A comprehensive review on the detection of Staphylococcus aureus enterotoxins in food samples.

Staphylococcal enterotoxins (SEs), the major virulence factors of Staphylococcus aureus, cause a wide range of food poisoning and seriously threaten human health by infiltrating the food supply chain at different phases of manufacture, processes, distribution, and market. The significant prevalence of Staphylococcus aureus calls for efficient, fast, and sensitive methods for the early detection of SEs. Here, we provide a comprehensive review of the hazards of SEs in contaminated food, the characteristic and worldwide regulations of SEs, and various detection methods for SEs with extensive comparison and discussion of benefits and drawbacks, mainly including biological detection, genetic detection, and mass spectrometry detection and biosensors. We highlight the biosensors for the screening purpose of SEs, which are classified according to different recognition elements such as antibodies, aptamers, molecularly imprinted polymers, T-cell receptors, and transducers such as optical, electrochemical, and piezoelectric biosensors. We analyzed challenges of biosensors for the monitoring of SEs and conclude the trends for the development of novel biosensors should pay attention to improve samples pretreatment efficiency, employ innovative nanomaterials, and develop portable instruments. This review provides new information and insightful commentary, important to the development and innovation of further detection methods for SEs in food samples.

Effect of environmental factors on expression of staphylococcal enterotoxin genes.

Staphylococcal enterotoxins (SEs) secreted by Staphylococcus aureus (S. aureus) can cause foodborne disease, nausea, vomiting and diarrhea, and even death. Regulation of SE expression is related to accessory gene regulators (Agr). It is important to reveal which environmental factors influence regulation of SE expression to prevent SE food poisoning outbreak. Hence, natural environmental factors which may have an impact on SE expression were selected, such as temperature, food types, strains, and competing strains. Seven strains of S. aureus carrying different SE genes were collected from the Chinese Academy of Inspection and Quarantine (CAIQ) strain bank for study. Strains were cultured with different conditions. Temperature was 8 °C, 22 °C, and 30 °C. Food type was milk powder and nutrient broth. Competing strains were Vibrio parahaemolyticus (V. parahaemolyticus), Escherichia coli (E. coli), and Bacillus cereus (B. cereus). The expression culture solution was pretreated by centrifugation, then determined by using SDS-PAGE, and distinguished SEs apart from each other by HPLC-ESI-TOF. There are 168 samples collected from SE expression culture; the result of SDS-PAGE suggests 23 samples were positive for SEs, and the other 145 samples were negative for SEs. The result of HPLC-ESI-TOF suggests that SEs with similar molecular weight can be distinguished in terms of m/z. The most important factor contributing to regulate expression of SEs was estimated by logistic regressive analysis. The result shows that McFadden R2 is 0.213; p value is 0.000 (p < 0.05); this result illustrates that the model is valid and meaningful. Strains, food types, temperature, and competing strands can explain the 21% change in SE expression. Temperature (z = 3.029, p = 0.002 < 0.01), strains (z = - 3.132, p = 0.002 < 0.01), and food types (z = - 2.415, p = 0.016 < 0.05) have significant impact on SE expression, and the competing strains (z = 1.230, p = 0.219 > 0.05) have no impact on the SE expression. More important impact on SE expression was estimated by OR value; the result shows that strength of temperature influencing on SE expression is bigger than strains and food types in terms of values of OR, temperature (OR = 2.862), strains (OR = 0.641), and food types (OR = 0.561); consequently, temperature is a key factor for stimulating SE expression and had high expression at 30 °C. Therefore, food easily contaminated with S. aureus should be monitored intensively at early and late summer, when proper temperature for expressing SEs may result in S. aureus food poisoning prevalence.

Epidemiological and molecular evidence of foodborne poisoning outbreak caused by enterotoxin gene cluster-harboring Staphylococcus aureus of new sequence type 7591.

OBJECTIVES: This paper presented a detailed analysis of the epidemiology and molecular characteristics of staphylococcal food poisoning (SFP) that occurred in a hotel in Hangzhou. METHODS: A total of 46 guests at the hotel underwent an epidemiological survey. Samples of stool from patients, vomit, swabs from the kitchen, leftover food items, and anal swabs from food handlers were taken and investigated for the presence of potential pathogenic bacteria. Molecular techniques and whole genome sequencing were performed to track the evolution of Staphylococcus aureus associated with the outbreak of SFP. RESULTS: Forty-six individuals displayed gastrointestinal symptoms. Seventeen isolates of S. aureus were discovered to carry the seg, sei, sem, sen, seo, and selu genes found in a specific enterotoxin gene cluster (egc) operon, but without the presence of classical enterotoxins such as SEA ∼ SEE. All egc-positive isolates shared identical pulsed-field gel electrophoresis profiles and were classified under new ST7591 (Clonal Complex 72) with identical spa typing t148. In addition, some isolates of S. aureus obtained from food sources sold in Hangzhou over the past 3 years and carrying egc genes were grouped under the ST72 lineage (CC72). Through whole genome sequencing, a strong genetic connection was revealed between these egc-positive isolates and clinical ST72 S. aureus found in China. CONCLUSIONS: S. aureus with non-classical egc enterotoxins was suggested to be a potential cause of SFP in humans.

