文章摘要
严羽萍;刘丽;朱军莉;陆海霞;励建荣.鱼源荧光假单胞菌生物被膜形成和致腐表型的研究[J].中国食品学报,2019,19(9):202-209
鱼源荧光假单胞菌生物被膜形成和致腐表型的研究
Studies on Characterization of Biofilm Formation and Spoilage Phenotypes of Pseudomonas fluorescens
  
DOI:
中文关键词: 荧光假单胞菌  生物被膜  腐败  蛋白酶  AHLs
英文关键词: Pseudomonas fluorescens  biofilm  spoilage  protease  AHLs
基金项目:浙江省现代食品安全与营养协同创新中心项目(2017SICR105);浙江省公益技术项目(LGF19C 200003);国家自然科学基金项目(31271954)
作者单位
严羽萍;刘丽;朱军莉;陆海霞;励建荣 浙江工商大学食品与生物工程学院浙江省食品安全重点实验室杭州310018渤海大学辽宁省食品安全重点实验室辽宁锦州121013 
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中文摘要:
      荧光假单胞菌是养殖鱼类低温贮藏中的优势腐败菌。本研究比较分析5种荧光假单胞菌的生物被膜形成和腐败表型。采用结晶紫法、苯酚硫酸法、珠涡流法和荧光显微镜观察荧光假单胞菌的生物被膜形成和粘附能力,并测定细菌泳动性、蛋白酶活性、嗜铁素等致腐表型。结果表明,5株荧光假单胞菌在 28 ℃ LB肉汤中生长良好,经24 h培养后气-液界面上出现较厚的膜,在微孔板中生物被膜形成较快,其中鱼源 PF01、PF06、PF07 和PF10分离株在12 h含量最高,而标准菌株 PFuk4在18 h最高。随着培养时间延长,细菌胞外多糖的含量逐步积累,在18~24 h 达到最高,并较快粘附到不锈钢片表面,其中PF07的粘附量最高。5株荧光假单胞菌还具有较强的泳动性和蛋白酶活性,且均产嗜铁素。在PF07和PFuk4中还检测出短链高丝氨酸内酯(AHLs)活性,可能与其较高的被膜、粘附能力、泳动性及蛋白酶活性有关。本研究结果为从AHLs角度探究荧光假单胞菌的致腐机理打下良好的基础。
英文摘要:
      Pseudomonas fluorescens was dominant spoilage bacteria in cultured fishes during the chilled storage. In the present study, the biofilm formation and spoilage phenotypes of five P. fluorescens isolates were comparatively characterized. The biofilm forming ability and adhesion of the isolates were measured by using crystal violet staining, phenol sulfuric acid assay, bead vortexing method and fluorescence microscopy. Various spoilage phenotypes inculding swimming motility, protease activity, siderophore were performed. The results showed that five P. fluorescens isolates grew fast in LB media at 28 ℃, and formed thick pellicle at the air liquid(A-L) interface for 24 h. All of them produced rapidly biofilm on microtiter plates, and PF01, PF06, PF07 and PF10 reach the maxmium amount of biomass at 12 h, however, PFuk4 arrived at 18 h. The isolates produced continuously exopolysaccharides with the extension of time and arrived to the maxmium amount at 18-24 h. Five isolates adhered to stainless steel quickly, possessed high swimming motility and protease activity, as well as production of siderophore. Only PF07 and PFuk4 were detected to short chain AHLs activity, which could be associated with higher ability of biofilm formation, adhesion, swimming and protease. The present study provided a good fundation to clarify the spoilage mechnism of P. fluorescens based on AHLs.
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