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空肠弯曲菌在光滑表面形成生物膜的模式。

The mode of biofilm formation on smooth surfaces by Campylobacter jejuni.

作者信息

Moe Kyaw Kyaw, Mimura Junichiro, Ohnishi Takahiro, Wake Tomoya, Yamazaki Wataru, Nakai Masaaki, Misawa Naoaki

机构信息

Laboratory of Veterinary Public Health, Department of Veterinary Science, Faculty of Agriculture, University of Miyazaki.

出版信息

J Vet Med Sci. 2010 Apr;72(4):411-6. doi: 10.1292/jvms.09-0339. Epub 2009 Dec 10.

Abstract

Many microorganisms produce extracellular polymers referred to collectively as "slime" or glycocalyx, and form biofilms on solid surfaces in natural ecosystems. Campylobacter jejuni, one of the most important foodborne pathogens, also has the ability to form biofilm on stainless steel, glass, or polyvinyl chloride in vitro. However, the issue of biofilm formation by Campylobacter species has not been extensively examined. The present study was performed to examine the mode of adhesion of C. jejuni to a smooth surface. When bacterial suspensions in Brucella broth were incubated in microplate wells with a glass coverslip, microcolonies 0.5~2 mm in diameter were formed on the coverslip within 2 hr from the start of incubation. These microcolonies gradually grew and formed a biofilm of net-like connections within 6 hr. Transmission electron microscopy indicated that massive amounts of extracellular material masked the cell surface, and this material bound ruthenium red, suggesting the presence of a polysaccharide moiety. Scanning electron microscopy indicated that the flagella acted as bridges, forming net-like connections between the organisms. To determine the genes associated with biofilm formation, aflagellate (flaA(-)) and flagellate but non-motile (motA(-)) mutants were constructed from strain 81-176 by natural transformation-mediated allelic exchange. The flaA(-) and motA(-) mutants did not form the biofilm exhibited by the wild-type strain. These findings suggest that flagella-mediated motility as well as flagella is required for biofilm formation in vitro.

摘要

许多微生物会产生统称为“黏液”或糖萼的细胞外聚合物,并在自然生态系统中的固体表面形成生物膜。空肠弯曲菌是最重要的食源性病原体之一,在体外也有在不锈钢、玻璃或聚氯乙烯表面形成生物膜的能力。然而,弯曲菌属生物膜形成的问题尚未得到广泛研究。本研究旨在检测空肠弯曲菌在光滑表面的黏附方式。当将布鲁氏菌肉汤中的细菌悬液与盖玻片一起在微孔板孔中孵育时,从孵育开始2小时内,盖玻片上就形成了直径为0.5至2毫米的微菌落。这些微菌落逐渐生长,并在6小时内形成了网状连接的生物膜。透射电子显微镜显示,大量细胞外物质覆盖了细胞表面,且这种物质与钌红结合,表明存在多糖部分。扫描电子显微镜显示,鞭毛起到桥梁作用,在菌体之间形成网状连接。为了确定与生物膜形成相关的基因,通过自然转化介导的等位基因交换,从81-176菌株构建了无鞭毛(flaA(-))和有鞭毛但无运动能力(motA(-))的突变体。flaA(-)和motA(-)突变体均未形成野生型菌株所呈现的生物膜。这些发现表明,鞭毛介导的运动能力以及鞭毛对于体外生物膜的形成是必需的。

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