Investigation of a Staphylococcus aureus sequence type 72 food poisoning outbreak associated with food-handler contamination in Italy.

On August 2019 a staphylococcal food poisoning outbreak occurred in an elderly home in Piedmont, Italy. The epidemiological investigation performed among the persons that consumed the meal identified chicken salad as the most likely source of the outbreak. Staphylococcus aureus was isolated from a total of seven samples, namely one vomit sample from a guest of the nursing home, two food samples (chicken salad with and without mayonnaise) and nasal swabs collected from a total of four persons working in the kitchen of the nursing home. The maximum likelihood tree obtained using single nucleotide polymorphisms analysis revealed that the isolates from the aforementioned samples clustered together. Multilocus sequence typing revealed that they belonged to Sequence Type 72. Fourier transform infrared spectroscopy (FTIR) was used in parallel to single nucleotide polymorphisms and whole genome sequencing for the determination of the degree of relatedness of the isolates. The results of the FTIR showed the same clustering obtained with single nucleotide polymorphisms and whole genome sequencing and revealed the source of infection. This study underlines the importance of both laboratory evidence and epidemiological data for outbreak investigation and further confirms that FTIR is a suitable support for the short-term epidemiological investigation on source attribution in case of a S. aureus infection.

First report of enterotoxigenic Staphylococcus argenteus as a foodborne pathogen.

Staphylococcal enterotoxins preformed in food are the causative agents of staphylococcal food poisoning outbreaks (SFPO). In this study we characterised in depth two coagulase-positive non-pigmented staphylococci involved in two independent outbreaks that occurred in France. While indistinguishable from Staphylococcus aureus using PCR methods and growth phenotype comparisons, both isolates were identified as Staphylococcus argenteus by whole genome sequencing. The genomes were analysed for the presence of enterotoxin genes, whose expression was determined in laboratory medium and, for the first time, in artificially-contaminated milk samples by using liquid chromatography-mass spectrometry and ELISA methods. The concentration measured for the SEB toxin in milk (0.67 ng/ml) was comparable to concentrations reported for other types of enterotoxins behind SFPO. From a collection of publicly available genomes, we performed an unprecedented systematic investigation of the enterotoxin gene set of S. argenteus, including variants and pseudogenes. The most prevalent genes were sex, followed by sel26, sel27 and sey. The egc cluster was less frequent and most of the time carried a dysfunctional seg gene. Our results shed light on the enterotoxigenic properties of S. argenteus, and emphasize the importance in monitoring of S. argenteus as an emerging foodborne pathogen.

Prevalence of staphylococcal toxin in food contaminated by Staphylococcus spp.: Protocol for a systematic review with meta-analysis.

BACKGROUND: Food contamination by Staphylococcus spp. enterotoxigenic strains is quite common and despite underreporting caused by the short duration of clinical symptoms and lack of medical care, staphylococcal food poisoning is one of the most common Foodborne Diseases (FBD) in the world. This study describes a systematic review protocol with meta-analysis on the prevalence and types of staphylococcal enterotoxins in food, and the profile of contaminated foods. METHODS: The research will be conducted through the selection of studies reporting the analysis of staphylococcal enterotoxins in food contaminated by Staphylococcus spp. Searches will happen on the following databases: Medline (OVID), GALE, Science Direct, CAB Direct (CABI), Google Scholar, in addition to manual search in the list of references of articles, directory of theses and dissertations, and countries' health agencies. Reports will be imported into the application Rayyan. Two researchers will independently select studies and extract data, and a third reviewer will solve conflicting decisions. The primary outcome will be the identification of staphylococcal enterotoxins in food, and the secondary outcomes will include staphylococcal enterotoxin types and foods involved. To assess the risk of bias in the studies, the tool developed by the Joanna Briggs Institute (JBI) will be used. For data synthesis, a meta-analysis will be performed. However, in case that is not possible, a narrative synthesis of the most relevant results will be carried out. DISCUSSION: This protocol will serve as the basis for a systematic review that aims to relate the results of existing studies on the staphylococcal enterotoxin prevalence and types in food, and the profile of the contaminated foods. The results will broaden the perception of food safety risks, highlight existing literature gaps, contribute to the study of the epidemiological profile and may guide the allocation of health resources for the development of preventive measures related. SYSTEMATIC REVIEW REGISTRATION: PROSPERO registration number: CRD42021258223.

