 h达到稳定期，24 h后pH达到4.5以下，能耐受pH 4.0和0.3%的胆盐环境；2株戊糖乳杆菌均在2 h后进入对数生长期，16 h达到稳定期，16 h后pH达到4.0以下，能耐受pH 3.0和0.3%的胆盐环境，5株乳酸菌自凝集率、抑菌能力及安全性均表现优良。综上所述，本试验筛选获得的5株不同种属乳酸菌生物学特性良好，可作为后续开发微生物发酵饲料及动物微生态制剂的菌种来源。

关键词： 羊; 瘤胃; 乳酸菌; 分离; 鉴定; 生物学特性

本文引用格式

何江波 , 姚志芳 , 吴国芳 , 杨雨鑫 , 王磊 . 羊瘤胃源乳酸菌的分离鉴定及其生物学特性分析[J]. 动物营养学报, 2021 , 33(6) : 3365 -3379 . DOI: 10.3969/j.issn.1006-267x.2021.06.038

Abstract

This study aimed to quest several excellently probiotic lactobacilli which could substitute antibiotics, and provide theoretical support for subsequent application in livestock and poultry breeding. In this study, rumen contents collected from Qinghai black Tibetan sheep and Tibetan white cashmere goats were taken as source bacteria strains. The lactobacilli were isolated and purified using MRS solid medium with CaCO3. A total of 22 strains of homotypic fermenting lactobacilli were obtained after a series of physiological-biochemical tests, 16S rRNA sequencing identification, and acid and gas productivity tests. Determination and evaluation tests were proceeded with regards to growth rates, acidogenicity, tolerance to acid and bile salt, bacteriostatic performance, self-agglutination rate, as well sensitivity to antibiotics. Five bacterial strains were screened from the species of Pediococcus, Enterococcus, and Lactobacillus after a comparison on biological features of these strains. The screened strains consisted of two stains of Pediococcus pentosaceus, one strain of Enterococcus hirschii, two strains of Lactobacillus pentosus. The results showed that both strains of Pediococcus pentosaceus entered logarithmic growth phase after 4 h of cultivation, and reached stable phase after 16 h; the pH of bacteria solutions went below 4.0 after 24 h and it was observed that both strains could tolerate a pH of 3.0 and a bile salt environment of 0.03%. The only strain of Enterococcus hirschii entered the algorithmic growth phase after 2 h of cultivation, and reached stable phase after 12 h; the pH of bacteria solutions went below 4.5 after 24 h and it was observed that bacterial strain could tolerate a pH of 4.0 and a bile salt environment of 0.3%. The two strains of Lactobacillus pentosus entered the algorithmic growth phase after 2 h of cultivation, and reached stable phase after 16 h; the pH of bacteria solutions went below 4.0 after 16 h and it was observed that bacterial strain could tolerate a pH value of 3.0 and a bile salt environment of 0.3%. This study found that all of the five lactobacilli from different species had excellent self-agglutination rate, bacteriostatic performance, and safety. In conclusion, this 5 strains are believed promising source strains for the follow-up production of micro-organism fermented feeds and micro-ecological preparation products for animals considering their excellent biological characteristics.

Key words： sheep; rumen; lactic acid bacteria; isolation; identification; biological characteristics

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