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Novel insights into the enterotoxigenic potential and genomic background of Staphylococcus aureus isolated from raw milk

机译：从生牛奶中分离出肠毒素潜力和葡萄球菌基因组背景的新颖见解

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摘要

In this study, 53 Staphylococcus (S.) aureus strains were typed by 16S-23S rDNA intergenic spacer region (ISR) typing and staphylococcal enterotoxin gene (SEg) typing for all the staphylococcal enterotoxin (se) and sta-phylococcal enterotoxin-like toxin (sel) genes known to date, revealing a higher discriminatory power than that of multi locus sequence typing. Six strains, one of each ISR- and SEg-type, were genome sequenced and the ability to produce some classical and new SEs when growing in milk was investigated. The manual analysis of the six genomes allowed us to confirm, correct and expand the results of common available genomic data pipelines such as VirulenceFinder. Moreover, it enabled us to (ⅰ) investigate the actual location of se and sel genes, even for genes such as selY, whose location (in the core genome) was so far unknown, (ⅱ) find novel allelic variants of se and sel genes and pseudogenes, (ⅲ) correctly annotate se and sel genes and pseudogenes, and (ⅳ) discover a novel type of enterotoxin gene cluster (egc), i.e. the egc type 5 in strains 356P and 364P, while S. argenteus MSHR1132 harbored the egc type 6. Four of the six S. aureus strains produced sufficient amounts of SEA, SEC, SED and SEH in milk to cause staphylococcal food poisoning (SFP), with S. aureus 372 P being the highest producer of SED in milk found to date, producing as much as ca. 47,300 ng/mL and 49,200 ng/mL of SED, after 24 and 48 h of incubation in milk at 37 °C, respectively. S. aureus 372 P released a low amount of SER in milk, most likely because the seR gene was present as a pseudogene, putatively encoding only 51 amino acids. These findings confirm that not only the classical SEs, but also the new ones can represent a potential hazard for the consumers' health if produced in foods in sufficient amounts. Therefore, the detection of SEs in foods, especially if involved in SFP cases, should focus not only on classical, but also on all the new SEs and SEls known to date. Where reference methods are unavailable, the presence of the relevant genes, by using the conventional and real time PCR protocols we exhaustively provided herein, and their nucleotide sequences, should be investigated.
机译：在本研究中，53个葡萄球菌菌株串行于16s-23s rdNA代骨间隔区（ISR）键入和葡萄球菌肠毒素基因（SEG），其键入所有葡萄球菌肠毒素（SE）和STA-Phylococal Enterotoxin样毒素（SEL）迄今为止所知的基因，揭示了比多基因座序列序列的歧化功率更高。六种菌株，每种ISR-和SEG型，是基因组测序，并在调查牛奶中生长时产生一些经典和新的SE的能力。六种基因组的手动分析使我们能够确认，纠正和扩展常见的基因组数据管道的结果，如毒力Finder。此外，它使我们能够研究SE和SEL基因的实际位置，即使是基因，甚至是其位置（在核心基因组）的位置到目前为止，（Ⅱ）发现SE和SEL的新型等位基因变种基因和假生素，（Ⅲ）正确注释Se和Sel基因和假性引发，并且（ⅳ）发现一种新型的肠毒素基因簇（EGC），即菌株356P和364P中的EGC型5，而S. Argenteus MSHR1133则陷入困境EGC型6.六个金黄色葡萄球菌中的四种菌株产生足够量的牛奶中的海，SEC，SED和SEH，以引起葡萄球菌食物中毒（SFP），用S.UUREUS 372 P是牛奶中的最高生产者日期，制造尽可能多。 47,300ng / ml和49,200ng / ml sed，24和48小时后，分别在37°C的牛奶中孵育。 S. aureus 372p释放了少量的牛奶，最有可能因为Ser基因作为假基因存在，仅借助于51个氨基酸。这些调查结果证实，不仅是经典的SE，而且新的结果可能代表消费者健康的潜在危害，如果在足够的食物中生产。因此，在食品中检测SES，特别是如果参与SFP案件，则应不仅关注古典，而且还关注迄今为止所知道的所有新的SES和SEL。如果参考方法不可用，则应研究通过使用常规和实时PCR方案的相关基因的存在，以及我们彻底地提供的，以及它们的核苷酸序列，以及它们的核苷酸序列。

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  • 来源
    《Food microbiology》 |2020年第9期|103482.1-103482.16|共16页
  • 作者

    Daniele Chieffi; Francesca Fanelli; Gyu-Sung Cho; Justyna Schubert; Giuseppe Blaiotta; Charles M.A.P. Franz; Jacek Bania; Vincenzina Fusco;

  • 作者单位

    National Research Council of Italy Institute of Sciences of Food Production (CNR-ISPA) Bari Italy;

    National Research Council of Italy Institute of Sciences of Food Production (CNR-ISPA) Bari Italy;

    Max Rubner-Institut Department of Microbiology and Biotechnology Hermann-Weigmann-Strasse 1 24103 Kiel Germany;

    Department of Food Hygiene and Consumer Health Protection Wroclaw University of Environmental and Life Sciences Wroclaw Poland;

    Department of Agricultural Sciences Division of Vine and Wine Sciences University of Naples Federico Ⅱ Viale Italia 83100 Avellino Italy;

    Max Rubner-Institut Department of Microbiology and Biotechnology Hermann-Weigmann-Strasse 1 24103 Kiel Germany;

    Department of Food Hygiene and Consumer Health Protection Wroclaw University of Environmental and Life Sciences Wroclaw Poland;

    National Research Council of Italy Institute of Sciences of Food Production (CNR-ISPA) Bari 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

    Staphylococcus aureus; Staphylococcus argenteus; Staphylococcal enterotoxin; Enterotoxin gene cluster; Staphylococcus aureus pathogenicity Island(SaPI); Raw milk;

    机译：金黄色葡萄球菌;葡萄球菌argenteus;葡萄球菌肠毒素;肠毒素基因簇;金黄色葡萄球菌致病岛（SAPI）;生牛奶;

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