甘草提取物对1株致犊牛腹泻大肠杆菌生物膜抑制作用的试验
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摘要
大肠杆菌生物膜的形成与其致病性、抗药性有密切关系,而中药在抗生物膜方面具有独特的优势。本试验对来自新疆阿克苏地区某奶牛场6份犊牛腹泻粪便进行了细菌分离鉴定,获得24株大肠杆菌,5株沙门菌。经96孔微量板法对已分离的24株大肠杆菌进行生物膜阳性菌筛选试验,获得生物膜阳性菌3株,其中1株为强阳性(XN-3-6株)。采用不同浓度甘草提取物对XN-3-6分离株生物膜形成抑制试验,结果表明:甘草提取物抑制大肠杆菌生物膜形成的最小浓度为5%。该试验结果为临床上防治大肠杆菌引起犊牛腹泻病提供了理论依据。
The Escherichia coli form biofilms, which are related to its pathogenicity, and drug resistance. Traditional Chinese medicine has unique advantages in anti-biofilm. We collected faeces from calf diarrhea in cow farm from Xinjiang Akshu. And then we screened bacteria by using conventional isolation and identification methods. TWe got 24 strains E. coli and 5 strains salmonella. Using 96 hole micro plate method, we found that of the 24 strains E. Coli, 3 strains formed biofilms, and one was strong positive(XN-3-6). We then tested if licorice extract could inhibit the formation of biofilm in E.coli(XN-3-6) by using different concentrations of licorice extract. The results showed that the licorice extract inhibited E.coli(XN-3-6) biofilm formation with a minimum concentration of 5%.
The Escherichia coli form biofilms, which are related to its pathogenicity, and drug resistance. Traditional Chinese medicine has unique advantages in anti-biofilm. We collected faeces from calf diarrhea in cow farm from Xinjiang Akshu. And then we screened bacteria by using conventional isolation and identification methods. TWe got 24 strains E. coli and 5 strains salmonella. Using 96 hole micro plate method, we found that of the 24 strains E. Coli, 3 strains formed biofilms, and one was strong positive(XN-3-6). We then tested if licorice extract could inhibit the formation of biofilm in E.coli(XN-3-6) by using different concentrations of licorice extract. The results showed that the licorice extract inhibited E.coli(XN-3-6) biofilm formation with a minimum concentration of 5%.
引文
[1] 刘栓奎,李明,党荣理.致病性大肠杆菌和出血性大肠杆菌研究进展[J].现代预防医学,2011,38(24):5 123-5 124.
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[4] O’Toole G A,Kolter R.Flagellar and twitching motility are necessary for Pseudomonas aeruginosa biofilm development[J].MolM icrobio,1988,30:295-304.
[5] OT’oole G,Kaplan H B,Kolter R.Biofilm formation asmicrobial development[J].Annu Rev M icrobio,2000,54:49-79.
[6] 孙晓梅,许杨.抗真菌中药的作用机理研究进展[J].中国中药杂志,2004,29(3):20-21.
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[17] 刘玉庆,李晔,车程川,等.大肠杆菌对中草药敏感性试验及其方法研究[J].中国兽医科技,2003,13(1) :23-24.
[18] 夏美玲,吕丽艳,刘野,等.5种中草药体外抑菌实验的研究[J].齐齐哈尔医学院学报,2009,46(24) :1 234-1 236.
[19] 官妍,程惠娟,汪长中,等.复方百肤青抗大肠杆菌引起细菌性阴道病生物膜的研究[J].中医药学刊,2004,22(9):82.
[20] 程惠娟,汪长中,官妍,等.细菌生物膜与细菌性阴道病复发相关性研究[J].热带病与寄生虫学报,2005(1):5-7.
[21] 王艳,汪本凡,程惠娟.复方百肤青抗细菌性阴道病生物膜的研究[J].中医药学刊,2006,24(3):3-5.
[22] 官妍,程惠娟,汪长中,等.百肤青对细菌性阴道病主要致病菌抗菌作用的研究[J].中医药学刊,2004,22(10):1 835.
[23] 官妍,程惠娟,汪长中,等.中药复方百肤青对细菌性阴道病生物膜渗透性的影响[J].辽宁中医杂志,2007,34(2):236-237.
[24] 丁长玲,邱世翠,宫照龙,等.甘草的体外抑菌作用研究[J].时珍国医国药,2002,13(9):518.
[25] 吴传茂,吴周和,陈雄.甘草提取液的抑菌作用研究[J].中国商办工业,2000(4):49-50.
[26] Cui Y M,Yu L J,Ao M Z,et al.Studies on flavonoids extraction technology from Glycyrrhiza inflata and their bacteriostatic activities[J].Zhong Yao Cai,2006,29(8):38-41.
[27] Li W,Asada Y,Yoshikawa T.Antimicrobial flavonoids from Glycyrrhiza glabra hairy root cultures[J].Planta Med,1998,64(8):6-7.
[28] 喻华英,贾桂珍,邹开胜.犊牛腹泻病中大肠杆菌的分离与鉴定[J].黑龙江畜牧兽医,2006(6):67-68.
