三株乳酸杆菌的分离鉴定与益生特性研究  

Study on the isolation,identification of three strains of Lactobacilli and their probiotic properties

作　　者：梁东梅 李玉鹏[2] 杜艳芬 朱琪[2] 安建勇[2] 崔茂盛[2] 李治欣[2] 乔家运[2] 杨卓婧 王文杰[2] 李海花 LIANG Dongmei1,2,LI Yupeng2,DU Yanfen3,ZHU Qi2,AN Jianyong2,CUI Maosheng2,LI Zhixin2,QIAO Jiayun2,YANG Zhuojing4,WANG Wenjie2,L I Haihua12＊(l.Tianjin Normal University,Tianjin 300387,C hina;2.Tianjin Institute of Animal Husbandry and Veterinary Science,Tianjin 300381 ,C hina;3. Thermo Fisher Scientific （Shanghai） Instrument Co., LTD, Shanghai 201206, China,4. The old city of Luoyang Institute of Animal Health Supervision, Luoyang,Henan Province 471000,China)

机构地区：[1]天津师范大学生命科学学院,天津300387 [2]天津市畜牧兽医研究所,天津300381 [3]赛默飞世尔(上海)仪器有限公司,上海201206 [4]河南省洛阳市老城区动物卫生监督所,河南洛阳471000

出　　处：《中国饲料》2018年第11期24-29,共6页China Feed

基　　金：国家自然科学基金(31702147)

摘　　要：本研究通过分离鉴定猪肠道内乳酸杆菌并对其益生特性进行比较分析,旨在为乳酸杆菌在仔猪饲粮中的应用提供科学依据。试验选取健康猪粪便,利用选择性培养基进行厌氧培养和单克隆纯化,共获得三株乳酸杆菌,通过生化鉴定、16Sr RNA基因序列测定和同源性分析,确定分离菌株分别为植物乳杆菌、唾液乳杆菌、嗜酸乳杆菌;采用牛津杯法、活菌计数法和细胞试验等研究三株乳酸杆菌体外抑菌活性、黏附性及竞争性黏附猪小肠上皮细胞(IPEC-J2)的益生特性。结果表明:与对照组相比,三株乳酸杆菌培养上清液的抑菌圈直径(P<0.01)和抑菌效果(P<0.01)差异均极显著,并且以植物乳杆菌培养上清液的抗菌效果为最佳;三株乳酸杆菌均可黏附于IPEC-J2细胞,并且以植物乳杆菌黏附IPEC-J2的菌数为最多;与对照组相比,三株乳酸杆菌对大肠杆菌黏附IPEC-J2细胞均具有抑制作用(P<0.01),并且以植物乳杆菌抑制黏附的效果最好。In this study,we isolated and identified Lactobacilli from pig gut and compared their probiotic properties to provide a scientific basis for the application of lactic acid bacteria in piglet diets.Three strains of Lactobacilli were obtained from healthy pig feces by being purified under anaerobic environment.These three isolates were identified as L.plantarum,Lactobacillus salivarius and Lactobacillus acidophilus by biochemical test,16S rRNA gene sequencing and homology analysis.The probiotic properties of the three isolates in vitro,in-cluding antimicrobial activity,adhesion and competitive adherence to porcine intestinal epithelial cells （IPEC-J2）were studied by using Oxford cup method,live bacterial count method and cell assay.Compared with the control group,the antibacterial circle diameter （P〈0.01） and the bacteriostatic effect （P〈0.01） of the culture supernatants from the tree isolates showed significant difference,and the antibacterial activity of the culture su-pernatant from L.plantarum was the best.The three strains of Lactobacilli could adhere to IPEC-J2 cells,the number of L.plantarum adherence to IPEC-J2 cells was the most.Compared with the control group,the three strains of lactic acid bacteria have a significant inhibitory effect on the adhesion cells of Escherichia coli （P〈0.01）,and this inhibitory effect on the adhesion caused by L.plantarum was the best.

关 键 词：乳酸杆菌 分离鉴定 猪小肠上皮细胞 益生特性 

分 类 号：S816.3[农业科学—饲料科学]