食品科学 ›› 2018, Vol. 39 ›› Issue (24): 149-155.doi: 10.7506/spkx1002-6630-201824023

• 生物工程 • 上一篇    下一篇

基于高通量测序的南美白对虾中微生物群落分析

邓晓影,张?宾*,汤?贺,郝桂娟,章样扬   

  1. (浙江海洋大学食品与医药学院,浙江省海产品健康危害因素关键技术研究重点实验室,浙江?舟山 316022)
  • 出版日期:2018-12-25 发布日期:2018-12-17
  • 基金资助:
    中国博士后科学基金项目(2016M590663;2017T100518);浙江省公益性项目(2016C32081); 浙江省自然科学基金项目(LY18C200008)

Analysis of Microbial Diversity in Shrimp (Litopenaeus vannamei) by MiSeq High-Throughput Sequencing

DENG Xiaoying, ZHANG Bin*, TANG He, HAO Guijuan, ZHANG Yangyang   

  1. (Key Laboratory of Health Risk Factors for Seafood of Zhejiang Province, College of Food Science and Pharmacy, Zhejiang Ocean University, Zhoushan 316022, China)
  • Online:2018-12-25 Published:2018-12-17

摘要: 目的:分析不同状态下南美白对虾表面附着的微生物菌群组成情况。方法:以活体对虾、冰鲜对虾、冰鲜虾仁和冷冻虾仁为对象,通过对虾表面附着微生物提取及高通量测序分析,获得不同对虾样品中微生物组成及变化情况。结果:1)各样本有效序列范围为30?591~42?043?条、操作分类单元(operational taxonomic units,OTU)范围为68~168,其中冷冻虾仁OTU数量最低,而活体对虾和冰鲜对虾OTU数量相对较高。2)多样性指数分析发现,所有样品中以冷冻虾仁的ACE、Chao1和Shannon指数值最低、Simpson指数最高,表明其菌群丰度及多样性较低。3)在科水平上,相比于活体对虾,冰鲜对虾中莫拉氏菌科(Moraxellaceae)相对丰度显著增加,其可能与虾体死亡、微生物污染繁殖有关;冰鲜虾仁弧菌科(Vibrionaceae)相对丰度显著增加且含量最高,主要是虾仁加工制备环境及加工器具中微生物接触污染所致;而冷冻虾仁中主要以莫拉氏菌科(Moraxellaceae)、假单胞菌科(Pseudomonadaceae)、李斯特菌科(Listeriaceae)为腐败优势菌。4)热图分析发现,活体对虾和冰鲜对虾菌群结构整体相似度较高,二者与冰鲜虾仁、冷冻虾仁的相似度较低,表明虾仁制备过程、低温冻藏对于虾仁中菌群结构影响较大。5)主成分分析表明,冷冻虾仁与其他组别样本均相距较远,说明冷冻虾仁中微生物种类及组成与其他样品存在着较大差异。结论:南美白对虾样品处于不同状态时,其附着优势微生物、菌群结构、丰度及相互比例等存在显著差异,需针对于不同类型虾类制品,采取不同的控制工艺以保障虾类制品的质量与安全。

关键词: 南美白对虾, 表面微生物, 高通量测序, 菌群分析

Abstract: Objective: To evaluate the composition of microbial communities adhered to the surface of shrimp (Litopenaeus vannamei) under different conditions. Methods: Genomic DNA was extracted from bacteria adhered to the surface of live shrimp, fresh shrimp, fresh peeled shrimp and frozen peeled shrimp and was sequenced by using high-throughput sequencing. Results: 1) A total of 30 591–42 043 valid sequences and 68–168 operational taxonomic units (OTUs) were obtained from each sample, and the number of OTUs was lowest in frozen peeled shrimp and relatively high in live shrimp and fresh shrimp. 2) Among all samples investigated, frozen peeled shrimp exhibited the lowest ACE, Chao1 and Shannon indices and highest Simpson index, suggesting lower bacterial abundance and diversity. 3) At the family level, compared to live shrimp, the relative abundance of Moraxellaceae in fresh shrimp was significantly increased, which might be related to the death of shrimp and microbial contamination and growth; the abundance of Vibrionaceae in fresh peeled shrimp was significantly increased and the highest content was observed, which was caused by microbial contamination from the environment and the devices used during shrimp processing. The dominant spoilage bacteria in frozen peeled shrimp were Moraxellaceae, Pseudomonadaceae, and Listeriaceae. 4) Heatmap analysis showed that the similarity in bacterial flora between live shrimp and fresh shrimp was higher, whereas they were significantly different from fresh peeled shrimp and frozen peeled shrimp, indicating that the preparation of peeled shrimp and frozen storage had a great impact on shrimp bacterial flora. 5) Principal component analysis (PCA) indicated that there was significantly difference in microbial community composition between frozen peeled shrimp and other samples. Conclusion: There were significant differences in the dominant microorganisms, flora structure, abundance and composition among shrimp samples under different conditions. Different control processes are necessary for different types of shrimp products to ensure the quality and safety of shrimp products.

Key words: Litopenaeus vannamei, surface microorganism, high-throughput sequencing, microbial diversity

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