拉曼光谱技术分析副溶血性弧菌生物被膜胞外聚合物的变化

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• 英文论文题名：Changes of extracellular polymer substances in the formation of Vibrio parahaemolyticus biofilm using raman spectroscopy analysis
• 论文作者：赵飞 ; 赵爱静 ; 张昭寰 ; 孙晓红 ; 潘迎捷 ; 赵勇
• 英文论文作者：Zhao Fei ; Zhao Aijing ; Zhang Zhaohuan ; Sun Xiaohong ; Pan Yingjie ; Zhao Yong ; College of Food Science and Technology ; Shanghai Ocean University ; Shanghai Engineering Research Center of Aquatic-Product Processing & Preservation ; Laboratory of Quality & Safety Risk Assessment for Aquatic Product on Storage and Preservation(Shanghai) ; Ministry of Agriculture
• 年：2016
• 作者机构：上海海洋大学食品学院;上海水产品加工及贮藏工程技术研究中心;农业部水产品贮藏保鲜质量安全风险评估实验室(上海);
• 论文关键词：拉曼光谱 ; 生物被膜 ; 胞外聚合物 ; 副溶血性弧菌
• 英文论文关键词：raman scattering ; biofilm ; EPS ; vibrio parahaemolyticus
• 会议召开时间：2016-11-09
• 会议录名称：中国食品科学技术学会第十三届年会论文摘要集
• 英文会议录名称：Abstracts of the 13th Annual Meeting of CIFST
• 语种：中文
• 分类号：R378;O657.37
• 学会代码：ZGSP
• 会议名称：中国食品科学技术学会第十三届年会
• 会议地点：中国北京
• 主办单位：中国食品科学技术学会
• 学会名称：中国食品科学技术学会
• 页数：2
• 文件大小：115k
• 原文格式：O
• 会议级别：全国

摘要

拉曼光谱技术已经被证明是一种高效实用的技术可用于分析生物被膜胞外聚合物的化学成份。本文以首要食源性致病菌副溶血性弧菌在玻璃材料表面形成的生物被膜为研究对象,用拉曼光谱技术分析了胞外聚合物的化学成分,并用激光共聚焦显微镜和扫描电镜观察了不同生长阶段的生物被膜形成情况。结果显示,副溶血性弧菌生物被膜的形成是一个动态过程,大概可分为四个阶段:细胞的粘附(12 h)、生物被膜生长(12 h-36 h)、生物被膜成熟(48 h)、生物被膜降解(60 h);副溶血性弧菌在48 h时生物被膜生成量最大。在形成生物被膜的过程中,分泌的胞外物质主要为多糖、蛋白、脂质及核酸等。同时,本研究发现了18个指示特征峰,为探讨与副溶血性弧菌生物被膜形成相关的胞外聚合物的结构与功能提供了理论基础,同时为后续进一步研究复杂的生物被膜系统提供了有力依据。另外,本研究通过拉曼光谱技术首次发现类胡萝卜素在副溶血性弧菌生物被膜形成过程中起协助作用,但其具体调控机制需进一步研究。对副溶血性弧菌生物膜形成过程的深入了解,有助于提高杀菌效率和开发新的和更有效的感染治疗或保护策略。
Raman scattering have recently been proved to be a promising technique for analysing the chemical composition of the extracellular polymer substances(EPS).Microscopic observations such as Confocal laser scanning microscopy(CLSM)and Scanning Electron Microscopy(SEM) have recently been proved to be a promising technique for characterizdng the biofilm matrix.In the present study,the formation of Vibrio parahaemolyticus biofilm on the surface of glass was explained by the first time,and Raman microscopy(RM) analytical technique was used to reveal chemical compositions of EPS in its native state with spatial resolution in μm range.CLAM and SEM were applyed to fully investigate the spatial distribution of Vibrio parahaemolyticus biofilm at different growth phases.The results showed that the formation of biofilm is a dynamic process which includes the following stages:initial reversible attachment,irreversible attachment,bacterial colony formation,mature biofilm development and proliferation,and biofilm dispersion,and the biofilm enhanced according to incubation time and reached a peak at 48 h.The content of carbohydrates,proteins,lipids and nucleic acids in the biofilm matrix increased significantly along with the biofilm growth until 48 h judging from the intensities and appearance probabilities of related marker peaks in the Raman spectra.Moreover,this study found 18 characteristic peaks of EPS.Interestingly,for the first time,carotenoids were found to assist in the formation of the biofilm formation process of V.parahemolyticus.This study indicating that the polysaccharides,proteins,nucleic acids and lipids played an important role in the adhesion,colony aggregation,biofilm maturation and degradation of the bacteria,and providing a theoretical basis for the study of the structure and function of EPS associated with the biofilm formation of V.parahaemolyticus.Elucidation of the basic mechanisms of the biofilm formation could facilitate the deep understanding of basic biochemical mechanisms underlying this process,contribute to enhancing biocide efficiency and developing new and more efficient strategies for the infection treatment or even protection.

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