小龙虾(Procambarus clarkii)加工前后优势腐败菌的分离与鉴定  

Isolation and Identification of Specific Spoilage Organisms in Crayfish(Procambarus clarkii)before and after Processing

作　　者：邓灵 赵康 夏开 圣莎丽 解华东[2] 毕旺来 DENG Ling;ZHAO Kang;XIA Kai;SHENG Sha-li;XIE Hua-dong;BI Wang-lai(College of Biological and Pharmaceutical Engineering,Wuhan Polytechnic University,Wuhan 430023,China;Chongqing Academy of Animal Sciences,Chongqing 400039,China)

机构地区：[1]武汉轻工大学生物与制药工程学院,湖北武汉430023 [2]重庆市畜牧科学院,重庆400039

出　　处：《食品工业科技》2020年第18期100-104,共5页Science and Technology of Food Industry

基　　金：重庆市绩效引导资金项目(17427,18231,18508);大学生科研创新项目(xsky2019009)。

摘　　要：以原料小龙虾和卤制加工后的即食小龙虾为对象研究其优势腐败菌种类,为后续研发适宜的杀菌工艺提供理论依据。采用传统平板分离法,从原料小龙虾与开始腐败的即食小龙虾中分别分离出优势腐败菌,通过菌落形态观察、革兰氏染色镜检对微生物种类进行初步分类。采用16S rDNA PCR扩增、限制性内切酶酶切分析以及测序的方法,进行了菌种鉴定试验。结果表明,原料小龙虾优势腐败菌为阴沟肠杆菌(Enterobacter cloacae)、溶酪巨球菌(Macrococcus caseolyticus)、嗜水气单胞菌(Aeromonas hydrophilia)、微小杆菌(Exiguobacterium indicum)和芽胞杆菌属(Bacillus spp.);即食小龙虾优势腐败菌仅检测到以蜡样芽孢杆菌(Bacillus cereus)与苏云金芽孢杆菌(Bacillus thuringiensis)为代表的芽孢杆菌,而其他类型的细菌在卤制加工过程中已被灭活。小龙虾加工前后优势腐败菌鉴定结果对比分析可知,高温卤制工艺难以除灭原料小龙虾中的芽孢杆菌,芽孢杆菌仍存活导致即食小龙虾腐败。In order to explore the suitable sterilizing process,the main types of specific spoilage organisms in crayfish before and after processing were investigated.The spoilage bacteria were isolated from unprocessed raw crayfish and marinated ready-to-eat crayfish by using traditional plate separation method.The bacteria species were initially classified by colony morphological observation and Gram staining.Then 16S rDNA PCR,restriction endonuclease digestion analysis and 16S rDNA sequencing were performed to identify the screened bacteria.The specific spoilage organisms from raw crayfish were determined to be Macrococcus caseolyticus,Aeromonas hydrophilic,Enterobacter cloacae,Exiguobacterium indicum and Bacillus spp.Compared with raw crayfish,only Bacillus spp.including Bacillus cereus and Bacillus thuringiensis were detected in ready-to-eat crayfish,as other types of spoilage organisms were killed after marinating operation.The results showed that Bacillus spp.still survived in ready-to-eat crayfish after processing,thus causing corruption.

关 键 词：小龙虾 优势腐败菌 菌种鉴定 16S rDNA测序 限制性内切酶消化 

分 类 号：TS254.1[轻工技术与工程—水产品加工及贮藏工程]