牛源产酸芽孢杆菌的分离筛选及益生特性研究  

Isolation and Screening of Acid-producing Bacillus from Cattle and Study on Its Prebiotics

作　　者：王婷 安明轩 李正浩 胡瑞瑞 王悦[1] 倪伟[1] 胡圣伟[1] WANG Ting;AN Ming-xuan;LI Zheng-hao;HU Rui-rui;WANG Yue;NI Wei;HU Sheng-wei(College of Life Science,Shihezi University,Shihezi,Xinjiang 832000,China)

机构地区：[1]石河子大学生命科学学院,新疆石河子832000

出　　处：《动物医学进展》2023年第10期12-18,共7页Progress In Veterinary Medicine

基　　金：兵团“强青”科技创新人才项目(2021CB033);兵团英才第二周期第一层次培养计划。

摘　　要：为了获得牛源具有益生特性良好、抗病原菌且安全的芽孢杆菌优势菌株,以在实际生产应用中发挥其替抗作用。通过采集健康牛肠道内容物,经过培养基初筛和纯化,16S rRNA基因扩增测序对分离菌株进行分子学鉴定,对鉴定得到的菌株进行耐酸、耐胆盐、耐人工肠胃液、自聚集率、产水解酶、致病菌抑制能力及抗生素敏感度测定。共分离出21株菌株,为了评价其益生特性,筛选出了3株益生菌为地衣芽孢杆菌BL2、解淀粉芽孢杆菌BA11和贝莱斯芽孢杆菌BV17,在强酸高胆盐和人工模拟肠胃液具有较高耐受性,自聚集率最低为57.90%,能产生淀粉酶和纤维素酶;地衣芽孢杆菌BL2只对金黄色葡萄球菌有抑制作用,解淀粉芽孢杆菌BA11和贝莱斯芽孢杆菌BV17对金黄色葡萄球菌、沙门氏菌、大肠埃希氏菌均有抑制作用;对常见抗菌药物敏感,不产生耐药性。在小鼠试验中,混合灌胃28 d,小鼠未出现不良反应,精神正常,无腹泻情况和中毒症状并且小鼠体重有所增加。表明3株芽孢杆菌为安全且具有增重作用的益生菌,其作为潜在益生菌可为牛源益生菌制剂的研究提供候选菌株。The aim of the trial was to obtain superior strains of bovine origin with good probiotic properties,resistant to pathogenic bacteria and safe for Bacillus,in order to use them as alternative resistance in practical applications.In this study,intestinal contents of healthy cattle were collected,initially screened and purified in the media,and 16S rRNA gene amplification sequencing was performed for molecular identification of the isolated strains.Acid and bile salt resistance,artificial intestinal fluid resistance,self-aggregation rate,hydrolytic enzyme production,pathogen inhibition ability and antimicrobial sensitivity were measured for the identified strains.A total of 21 strains were isolated.In order to evaluate their probiotic properties,3 strains were screened out as Bacillus licheniformis BL2,Bacillus amyloliquefaciens BA11 and Bacillus velezensis BV17.They had high tolerance in strong acid and high bile salt and artificial intestinal fluid,the lowest self-aggregation rate was 57.90%.All three Bacillus strains can produce starch hydrolase and cellulose hydrolase;Bacillus licheniformis BL2 only had inhibitory effects on Staphylococcus aureus,while Bacillus amyloliquefaciens BA11 and Bacillus velezensis BV17 had inhibitory effects on Staphylococcus aureus,Salmonella and Escherichia coli.It is sensitive to common antibiotics and does not develop resistance.In the mouse experiment,the mice showed no adverse reactions and were mentally normal for 28 days after mixed gavage.There were no diarrhea and poisoning symptoms,and the mice gained weight.The three strains of Bacillus were safe and had the effect of weight gain.As potential probiotics,they could provide candidate strains for the study of bovine probiotics.

关 键 词：肠道微生物 益生菌 芽孢杆菌 分离筛选 抗生素药敏试验 

分 类 号：S852.616[农业科学—基础兽医学] S858.23[农业科学—兽医学]