大肠埃希菌生物被膜体外模型的建立及其应用  

Establishment and Application of Escherichia coli Biofilm Model in vitro

作　　者：陈朝喜[1,2] 朱恒乾[1] 廖晓萍[1] 刘雅红[1] 

机构地区：[1]华南农业大学兽医学院,广东广州510642 [2]西南民族大学生命科学与技术学院,四川成都610041

出　　处：《动物医学进展》2011年第3期8-12,共5页Progress In Veterinary Medicine

基　　金：国家自然科学基金-广东省联合基金项目(U0631006)

摘　　要：以ATCC25922为研究对象,采用改良结晶紫染色、快速银染法和扫描电镜技术对其生物被膜进行定量和定性研究,建立大肠埃希菌生物被膜体外模型,利用优化的模型条件对249株临床分离株进行验证。采用琼脂平板计数法绘制大肠埃希菌浮游菌和被膜菌生长曲线,比较其生长特性的异同。对不同成膜能力的大肠埃希菌临床分离株进行色氨酸定量试验,分析不同培养条件对生物被膜菌胞外基质分泌量的影响。快速银染法和扫描电镜定性研究结果表明,大肠埃希菌形成生物被膜后,形成一种浮游细菌和生物被膜细菌共同分布于同一生存空间内的特殊生理结构;不同稀释度的接种量对细菌生物被膜生物量的变化影响不显著(P>0.05)。249株大肠埃希菌分为强成膜能力、中等成膜能力、弱成膜能力和无成膜能力四个表型,分别占3.21%,18.88%,51.81%和26.10%。To establish the Escherichia coli biofilm model in vitro,the reference strain ATCC25922 was chosen for quantitative and qualitative analysis using modified crystal violet staining,rapid silver staining and scanning electron microscopy methods,and 249 clinical strains were tested for biofilm-forming ability analysis.Plate count method was used for growth curve mapping of Escherichia coli planktonic and biofilm bacteria.And tryptophan quantitative experiment was used to compare the impact of different culture conditions on extracellular matrix secretion of different biofilm-forming ability Escherichia coli.Rapid silver staining and scanning electron microscopy results showed that after biofilm formation,E.coli formed a special growth pattern which biofilm and planktonic bacteria co-located in the same living space.Different dilution of the bacterial inoculum on biofilm biomass change is not obvious（P0.05）.The results showed that 249 E.coli were divided into four groups with strong,moderate,weak or absent biofilm-forming ability,accounting for 3.21%,18.88%,51.81% and 26.10%,respectively.

关 键 词：改良结晶紫染色 快速银染法 扫描电镜 生物被膜 色氨酸定量试验 

分 类 号：S852.612[农业科学—基础兽医学]