采用传统抑菌方法和体外生物膜模型,研究并筛选对大肠杆菌生物膜具有较强抑制能力的乳酸菌。结果表明,乳酸菌的抑菌性与抑制大肠杆菌生物膜形成能力之间并没有相关性。筛选得到了5株对保护人体免受大肠杆菌感染具有潜在应用价值的乳酸菌,即7-1、11-3、10-1、13-1、A-1,对生物膜形成量的抑制率分别为84.59%、 85.19%、70.09%、64.59%和65.20%,通过荧光显微镜观察发现它们均可以完全破坏生物膜的典型结构。通过耐酸耐胆盐实验,进一步筛选出了能很好适应胃肠环境压力的菌株,即菌株A-1(Lactobacillus acidophilus),为今后预防由大肠杆菌引起的感染提供了理论依据。
Traditional antibacterial method and in vitro biofilm model were used to select lactic acid bacteria (LAB) with strong inhibitory effects on Escherichia coli biofilm forming. The results showed that there was no correlation between LAB antibacterial activity and inhibition of E. coli biofilm formation. Five LAB isolates (7-1, 11-3, 10-1, 13-1, and A-1) that could potentially be used to protect humans against infections caused by E. coli showed significant inhibitory effects on the biofilm formation, with the inhibitory rates of 84.6%, 85.2%, 70.1%, 64.6% and 65.2%, respectively. The fluorescence microscope showed that all of these five isolates could destroy the biofilm structure completely. Of which, isolate A-1 (Lactobacillus acidophilus) was also acidic and bile salt tolerance. This study provides references for preventing E. coli infection in the future.
[1] COSTERTON J W, LEWANDOWSKI Z, DEBEER D, et al. Biofilms,the customized microniche [J].Journal of Bacteriology,1994,176(8): 2 137-2 142.
[2] KRZYSCIAK W, JURCZAK A,KOSCIELNIAK D, et al. The virulence of Streptococcus mutans and the ability to form biofilms[J].European Journal of Clinical Microbiology & Infectious Diseases,2014,33(4): 499-515.
[3] FUQUA W C,WINANS S C,GREENB E P. Quorum sensing in bacteria: The Luxl family of cell density-responsive transcriptional regulator[J].Joumal of Bacteriology,1994,176:269-275.
[4] WHITEHEAD N A, BARNARD A M L, SLATER H, et al. Quorum sensing in Gram-negative bacteria[J].FEMS Microbiology Reviews,2001,25(4):365-404.
[5] COSTERON J W, STEWART P S, GREENBERG E P. Bacterial biofilms:A common cause of persistent infections[J].Science,1999,284(5 418):1 318-1 322.
[6] PARSE M R, SINGH P K. Bacterial biofilms:an emerging link to disease pathogenesis[J]. Annual Review of Microbiology, 2003, 57(1):677-701.
[7] KOEBNICK C,WAGNER I, LEITZMANN P, et al. Probiotic beverage containing Lactobacillus casei Shirota improves gastrointestinal symptoms in patients with chronic constipation [J]. Canadian Journal of Gastroenterology,2003,17(11): 655-669.
[8] 宋雅珊,赵赫,李奕铮,等.米诺环素对大肠杆菌生物膜抑制作用研究[J].内科,2018,13(3):280-283.
[9] 陈盛,余加林,何念海,等.巯乙磺酸钠对大肠杆菌生物膜早期黏附及胞外聚合物的影响[J].第三军医大学学报,2011,33(24):2 554-2 557.
[10] 李晏乐,岳肖华,孙卫东,等.中草药抑菌作用的现代研究进展[J].中国中医基础医学杂志, 2018,24(6):860-862;874.
[11] 李斌,董明盛.黑木耳提取物对大肠杆菌生物膜形成的抑制作用[J].南京农业大学报,2010,23(13):36-37.
[12] 喻华英,何靖,等.五味中草药对大肠杆菌生物膜形成的影响[J].畜牧与兽医,2013,45(7):74-77.
[13] 刘叶,杨悦.我国抗生素滥用现状分析及建议[J].中国现代医生,2016,54(29):160-164.
[14] 李贺.芍药总多糖对口腔致龋细菌毒力因子作用的体外实验研究[D].乌鲁木齐:新疆医科大学, 2013.
[15] FIGUEROA-GONZALEZ I, QUIJANO G, RAMIREZ G, et al. Probiotics and prebiotics - perspectives and challenges[J]. Journal of the Science of Food and Agriculture,2011,91(8):1 341-1 348.
[16] 曹振辉,刘永仕,潘洪彬,等.乳酸菌的益生功能及作用机制研究进展[J].食品工业科技,2015,36(24):366-370;377.
[17] KHAN R, ZAKIR M, KHANAM Z, et al. Novel compound from Trachyspermum ammi (Ajowan caraway) seeds with antibiofilm and antiadherence activities against Streptococcus mutans: A potential chemotherapeutic agent against dental caries[J]. Journal of Applied Microbiology,2010,109(6):2 151-2 159.
[18] KANG M S, CHUNG J, KIM S M,et al. Effect of Weisselia cibaria isolates on the formation of Streptococcus mutans biofilm[J].Caries Research,2006,40(5):418-425.
[19] KOEBNICK C,WAGNER Ⅰ,LEITZMANN P, et al. Probiotic beverage containing Lactobacillus casei Shirota improves gastrointestinal symptoms in patients with chronic constipation[J].Canadian Journal of Gastroenterology,2003,17(11):655-669.
[20] 翟磊,凌空,宋振,等.哈萨克传统发酵食品中乳酸菌的分离鉴定及代谢特性研究[J].食品与发酵工业,2017, 43(7):122-127.
[21] 杨颖,陈卫,田丰伟,等. 产抑菌物质乳杆菌的筛选及性质的研究[J].工业微生物, 2006, 36(3): 13-17.
[22] O'TOOLE G A, PRATT L A, WATNICK P I, et al. Genetic approaches to study of biofilms[J]. Methods in Enzymology, 1999, 310(1):91-109.
[23] 姚沛琳.乳酸菌抑制变异链球菌生物膜形成的研究[D].无锡:江南大学, 2015.
[24] 李利,孔丽,张宁,等.耐酸耐胆盐乳酸菌的鉴定及筛选[J].食品科学,2015,36(21):123-128.
[25] 赵小茜,魏旭丹,陈戴玲,等.产多糖植物乳杆菌的耐酸耐胆盐能力[J].乳业科学与技术,2016,39(3):1-3.
[26] KHAN R,ZAKIR M,KHANAM Z,et al. Novel compound from Trachyspermum ammi (Ajowan caraway) seeds with antibiofilm and antiadherence activities against Streptococcus mutans: A potential chemotherapeutic agent against dental caries[J].Journal of Applied Microbiology,2010,109(6):2 151-2 159.
