牛源乳酸菌的分离鉴定及体外益生特性研究  

Isolation and Identification of Lactic Acid Bacteria from Calves andTheir Probiotic Properties in Vitro

作　　者：李元元 李鑫[2] 何泽纯 张文举[1] LI Yuanyuan;LI Xin;HE Zechun;ZHANG Wenju(College of Animal Science and Technology,Shihezi University,Shihezi 832000,China;College of Life Sciences,Shihezi University,Shihezi 832000,China)

机构地区：[1]石河子大学动物科技学院,石河子832000 [2]石河子大学生命科学学院,石河子832000

出　　处：《动物营养学报》2024年第1期626-636,共11页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：新疆生产建设兵团重大科技项目(2021AA004)。

摘　　要：本研究旨在筛选益生特性优良的牛源罗伊氏乳杆菌和约氏乳杆菌,为后期微生态制剂的开发和利用奠定理论依据。试验以粪菌移植后的断奶中国荷斯坦犊牛直肠粪便为菌株分离来源,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 rDNA测序分离鉴定得到14株罗伊氏乳杆菌和1株约氏乳杆菌。通过产酸能力、耐酸和耐胆盐性能、疏水性能、抑菌性能及抗生素敏感性试验,评估其益生特性。结果表明:各菌株具有较好的产酸能力,培养24 h后,菌液pH集中在4.0~4.5;在pH为4.0时,各菌株的存活率均超过80%;在0.3%的胆盐环境中,各菌株存活率均在75%以上,其中L81和L29的菌株存活率较高,分别为88.40%和86.87%;在疏水性能方面,L29、L39、L65、L74、L81、L102、L107、L116和L120疏水率均在55%以上,具有良好的疏水性能;15株菌株均表现出一定的抑菌作用,其中L10和L81对金黄色葡萄球菌、大肠杆菌及沙门氏菌整体抑菌效果较好;L29、L35和L81对5种抗生素中的至少3种表现出的敏感性要高于其他菌株。综上所述,本试验筛选出的14株罗伊氏乳杆菌和1株约氏乳杆菌具有较好的益生特性,尤其是罗伊氏乳杆菌L81和约氏乳杆菌L29,可作为后续开发为牛源乳酸菌微生态制剂的备用菌株。The purpose of this study was to screen Lactobacillus reuteri and Lactobacillus johnsonii with excellent probiotic properties,and to lay a theoretical basis for the development and utilization of probiotics in the later stage.In this experiment,the rectal feces of weaned Chinese Holstein calves after fecal microbiota transplantation were used as the source of strain isolation.Lactic acid bacteria were isolated and purified by MRS solid medium containing calcium carbonate,and 14 strains of Lactobacillus reuteri and 1 strain of Lactobacillus johnsonii were isolated and identified by physiological and biochemical and 16S rDNA sequencing.The probiotic properties were evaluated by acid production,acid and bile salt resistance,hydrophobicity,antibacterial ability and antibiotic sensitivity tests.The results showed that each strain had good acid production ability.After 24 h of culture,the pH in the bacterial solution was concentrated between 4.0 and 4.5.When pH was 4,the survival rate of each strain was more than 80%.In 0.3%bile salt environment,the survival rate of each strain was more than 75%,and the survival rates of L81 and L29 were higher,which were 88.40%and 86.87%,respectively.In terms of hydrophobicity,the hydrophobic rates of L29,L39,L65,L74,L81,L102,L107,L116 and L120 were all above 55%,showing good hydrophobicity.All the 15 strains showed certain antibacterial effects,among which L10 and L81 had better antibacterial effects on Staphylococcus aureus,Escherichia coli and Salmonella.L29,L35 and L81 showed higher sensitivity to at least three of the five antibiotics than the other strains.In summary,the 14 strains of Lactobacillus reuteri and 1 strain of Lactobacillus johnsonii screened in this experiment have good probiotic properties,especially Lactobacillus reuteri L81 and Lactobacillus johnsonii L29,which can be used as backup strains for subsequent development of bovine lactic acid bacteria probiotics.

关 键 词：荷斯坦犊牛 乳酸菌 分离 鉴定 益生特性 

分 类 号：S811[农业科学—畜牧学]