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Unveiling hakarl: A study of the microbiota of the traditional Icelandic fermented fish

机译：揭开哈卡尔的面纱：对传统冰岛发酵鱼的微生物群的研究

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摘要

Hakarl is produced by curing of the Greenland shark (Somniosus microcephalus) flesh, which before fermentation is toxic due to the high content of trimethylamine (TMA) or trimethylamine N-oxide (TMAO). Despite its long history of consumption, little knowledge is available on the microbial consortia involved in the fermentation of this fish. In the present study, a polyphasic approach based on both culturing and DNA-based techniques was adopted to gain insight into the microbial species present in ready-to-eat hakarl. To this aim, samples of ready-to-eat hakarl were subjected to viable counting on different selective growth media. The DNA directly extracted from the samples was further subjected to Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis (PCR-DGGE) and 16S amplicon-based sequencing. Moreover, the presence of Shiga toxin-producing Escherichia coli (STEC) and Pseudomonas aeruginosa was assessed via qualitative real-time PCR assays. pH values measured in the analyzed samples ranged from between 8.07 +/- 0.06 and 8.76 +/- 0.00. Viable counts revealed the presence of total mesophilic aerobes, lactic acid bacteria and Pseudomonadaceae. Regarding bacteria, PCR-DGGE analysis highlighted the dominance of close relatives of Tissierella creatinophila. For amplicon sequencing, the main operational taxonomic units (OTUs) shared among the data set were Tissierella, Pseudomonas, Oceanobacillus, Abyssivirga and Lactococcus. The presence of Pseudomonas in the analyzed samples supports the hypothesis of a possible role of this microorganism on the detoxification of shark meat from TMAO or TMA during fermentation. Several minor OTUs ( 1%) were also detected, including Alkalibacterium, Staphylococcus, Proteiniclasticum, Acinetobacter, Erysipelothrix, Anaerobacillus, Ochrobactrum, Listeria and Photobacterium. Analysis of the yeast and filamentous fungi community composition by PCR-DGGE revealed the presence of close relatives of Candida tropicalis, Candida glabrata, Candida parapsilosis, Candida zeylanoides, Saccharomyces cerevisiae, Debaryomyces, Torulaspora, Yamadazyma, Sporobolomyces, Alternaria, Cladosporium tenuissimum, Moristroma quercinum and Phoma/Epicoccum, and some of these species probably play key roles in the development of the sensory qualities of the end product. Finally, qualitative real-time PCR assays revealed the absence of STEC and Pseudomonas aeruginosa in all of the analyzed samples.
机译：Hakarl是由格陵兰鲨鱼（Somniosus microcephalus）的果肉固化产生的，由于高含量的三甲胺（TMA）或三甲胺N-氧化物（TMAO），其发酵前是有毒的。尽管有悠久的消费历史，但有关该鱼发酵的微生物联盟的知识却很少。在本研究中，采用了一种基于培养和基于DNA的技术的多相方法，以深入了解即食哈卡尔酒中存在的微生物种类。为了这个目的，将即食kar豆的样品在不同的选择性生长培养基上进行活计数。直接从样品中提取的DNA进一步进行聚合酶链反应-变性梯度凝胶电泳（PCR-DGGE）和基于16S扩增子的测序。此外，通过定性实时PCR分析评估了产志贺毒素的大肠杆菌（STEC）和铜绿假单胞菌的存在。在分析样品中测得的pH值介于8.07 +/- 0.06和8.76 +/- 0.00之间。可行的计数显示总嗜温需氧菌，乳酸菌和假单胞菌科的存在。关于细菌，PCR-DGGE分析突出显示了产气单胞菌的近亲属的优势。对于扩增子测序，数据集之间共享的主要操作分类单位（OTU）是蒂氏杆菌，假单胞菌，海洋杆菌，阿比西维加和乳球菌。分析样品中假单胞菌的存在支持了这种微生物可能对发酵过程中TMAO或TMA鲨鱼肉排毒的可能作用的假设。还检测到了几个较小的OTU（<1％），包括碱杆菌，葡萄球菌，弹力蛋白菌，不动杆菌属，丹毒菌，厌氧杆菌，O菜，李斯特菌和光细菌。通过PCR-DGGE对酵母和丝状真菌群落组成的分析表明，存在热带假丝酵母，光滑假丝酵母，副念珠菌，玉米假丝酵母，酿酒酵母，德巴利霉菌，龟头孢菌，山梨菌，孢子菌，硬皮孢菌，线粒菌，克罗氏菌的近亲。和Phoma / Epicoccum，其中某些物种可能在最终产品的感官品质发展中起关键作用。最后，定性实时PCR分析显示所有分析样品中均不存在STEC和铜绿假单胞菌。

