| PCR结合表型鉴定对超高压处理后的冷藏带鱼细菌菌相分析 |
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| 引用本文： | 谢晶,刘骁,杨茜,钱韵芳,张新林.PCR结合表型鉴定对超高压处理后的冷藏带鱼细菌菌相分析[J].农业工程学报,2016,32(5):307-314. |
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| 作者姓名： | 谢晶 刘骁 杨茜 钱韵芳 张新林 |
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| 作者单位： | 上海水产品加工及贮藏工程技术研究中心,上海海洋大学食品学院,上海 201306 |
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| 基金项目： | 国家"十二五"支撑计划课题（2013BAD19B06）;上海市科委重点项目（14dz1205101） |
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| 摘 要： | 为了研究超高压处理对带鱼微生物菌群组成的影响,该文通过形态学特征、生理生化鉴定、16S r RNA序列分析鉴定及系统发育树的建立,分别分析290 MPa、6 min超高压处理前后于4℃冷藏12 d内的带鱼菌相变化,最终分离筛选到24株不同特征的纯化菌株。结果显示,带鱼初始菌相中出现的布式假单胞菌(Pseudomonas brenneri)、黄褐假单胞菌(Pseudomonas fulva)、粪嗜冷杆菌(Psychrobacter faecalis)菌株,经超高压处理后的样品中未能筛选到;波罗的海希瓦氏菌(Shewanella baltica)、隆德假单胞菌(Pseudomonas lundensis)、嗜根寡养单胞菌(Stenotrophomonas rhizophila)、表皮葡萄球菌(Staphylococcus epidermidis)、氧化微杆菌(Microbacterium oxydans)等微生物在超高压处理后的贮藏期间数量逐渐减少至消失;另有一些微生物在贮藏期间逐渐恢复生长,如Rhizobium larrymoorei、Microbacterium halimionae、溶酪大球菌(Macrococcus caseolyticus),而奥斯陆莫拉菌(Moraxella osloensis)、藤黄微球菌(Kocuria rhizophila)、产乳酸菌素的肉杆菌(Carnobacterium maltaromaticum)、西宫皮肤球菌(Dermacoccus nishinomiyaensis)等受超高压的影响较小,尤其是产乳酸菌素的肉杆菌(Carnobacterium maltaromaticum)占好氧菌和厌氧菌菌落总数的比例均较高;Leucobacter aerolatus、成团泛菌(Pantoea agglomerans)、结肠炎耶尔森杆菌palearctica亚种(Yersinia enterocolitica subsp.palearctica)、Chryseobacterium vrystaatense、鼠李糖短杆菌(Brachybacterium rhamnosum)在贮藏末期出现。从带鱼冷藏过程中细菌的组成与变化分析可见,超高压处理对革兰氏阴性菌的抑菌效果较好,而革兰氏阳性菌对超高压处理的耐受性较强。在超高压技术的影响下,致腐败能力较强的微生物被抑制,腐败能力稍弱的微生物成为优势菌,这可能是超高压技术能够有效延长带鱼货架期的因素之一。
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| 关 键 词： | 细菌 冷藏 杀菌 超高压技术 带鱼 聚合酶链式反应(PCR) 生理生化鉴定 菌相 |
| 收稿时间： | 2015/11/23 0:00:00 |
| 修稿时间： | 2015/12/1 0:00:00 |
Microbial ecology of Trichiurus lepturus of haitail pretreated by high hydrostatic pressure during cold storage based on phenotypic and 16S rRNA gene analysis |
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| Xie Jing,Liu Xiao,Yang Xi,Qian Yunfang and Zhang Xinlin.Microbial ecology of Trichiurus lepturus of haitail pretreated by high hydrostatic pressure during cold storage based on phenotypic and 16S rRNA gene analysis[J].Transactions of the Chinese Society of Agricultural Engineering,2016,32(5):307-314. |
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| Authors: | Xie Jing Liu Xiao Yang Xi Qian Yunfang and Zhang Xinlin |
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| Institution: | Shanghai Engineering Research Center of Aquatic Product Processing & Preservation, College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, China,Shanghai Engineering Research Center of Aquatic Product Processing & Preservation, College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, China,Shanghai Engineering Research Center of Aquatic Product Processing & Preservation, College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, China,Shanghai Engineering Research Center of Aquatic Product Processing & Preservation, College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, China and Shanghai Engineering Research Center of Aquatic Product Processing & Preservation, College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, China |
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| Abstract: | The effect of high hydrostatic pressure (290 MPa, 6 min) on microbial ecology changes ofTrichiurus lepturus during vacuum-packaged storage (12 d) was studied via 16S rRNA gene sequencing analysis and phenotypic methods. The microbial colonies of aerobic culture and anaerobic culture, which were obtained from 5 microflora’s growth stages (including freshTrichiurus lepturus, freshTrichiurus lepturus which were stored under 4℃ for 4, 8 and 12 d after high hydrostatic pressure treatment), were isolated and purified by the tryptic soy agar medium and tryptic soy broth medium, repeatedly classified by the traditional phenotypic methods (morphological, physiological and biochemical characteristics of bacteria strains), and then the bacteria DNA of pure culture was extracted and amplified by the polymerase chain reaction (PCR). Finally, 24 bacteria strains of aerobic culture and anaerobic culture in the vacuum-packaged Trichiurus lepturus under the 4℃cold storage were obtained. The result showed thatPseudomonas brenneri,Pseudomonas fulva andPsychrobacter faecalis which appeared in the initial microflora of freshTrichiurus lepturus were inactivated and non-existent after the high hydrostatic pressure treatment; 5 bacteria strains (includingShewanella baltica,Pseudomonas lundensis,Stenotrophomonas rhizophila,Staphylococcus epidermidis andMicrobacterium oxydans) gradually reduced and disappeared after the high hydrostatic pressure treatment. Three bacteria strains gradually grew with the extension of storage time, which were Microbacterium halimionae,Macrococcus caseolyticus andRhizobium larrymoorei. In the whole process of storage after the high hydrostatic pressure treatment, 4 bacteria strains (Moraxella osloensis,Kocuria rhizophila,Carnobacterium maltaromaticum andDermacoccus nishinomiyaensis) always existed, and they were not sensitive to the high hydrostatic pressure treatment. EspeciallyCarnobacterium maltaromaticum, which had a high proportion of total bacteria amount under the conditions of aerobic and anaerobic cultivation, could be considered as a dominate strain during the 4℃ cold storage. At the end of the shelf life ofvacuum-packaged Trichiurus lepturuspretreated by high hydrostatic pressure, 5 bacteria strains (Leucobacter aerolatus,Pantoea agglomerans,Yersinia enterocolitica subsp. Palearctica,Chryseobacterium vrystaatense and Brachybacterium rhamnosum) appeared. The colony evolution ofTrichiurus lepturus during the cold storage showed that Gram negative bacteria were sensitive to the high hydrostatic pressure treatment, while Gram positive bacteria had a high tolerance to the high hydrostatic pressure treatment. Under the influence of high hydrostatic pressure treatment, the microorganisms which have stronger ability of causing corruption are inhibited, and the microorganisms which have weaker ability of causing corruption become dominant bacteria, which may be one of the factors that the high hydrostatic pressure treatment can effectively extend the shelf life ofTrichiurus lepturus. |
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| Keywords: | bacteria cold storage sterilization high hydrostatic pressure treatment Trichiurus lepturus polymerase chain reaction (PCR) physiology-biochemisry technology microflora |
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