鸡源高黏附乳酸菌的分离筛选及主要生物学特性研究  

Isolation,Screening and Characteristic Analysis on High Adhesion Lactic Acid Bacteria from Chicken

作　　者：范雪莲 张知之 冯茜茜 严丹丽 薛念宇 刘丹丹[1,2] 候照峰 许金俊 陶建平[1,2] FAN Xueian;ZHANG Zhizhi;FENG Qianqian;YAN Dani;XUE Nianyu;LIU Dandan;HOU Zhaofeng;XU Jinjun;TAO Jianping(College of Veterinary Medicine,Yangzhou University,Yangzhou,Jiangsu 225009;Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses,Yangzhou,Jiangsu 225009)

机构地区：[1]扬州大学兽医学院,江苏扬州225009 [2]江苏高校动物重要疫病与人兽共患病防控协同创新中心,江苏扬州225009

出　　处：《中国家禽》2023年第12期67-74,共8页China Poultry

基　　金：国家自然科学基金项目(31972698);江苏省高校优势学科建设三期工程项目;高等学校学科创新引智计划(D18007)。

摘　　要：研究旨在筛选高黏附特性的鸡源乳酸菌,评价其主要生物学特性。试验应用MRS琼脂培养基从SPF鸡十二指肠、盲肠及小肠分离菌株,通过形态学观察、生理生化试验进行初步筛选,随后采用ERIC-PCR与16S rRNA基因序列分析相结合的方法对分离菌株进行种属鉴定,并对分离鉴定的乳酸菌株进行黏附能力、耐酸与耐胆汁特性、生长曲线、体外传代能力等生物学特征进行分析。结果显示:在34株分离菌株中鉴定到9种基因型,其中7株为罗伊氏乳杆菌、2株为约氏乳杆菌。菌株编号为C7的罗伊氏乳酸杆菌对HT-29细胞黏附率为(32.3±1.67)%,显著高于商品化乳酸乳球菌MG1363株和乳杆菌BNCC182567株以及除S3株外的其他分离株(P<0.05);在pH2.0~5.0条件下存活率均高于100%,在40%与60%新鲜鸡胆汁作用下存活率分别为(116.67±11.89)%和(102.33±10.87)%,在体外连续传代10代仍能保持良好的活力。结果表明分离获得的C7分离株为罗伊氏乳酸杆菌,具有高黏附特性、耐酸与耐胆汁特性,该菌株为研究益生菌制剂及口服疫苗表达载体奠定了初步基础。This study was to screen lactic acid bacteria with high adhesion properties from chicken intestine and evaluate its main biological characteristics.Bacterial strains were isolated from duodenal,cecum and small intestine of SPF chickens using MRS agar medium,and were preliminary screened based on the results of morphological observation,physiological and biochemi-cal tests.Subsequently,ERIC-PCR was innovatively combined with 16S rRNA gene sequence analysis method,applied to the species identification of natural lactic acid bacteria.The biological characteristics of the isolated strains,such as adhesion and colonization ability,acid and bile tolerance,growth curve and continuous passage in vitro,were analyzed.The results showed that 9 genotypes were identified among the 34 isolated strains,of which seven strains were Lactobacillus reuteri and two strains were L.Johnsonii.The adhesion rate of L.reuteri No.C7 strain to HT-29 cells was significantly higher than that of commercial Lactococ-cus lactis MG1363 strain,Lactobacillus BNCC182567 strain and other isolates except for S3 strain(P<0.05);Its survival rate was higher than 100%under the condition of pH 2.0~5.0,and(116.67±11.89)%and(102.33±10.87)%when treated with 40%and 60%fresh chicken bile,respectively.C7 strain could maintain good vitality after continuous passage for 10 generations in vitro.The results revealed that C7 strain was Lactobacillus reuteri,had the highest adhesion to HT-29 cells,acid and bile tolerance,and maintained good growth performance after continuous passage in vitro.This strain could be used for the study of probiotic preparations and oral vaccine expression vectors such as coccidiosis.

关 键 词：鸡源 乳酸菌 分离 鉴定 黏附性 

分 类 号：S852.6[农业科学—基础兽医学]