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1 5 7 Vol. 5 No Journal of Food Safety and Quality Jul., 2014 熊振海, 马晨晨 * (, ) 摘要 : 目的,, 方法 16S rdna 结果 CFU/g, 8, : S01 S02, S03, S04 S06, S05, S07, S08 结论, 4, 5 关键词 : ; ; ; 16S rdna Analysis of cooling beef shelf life and microbial diversity XIONG Zhen-Hai, MA Chen-Chen * (Shanghai Engineering Research Center of Aquatic-Product Processing & Preservation, College of Food Science and Technology, Shanghai Ocean University, Shanghai , China) ABSTRACT: Objective To provide theoretical data for the storage of chilled beef by the initial microbial in the chilled beef and understanding the structure of bacterial colonies. Methods Combined the traditional pure culture methods and the analysis of 16S rdna clone to study the total number of colonies of microorganisms and structure in chilled beef. Results The total number of bacteria in chilled beef was CFU/g, and screening eight dominant strains in it. Identifying by their morphology, color and other characteristics, confirm that it had the following: S01 and S02 were Pseudomonas spp., S03 was Shewanella, S04, S06 was Acinetobacter, S05 was narrow food Aeromonas spp., S07 was psychrophilic bacterium, and S08 was Staphylococcus spp.. Conclusion Pseudomonas and Shewanella are predominant bacterium in the chilled beef, the shelf life of the chilled beef was 5 days at 4 conditions. KEY WORDS: chilled beef; microbial diversity; total number of colonies; 16S rdna 1 引言,, [1],, [2],,, 基金项目 : (B ) Fund: Supported by the Project Construction of Laboratory Technical Team(B ) * 通讯作者 :,, ccma@shou.edu.cn *Corresponding author: MA Chen-Chen, Master, Laboratory Assistant, College of Food Science and Technology, Shanghai Ocean University,No.999, Hucheng Huan Road, Pudong District, Shanghai , China. ccma@shou.edu.cn

2 2110 5, 10 8 CFU/cm 2,,, [3],,,, [4,5],,, [6],,,,, [7-9],,,, 2 材料与方法 2.1 材料与试剂 ( ), ( 25 g),, 4, 4 d 12 h, 5 d 24 h ; PCR ( ); Biospin DNA ( ); ( ) 2.2 仪器与设备 ( ); DHG-9140A ( ); AUY120 ( ); ( ); SWCJCO ( ); NikonE200 ( ( ) ); PCR ( Eppendorf); ( UVP ); ( Bio-Rad ); 2.3 方法 冷却牛肉菌落总数测定 10-8, 10-1 ~ ml, NA, 37±1 48 h 3, 冷却牛肉初始菌体的培养与分离纯化,, 24~48 h,,, 经纯化的初始单菌落 DNA 提取及纯化 Biospin DNA ( ), DNA SrDNA 的 PCR 扩增 DNA, : 1492r: 5 -CTACGGCTACCTTG- TTAC GA-3, 27f: 5 -GAGA GTTTGAT- CCTGGCTCAG-3, PCR 25 μl PCR : 95, 3 min, 95, 1 min, 55, 1 min, 72 1 min, 25, 72, 10 min DNA PCR DNA 测序 ( 1400 bp) 16S rdna, Blast GenBank ( Nih.gov/blast) 3 结果与分析 3.1 冷却牛肉初始菌落及变化规律, : CFU/g, 1 :

3 7, : 2111, 1500 kb 3.4 结果 Blast GenBank, : S01 S02, S03, S04 S06, S05, S07, S08 1 Fig. 1 Total beef colonies cooling curve 2 d, 8 d 3.2 细菌菌落特征, 17,,,, 8 (8 : S01-S08),, DNA 的提取和 PCR 扩增 2, 8 DNA, 1%, 23 kb, DNA 3, 8 16S PCR, 4 结论,,, 10 7 CFU/g,, 10 8 CFU/g, 10 6 CFU/g, 4, CFU/g, 4 6, 8, S01 S02, S03, S04 S06, S05, S07, S08, [9-12] [13] SSO(specific spoilage organisms) Table 1 表 1 冷却牛肉中细菌特征 The properties of bacteria on chilled beef S01 S02 S03 S04 S05 S06 S07 S08 G- G- G- G- G- G+ G+ G-

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5 7, : 2113 [J]., 2009, 32(2): Li ZT, Li BL, Ou J. Screening and Identification of the Primary Bacterium from the Chilled Beef on Sale[J]. Microbiol, 2009, Hu P, Zhou GH, Xu XL, et al. Study of the dynamic changes of microorganisms in sliced vacuum-packed cooked ham during storage by PCR-DGGE fingerprinting[j]. J Nanjing Agric Univ, 2009, 32(2): [13],,. PCR-DGGE 36(2): 作者简介 ( 责任编辑 : 邓伟 ) [J]., 2008, 36(9): Li MY, Zhou GH, Xu XL. Study on the dominant bacteria during the storage period on chilled pork using PCR-DGGE [J]. J 熊振海, 副教授, 主要研究方向为食品安全信息系统开发, 食品质量与安全追溯及风险预警 计算化学 zhxiong@shou.edu.cn Northwest A F University(Nat Sci Edi), 2008, 36(9): [14] Gill CO, Badoni M, Jones T. Hygienic effects of trimming and washing operations in a beef carcass dressing process [J]. J Food Prot, 1996, 59: 马晨晨, 工学硕士, 主要研究方向为食品微生物 ccma@shou.edu.cn [15],,,. [J]., 2009, 36(2):