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

被引:39
|
作者
Osimani, Andrea [1 ]
Ferrocino, Ilario [2 ]
Agnolucci, Monica [3 ,4 ]
Cocolin, Luca [2 ]
Giovannetti, Manuela [3 ,4 ]
Cristani, Caterina [5 ]
Palla, Michela [3 ]
Milanovic, Vesna [1 ]
Roncolini, Andrea [1 ]
Sabbatini, Riccardo [1 ]
Garofalo, Cristiana [1 ]
Clementi, Francesca [1 ]
Cardinali, Federica [1 ]
Petruzzelli, Annalisa [6 ]
Gabucci, Claudia [6 ]
Tonucci, Franco [6 ]
Aquilanti, Lucia [1 ]
机构
[1] Univ Politecn Marche, Dipartimeito Sci Agr Alimentari & Ambientali, Via Brecce Bianche, I-60131 Ancona, Italy
[2] Univ Turin, Dept Agr Forest & Food Sci, Largo Paolo Braccini 2, I-10095 Turin, Italy
[3] Univ Pisa, Dept Agr Food & Environm, Via Borghetto 80, I-56124 Pisa, Italy
[4] Univ Pisa, Interdept Res Ctr Nutraceut & Food Hlth, Pisa, Italy
[5] Univ Pisa, E Avanzi Res Ctr, Via Vecchia Marina 6, I-56122 Pisa, Italy
[6] Ist Zooprofilatt Sperimentale Umbria & Marche, Ctr Riferimento Reg Autocontrollo, Via Canonici 140, I-61100 Villa Fastiggi, Pesaro, Italy
关键词
Tissierella; Pseudomonas; Debaryomyces; 16S amplicon-based sequencing; PCR-DGGE; LACTIC-ACID BACTERIA; GEN; NOV; TRIMETHYLAMINE OXIDE; LACTOCOCCUS-LACTIS; PCR-DGGE; YEASTS; DIVERSITY; SEAFOOD; IDENTIFICATION; COMMUNITY;
D O I
10.1016/j.fm.2019.03.027
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
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.
引用
收藏
页码:560 / 572
页数:13
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