Abstract

Currently, only 5 (SEA to SEE) out of 27 known staphylococcal enterotoxins can be analyzed using commercially available kits. Six genes (seg, sei, sem, sen, seo, and seu), encoding putative and undetectable enterotoxins, are located on the enterotoxin gene cluster (egc), which is part of the Staphylococcus aureus genomic island vSaβ. These enterotoxins have been described as likely being involved in staphylococcal food-poisoning outbreaks. The aim of the present study was to determine if whole-genome data can be used for the prediction of staphylococcal egc enterotoxin production, particularly enterotoxin G (SEG) and enterotoxin I (SEI). For this purpose, whole-genome sequences of 75 Staphylococcus aureus strains from different origins (food-poisoning outbreaks, human, and animal) were investigated by applying bioinformatics methods (phylogenetic analysis using the core genome and different alignments). SEG and SEI expression was tested in vitro using a sandwich enzyme-linked immunosorbent assay method. Strains could be allocated to 14 different vSaβ types, each type being associated with a single clonal complex (CC). In addition, the vSaβ type and CC were associated with the origin of the strain (human or cattle derived). The amount of SEG and SEI produced also correlated with the vSaβ type and the CC of a strain. The present results show promising indications that the in vitro production of SEG and SEI can be predicted based on the vSaβ type or CC of a strain. IMPORTANCE Besides having infectious properties in human and animals, S. aureus can produce different enterotoxins in food. The enterotoxins can cause vomiting and diarrhea, often involving many people. Most of these outbreaks remain undiscovered, as detection methods for enterotoxins are only available for a few enterotoxins but not for the more recently discovered enterotoxins G (SEG) and I (SEI). In this study, we show promising results that in vitro production of SEG and SEI can be predicted based on the whole-genome sequencing data of a strain. In addition, these data could be used to find the source (human or cattle derived) of an outbreak strain, which is the key for a better understanding of the role SEG and SEI play in foodborne outbreaks caused by S. aureus.

Highlights

  • Only 5 (SEA to SEE) out of 27 known staphylococcal enterotoxins can be analyzed using commercially available kits

  • Multilocus sequence typing (MLST) of the 75 S. aureus strains isolated from different sources, like food, humans, animals, and the environment, showed that the most frequently found clonal complexes (CC) are CC5 (n = 17), CC20 (n = 15), CC30 (n = 13), and CC705 (n = 11), followed by CC45, CC22, CC50, and CC9 (6, 3, 2, and 2 strains, respectively)

  • For 15 strains, spa typing resulted in an unknown type, of which the majority belonged to CC30 and CC20 (5 and 6 unknown spa types, respectively)

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Summary

Introduction

Only 5 (SEA to SEE) out of 27 known staphylococcal enterotoxins can be analyzed using commercially available kits. The aim of the present study was to determine if whole-genome data can be used for the prediction of staphylococcal egc enterotoxin production, enterotoxin G (SEG) and enterotoxin I (SEI) For this purpose, whole-genome sequences of 75 Staphylococcus aureus strains from different origins (food-poisoning outbreaks, human, and animal) were investigated by applying bioinformatics methods (phylogenetic analysis using the core genome and different alignments). We show promising results that in vitro production of SEG and SEI can be predicted based on the whole-genome sequencing data of a strain These data could be used to find the source (human or cattle derived) of an outbreak strain, which is the key for a better understanding of the role SEG and SEI play in foodborne outbreaks caused by S. aureus. Literature suggests that about 50% of S. aureus strains harbor an egc [21, 23, 24]

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