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PCR-DGGE法结合传统技术研究4℃冷链贮运条件下三文鱼菌相变化
引用本文:张新林,谢 晶,钱韻芳,刘永吉,曾丹妮.PCR-DGGE法结合传统技术研究4℃冷链贮运条件下三文鱼菌相变化[J].食品科学,2016,37(24):271-277.
作者姓名:张新林  谢 晶  钱韻芳  刘永吉  曾丹妮
作者单位: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,上海 201306
基金项目:国家自然科学基金面上项目(31571914);2016年上海市科技兴农重点攻关项目(沪农科攻字(2016)第1-1号);上海市科委研究中心能力提升项目(16DZ2280300)
摘    要:为研究三文鱼在冷链贮运4℃条件下的细菌腐败机制,运用聚合酶链式反应-变性梯度凝胶电泳(polymerase chain reaction-denaturing gradient gel electrophoresis,PCR-DGGE)技术、传统鉴定技术以及PCR技术分析了4℃冷链贮运条件下的三文鱼中腐败菌菌相变化规律,并通过产量因子测得各优势菌株致腐能力,从而确定特定腐败菌。DGGE指纹图谱显示,贮藏期间微生物多样性降低,假单胞菌属和希瓦氏菌属条带亮度却逐渐提高。这表明这两个属的微生物在三文鱼冷藏期间逐渐成为优势菌。通过分离和鉴定贮藏末期腐败三文鱼的优势腐败菌,本实验得到5株优势腐败菌,分别为麦芽糖肉食杆菌(Carnobacterium maltaromaticum LMA28)、丁香假单胞菌(Pseudomonas syringae pv.syringae B728a)、荧光假单胞菌(Pseudomonas fluorescens SBW25)、肉食杆菌(Carnobacterium sp.WN1359)和波罗的海希瓦氏菌(Shewanella baltica OS678)。将这5株纯培养的腐败菌分别接种到无菌三文鱼中并冷藏一定时间后,各腐败菌的挥发性盐基氮(total volatile basic nitrogen,TVB-N)产量因子分别为1.26×10~(-7)、1.25×10~(-7)、1.36×10~(-7)、1.08×10~(-7)mg TVB-N/CFU和1.03×10~(-7)mg TVB-N/CFU。这5株腐败菌对三文鱼致腐败能力的顺序依次为荧光假单胞菌SBW25麦芽糖肉食杆菌LMA28丁香假单胞菌B728a肉食杆菌WN1359波罗的海希瓦氏菌OS678。

关 键 词:三文鱼  菌相变化  特定腐败菌  荧光假单胞  致腐能力  

Detection of Microfloral Changes in Salmon during Cold Chain Storage and Transport at 4℃ by PCR-DGGE and Phenotypic Analysis
ZHANG Xinlin,XIE Jing,QIAN Yunfang,LIU Yongji,ZENG Danni.Detection of Microfloral Changes in Salmon during Cold Chain Storage and Transport at 4℃ by PCR-DGGE and Phenotypic Analysis[J].Food Science,2016,37(24):271-277.
Authors:ZHANG Xinlin  XIE Jing  QIAN Yunfang  LIU Yongji  ZENG Danni
Affiliation:Shanghai Engineering Research Center of Aquatic Product Processing & Preservation, College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, China
Abstract:This study attempted to elucidate the bacterial spoilage mechanism of salmon during cold chain storage and transport at 4 ℃. Polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE), the traditional identification method and PCR were adopted to analyze the microflora changes in salmon during cold chain storage and transport at 4 ℃ and identify the specific spoilage organisms by testing the spoilage abilities of the dominant strains with yield factors. DGGE fingerprint showed that the diversity of bacterial species decreased during the cold storage of salmon, but the brightness of bands for Pseudomonas spp. and Shewanella spp. increased, which indicated that both strains became the predominant bacteria during the storage. Five dominant spoilage bacteria, Carnobacterium maltaromaticum LMA28, Pseudomonas syringae pv. syringae B728a, Pseudomonas fluorescens SBW25, Carnobacterium sp. WN1359 and Shewanella baltica OS678, were isolated and identified at the end of storage. After separately inoculating the 5 dominate spoilage organisms into sterilized salmon and refrigerating them for a certain period of time, their yield factors of TVB-N were 1.26 × 10-7, 1.25 × 10-7, 1.36 × 10-7, 1.08 × 10-7 and 1.03 × 10-7 mg TVB-N/CFU, respectively. These results showed that P. fluorescens SBW25 has the strongest spoilage ability for salmon, followed by C. maltaromaticum LMA28, P. syringae pv. syringae B728a, Carnobacterium sp. WN1359 and S. baltica OS678 successively.
Keywords:salmon  microfloral changes  special spoilage organisms (SSOs)  Pseudomonas fluorescens  spoilage ability  
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