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利用非培养 16S rDNA 序列分析鉴定冷藏过程中变质脆化草鱼(Ctenopharyngodon idellus C. et V.)和草鱼(Ctenopharyngodon idellus)鱼片的优势微生物种属。

Identification of the Dominant Microbial Species of Spoiled Crisp Grass Carp ( Ctenopharyngodon idellus C. et V.) and Grass Carp ( Ctenopharyngodon idellus) Fillets during Cold Storage by Culture-Independent 16S rDNA Sequence Analysis.

机构信息

University of Electronic Science and Technology of China, Zhongshan Institute, Zhongshan, 528402, People's Republic of China.

出版信息

J Food Prot. 2018 Jan;81(1):84-92. doi: 10.4315/0362-028X.JFP-17-141.

Abstract

A new freshwater cultivation species, crisp grass carp (CGC; Ctenopharyngodon idellus C. et V.) has a special texture and is popular with consumers; thus, we should pay close attention to its storage conditions and bacterial degradation. CGC and grass carp (GC; Ctenopharyngodon idellus) were commercialized as fillets and subsequently stored at 4 and 8°C. Microbial growth parameters (total viable counts, psychrotrophic bacteria, and Pseudomonas spp.), physicochemical data (pH and total volatile base nitrogen), and sensory analysis were monitored during the storage period. Dominant microorganisms were identified using a 16S rDNA clone library and restriction fragment length polymorphism (RFLP) analysis after the fillets had spoiled completely. The results showed that Pseudomonas spp. lagged behind the psychrotrophic population and the total viable counts initially and increased more rapidly after storage for 2 days. Total volatile base nitrogen contents were found to increase with storage time in both species, coinciding with ongoing microbial change. The pH results obtained for both species during storage showed an overall increase at the end of storage. Sensory evaluation showed a shelf life of 3 and 6 days for both species at 8 and 4°C, respectively. RFLP analysis of the 16S rDNA clone library revealed that there were seven and five distinct RFLP pattern groups in the microflora of the spoiled CGC and GC fillets, respectively. Through RFLP patterns and 16S rDNA sequencing from the clones, it was determined that CGC fillets stored at 4°C were dominated by Pseudomonas spp. at the point of sensory rejection, whereas GC fillets were dominated by populations affiliated with Pseudomonas sp., Acinetobacter sp., and Aeromonas sp.

摘要

一种新的淡水养殖品种脆肉鲩(CGC;草鱼 C. et V.)具有特殊的质地,深受消费者欢迎;因此,我们应该密切关注其储存条件和细菌降解。CGC 和草鱼(GC;草鱼)被商业化制成鱼片,然后分别在 4°C 和 8°C 下储存。在储存期间监测微生物生长参数(总活菌数、嗜冷菌和假单胞菌)、理化数据(pH 值和总挥发性碱基氮)和感官分析。在鱼片完全变质后,使用 16S rDNA 克隆文库和限制性片段长度多态性(RFLP)分析鉴定优势微生物。结果表明,假单胞菌滞后于初始的嗜冷菌群和总活菌数,在储存 2 天后增加得更快。两种鱼的总挥发性碱基氮含量均随储存时间的延长而增加,与微生物的持续变化相吻合。两种鱼在储存过程中的 pH 值结果表明,在储存结束时总体呈上升趋势。感官评价表明,在 8°C 和 4°C 下,两种鱼的货架期分别为 3 天和 6 天。16S rDNA 克隆文库的 RFLP 分析表明,变质的 CGC 和 GC 鱼片的微生物群中分别有 7 个和 5 个不同的 RFLP 模式群。通过 RFLP 模式和克隆的 16S rDNA 测序,确定在感官接受点,4°C 储存的 CGC 鱼片主要由假单胞菌属,而 GC 鱼片主要由假单胞菌属、不动杆菌属和气单胞菌属组成。

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