广西猪源益生菌的分离鉴定与生物学特性研究

doi:10.16431/j.cnki.1671-7236.2020.06.032

摘要/Abstract

摘要： 为了解广西地区猪粪便与微生态发酵饲料中益生菌的存在情况及合理开发和利用猪源益生菌，本试验共采集了15份猪粪便样品和5份益生菌发酵饲料并对其进行益生菌的分离，研究益生菌的形态学特性、生理生化特性及16S rRNA分子序列，进行生长曲线、耐胆盐试验、温度敏感试验、模拟胃肠道耐受试验、耐药性试验、体外抑菌试验、小鼠安全性试验、体内/外抑菌试验，对分离菌株进行生物学特性的研究分析。结果显示，本试验共分离鉴定出6种、29株益生菌，分别为1株屎肠球菌（C2）、2株乳酸肠球菌（C1、C3）、3株植物乳杆菌（R20、R30、R37）、4株干酪乳杆菌（R41~R44）、7株枯草芽孢杆菌（K1~K7）和12株蜡样芽孢杆菌（Y1~Y12）。从中筛选出6株具有生长性能好、耐胆盐、耐高温、对胃肠道耐受、体外抑菌能力强的益生菌：乳酸肠球菌C1、屎肠球菌C2、植物乳杆菌R20、干酪乳杆菌R41、枯草芽孢杆菌K1和蜡样芽孢杆菌Y3。将筛选出的具有优良特性的菌株进行小鼠安全性试验，结果表明，在10⁹ CFU/mL浓度下灌胃6株益生菌对小鼠是安全无毒害的。与此同时，C2、R20、R41、Y3能极显著提高小鼠日增重（P<0.01），K1能显著提高小鼠日增重（P<0.05）。体内抑菌试验结果表明，C2、R20、K1能降低沙门氏菌G21感染的小鼠的死亡率，有一定程度的体内抑菌作用。各项试验结果表明，R20和K1可作为猪源益生菌的选择菌株。

关键词: 益生菌; 分离; 生物学特性; 抑菌; 猪

Abstract: In order to investigate the prevalence of probiotics in pig feces and microecological fermented feeds in Guangxi area,and to rationally develop and utilize porcine probiotics,in this experiment,15 pig feces samples and 5 probiotic fermentation feeds were collected and the probiotic bacteria were isolated.The morphological characteristics,physiological and biochemical characteristics and 16S rRNA molecular sequence of the probiotics were studied.The growth curve,bile salt resistance test,temperature sensitivity test,gastrointestinal tolerance test,in vitro and in vivo bacteriostasis test,drug resistance test,mouse safety test were carried out to study the biological characteristics of the isolated strains.In this experiment,6 species and 29 probiotics strains were isolated and identified,including 1 Enterococcus faecium (C2),2 Enterococcus lactate (C1 and C3),3 Lactobacillus plantarum (R20,R30 and R37),4 Lactobacillus casei (R41-R44),7 Bacillus subtilis (K1-K7) and 12 Bacillus cereus (Y1-Y12).Six probiotics with good growth performance,bile salt resistance,high temperature resistance,gastrointestinal tolerance and strong antibacterial ability in vitro were screened out,including Enterococcus lactis C1,Enterococcus faecium C2,Lactobacillus plantarum R20,Lactobacillus casei R41,Bacillus subtilis K1 and Bacillus cereus Y3.The safety test of the selected strains in mice showed that six probiotics were safe and harmless to mice at the concentration of 10⁹ CFU/mL.At the same time,C2,R20,R41 and Y3 probiotics could extremely significantly increase the weight of mice (P<0.01),K1 could markedly increase the weight of mice (P<0.05).The results of bacteriostatic experiment in vivo showed that C2,R20 and K1 could reduce the mortality of mice infected by Salmonella G21,and had a certain degree of antibacterial effect in vivo.All test results showed that R20 and K1 could be selected as pig probiotic strains.

Key words: probiotics; isolation; biological characteristics; bacteriostasis; swine

中图分类号:

S816.3

张在, 郑瑞程, 方庆励, 黄伟坚. 广西猪源益生菌的分离鉴定与生物学特性研究[J]. 中国畜牧兽医, 2020, 47(6): 1921-1933.

ZHANG Zai, ZHENG Ruicheng, FANG Qingli, HUANG Weijian. Isolation,Identification and Biological Characteristics of Probiotics from Swine Origin in Guangxi[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(6): 1921-1933.

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