[29] 孙伟,刘晓梅,周峰,汤仁仙.大肠杆菌致小鼠急性腹膜炎模型的建立[J].齐齐哈尔医学院学报,2008,17:2 049-2 051.
[2] 陈世德,时常仁.致病性大肠杆菌的研究进展与临床[J].白求恩医科大学学报,1981,7(4):118-123.
[3] Cvikovitch D G,L iY H,Ellen R P.Quonm sensing and biofim for mation in streptococcal infections[J].JCl in Invest,2003,112(11):1 626-1 632.
[4] O’Toole G A,Kolter R.Flagellar and twitching motility are necessary for Pseudomonas aeruginosa biofilm development[J].MolM icrobio,1988,30:295-304.
[5] OT’oole G,Kaplan H B,Kolter R.Biofilm formation asmicrobial development[J].Annu Rev M icrobio,2000,54:49-79.
[6] 孙晓梅,许杨.抗真菌中药的作用机理研究进展[J].中国中药杂志,2004,29(3):20-21.
[7] 段高飞,韩峰,李京宝,等.细菌生物膜相关感染的防治方法研究进展[J].中国海洋大学学报,2010,40(5) :107-111.
[8] RobVan H,Chris W.Michiels.Role of bacterial cell surface structures in Escherichia coli biofilm formation[J].Research in M icrobiology,2005,156:626-633.
[9] Leslie A P,Roberto K.Genetic analysis of Escherichia coli biofilm formation:roles of flagella.Motility chemotaxis and type I pili[J].M o lM icrob io,1998,30:285-293.
[10] Fernandez L A,Berenguer J.Secretion and assembly of regular surface structures in Gram-negative bacteria[J].FEM S M icrob iol Rev,2000,24:21-44.
[11] Soutourina O A,Bertin P N.Regulation cascade of flagellar expression in Gram-negative bacteria[J].FEMS M icrobiol Rev,2003,27:505-523.
[12] 苏建青,褚秀玲,刘文强,等.中草药防治鸡大肠杆菌病的机理和临床应用效果[J].湖北农业科学,2009,48(8) :124-125.
[13] 刘巧,骆伟键.注射用双黄连对细菌生物膜影响研究[J].中国民族民间医药,2010,(6):12-13.
[14] 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].南京农业大学学报,2010,23(13):36-37.
[15] 李淑梅,杨帆.中草药提取物对大肠杆菌的体外抑菌试验[J].安徽农业科学,2006,34(23) :6 212-6 222.
[16] 卫文强,吴润,刘磊.四味中草药体外抗菌作用研究[J].安徽农业科学,2009,37(6) :2 530-2 531.
[17] 刘玉庆,李晔,车程川,等.大肠杆菌对中草药敏感性试验及其方法研究[J].中国兽医科技,2003,13(1) :23-24.
[18] 夏美玲,吕丽艳,刘野,等.5种中草药体外抑菌实验的研究[J].齐齐哈尔医学院学报,2009,46(24) :1 234-1 236.
[19] 官妍,程惠娟,汪长中,等.复方百肤青抗大肠杆菌引起细菌性阴道病生物膜的研究[J].中医药学刊,2004,22(9):82.
[20] 程惠娟,汪长中,官妍,等.细菌生物膜与细菌性阴道病复发相关性研究[J].热带病与寄生虫学报,2005(1):5-7.
[21] 王艳,汪本凡,程惠娟.复方百肤青抗细菌性阴道病生物膜的研究[J].中医药学刊,2006,24(3):3-5.
[22] 官妍,程惠娟,汪长中,等.百肤青对细菌性阴道病主要致病菌抗菌作用的研究[J].中医药学刊,2004,22(10):1 835.
[23] 官妍,程惠娟,汪长中,等.中药复方百肤青对细菌性阴道病生物膜渗透性的影响[J].辽宁中医杂志,2007,34(2):236-237.
[24] 丁长玲,邱世翠,宫照龙,等.甘草的体外抑菌作用研究[J].时珍国医国药,2002,13(9):518.
[25] 吴传茂,吴周和,陈雄.甘草提取液的抑菌作用研究[J].中国商办工业,2000(4):49-50.
[26] Cui Y M,Yu L J,Ao M Z,et al.Studies on flavonoids extraction technology from Glycyrrhiza inflata and their bacteriostatic activities[J].Zhong Yao Cai,2006,29(8):38-41.
[27] Li W,Asada Y,Yoshikawa T.Antimicrobial flavonoids from Glycyrrhiza glabra hairy root cultures[J].Planta Med,1998,64(8):6-7.
[28] 喻华英,贾桂珍,邹开胜.犊牛腹泻病中大肠杆菌的分离与鉴定[J].黑龙江畜牧兽医,2006(6):67-68.
[29] 孙伟,刘晓梅,周峰,汤仁仙.大肠杆菌致小鼠急性腹膜炎模型的建立[J].齐齐哈尔医学院学报,2008,17:2 049-2 051.