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  • 来源
    《Food microbiology》 |2019年第9期|560-572|共13页
  • 作者

    Osimani Andrea; Ferrocino Ilario; Agnolucci Monica; Cocolin Luca; Giovannetti Manuela; Cristani Caterina; Palla Michela; Milanovic Vesna; Roncolini Andrea; Sabbatini Riccardo; Garofalo Cristiana; Clementi Francesca; Cardinali Federica; Petruzzelli Annalisa; Gabucci Claudia; Tonucci Franco; Aquilanti Lucia;

  • 作者单位

    Univ Politecn Marche, Dipartimeito Sci Agr Alimentari & Ambientali, Via Brecce Bianche, I-60131 Ancona, Italy;

    Univ Turin, Dept Agr Forest & Food Sci, Largo Paolo Braccini 2, I-10095 Turin, Italy;

    Univ Pisa, Dept Agr Food & Environm, Via Borghetto 80, I-56124 Pisa, Italy|Univ Pisa, Interdept Res Ctr Nutraceut & Food Hlth, Pisa, Italy;

    Univ Turin, Dept Agr Forest & Food Sci, Largo Paolo Braccini 2, I-10095 Turin, Italy;

    Univ Pisa, Dept Agr Food & Environm, Via Borghetto 80, I-56124 Pisa, Italy|Univ Pisa, Interdept Res Ctr Nutraceut & Food Hlth, Pisa, Italy;

    Univ Pisa, E Avanzi Res Ctr, Via Vecchia Marina 6, I-56122 Pisa, Italy;

    Univ Pisa, Dept Agr Food & Environm, Via Borghetto 80, I-56124 Pisa, Italy;

    Univ Politecn Marche, Dipartimeito Sci Agr Alimentari & Ambientali, Via Brecce Bianche, I-60131 Ancona, Italy;

    Univ Politecn Marche, Dipartimeito Sci Agr Alimentari & Ambientali, Via Brecce Bianche, I-60131 Ancona, Italy;

    Univ Politecn Marche, Dipartimeito Sci Agr Alimentari & Ambientali, Via Brecce Bianche, I-60131 Ancona, Italy;

    Univ Politecn Marche, Dipartimeito Sci Agr Alimentari & Ambientali, Via Brecce Bianche, I-60131 Ancona, Italy;

    Univ Politecn Marche, Dipartimeito Sci Agr Alimentari & Ambientali, Via Brecce Bianche, I-60131 Ancona, Italy;

    Univ Politecn Marche, Dipartimeito Sci Agr Alimentari & Ambientali, Via Brecce Bianche, I-60131 Ancona, Italy;

    Ist Zooprofilatt Sperimentale Umbria & Marche, Ctr Riferimento Reg Autocontrollo, Via Canonici 140, I-61100 Villa Fastiggi, Pesaro, Italy;

    Ist Zooprofilatt Sperimentale Umbria & Marche, Ctr Riferimento Reg Autocontrollo, Via Canonici 140, I-61100 Villa Fastiggi, Pesaro, Italy;

    Ist Zooprofilatt Sperimentale Umbria & Marche, Ctr Riferimento Reg Autocontrollo, Via Canonici 140, I-61100 Villa Fastiggi, Pesaro, Italy;

    Univ Politecn Marche, Dipartimeito Sci Agr Alimentari & Ambientali, Via Brecce Bianche, I-60131 Ancona, Italy;

  • 收录信息 美国《科学引文索引》(SCI);美国《化学文摘》(CA);
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    Tissierella; Pseudomonas; Debaryomyces; 16S amplicon-based sequencing; PCR-DGGE;

    机译：Tissierella;假单胞菌;Debaryomyces;基于16S扩增子的测序;PCR-DGGE;

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