Genomic characterization, antimicrobial resistance profiles, enterotoxin, and biofilm production of methicillin-resistant Staphylococcus aureus isolated from clinical and animal products origins in Eastern Turkey / Caracterização genômica perfis de resistência antimicrobiana, produção de enterotoxinas e biofilme Staphylococcus aureus resistente à meticilina isolado de origem clínica e de produtos de origem animal no leste da Turquia

Staphylococcus aureus is an opportunistic and ubiquitous pathogen found in the skin, nares, and mucosal membranes of mammals. Increasing resistance to antimicrobials including methicillin has become an important public concern. One hundred and eight (108) S. aureus strains isolated from a total of 572 clinical and animal products samples, were investigated for their biofilm capability, methicillin resistance, enterotoxin genes, and genetic diversity. Although only one strain isolated from raw retail was found as a strong biofilm producer, the percentage of antimicrobial resistance pattern was relatively higher. 17.59% of S. aureus strains tested in this study were resistant to cefoxitin and identified as methicillin-resistant S. aureus (MRSA) isolates. mecA and mecC harboring S. aureus strains were detected at a rate of 2.79% and 0.93%, respectively. In addition, staphylococcal enterotoxin genes including Sea, Seb, Sec, and Sed genes were found to be 18.5%, 32.4%, 6.5% and 3.7%, respectively. The phylogenetic relationship among the isolates showed relationship between joint calf and cow milk isolates. Multi locus sequence typing (MLST) revealed three different sequence types (STs) including ST84, ST829, and ST6238. These findings highlight the development and spread of MRSA strains with zoonotic potential in animals and the food chain throughout the world.

Staphylococcus aureus é um patógeno dúctil e ubíquo encontrado na pele, narinas e membranas mucosas de mamíferos. O aumento da resistência aos antimicrobianos, incluindo a meticilina, tornou-se uma importante preocupação pública. Cento e oito (108) cepas de S. aureus isoladas de um total de 572 amostras clínicas e de produtos animais foram investigadas por sua capacidade de biofilme, resistência à meticilina, genes de enterotoxinas e diversidade genética. Embora apenas uma cepa isolada do cru tenha sido encontrada como forte produtora de biofilme, a porcentagem do padrão de resistência antimicrobiana foi relativamente maior. Parte das cepas (17,59%) de S. aureus testadas neste estudo eram resistentes à cefoxitina e identificadas como isolados de MRSA. mecA e mecC abrigando cepas de S. aureus foram detectados a uma taxa de 2,79% e 0,93%, respectivamente. Além disso, verificou-se que os genes da enterotoxina estafilocócica, incluindo os genes Sea, Seb, Sec e Sed, eram 18,5%, 32,4%, 6,5% e 3,7%, respectivamente. A relação filogenética entre os isolados mostrou relação entre os isolados de bezerro e leite de vaca. A tipagem de sequência multiloco (MLST) revelou três tipos de sequência diferentes (STs), incluindo ST84, ST829 e ST6238. Essas descobertas destacam o desenvolvimento e a disseminação de cepas de MRSA com potencial zoonótico em animais e na cadeia alimentar em todo o mundo.

High production of egc2-related staphylococcal enterotoxins caused a food poisoning outbreak.

Staphylococcal enterotoxins (SEs) produced by enterotoxigenic Staphylococcus aureus in food cause staphylococcal food poisoning. We recently reported a foodborne outbreak due to S. aureus harboring new SE/SE-like (SEl) genes (seg, sei, sem, sen, seo, and selu) related to enterotoxin gene cluster (egc) 2 as with other research groups. However, the pathogenicity of SEs production remains unclear. Therefore, we herein investigated egc2-related SEs production from S. aureus isolates and leftover food items during a foodborne outbreak using a sandwich enzyme-linked immunosorbent assay suitable for the quantification of SEs. S. aureus isolates produced markedly high levels of egc2-related SEs, and the leftover food item "Sushi" contained SEs over the toxin dose that causes food poisoning symptoms. A representative isolate was subjected to whole-genome sequencing. The isolate was homologous with previously reported ST45 strains, particularly the unique genomic island νSaß structure mostly consisting of egc2. The present study indicates that egc2-related SEs are food poisoning causative agents based on high SE production levels within an actual foodborne outbreak.

A novel staphylococcal enterotoxin SE02 involved in a staphylococcal food poisoning outbreak that occurred in Tokyo in 2004.

Staphylococcal enterotoxins (SEs) are extracellular proteins, produced mainly by Staphylococcus aureus, which cause staphylococcal food poisoning (SFP) when ingested. Here, a novel SE was identified from two strains, which were identified as the causative microbes of the SFP outbreak that occurred in Tokyo in 2004. Both strains harbored the SEA gene, but its production was lower than that of other SEA-producing SFP isolates. Whole-genome sequencing analysis demonstrated that both strains harbored a SE-like gene besides sea. Phylogenetic analysis revealed that the amino acid sequence deduced from the SE-like gene belonged to the SEB group. Therefore, this gene was presumed to be a novel SE gene and termed "SE02." The stability of SE02 against heating and proteolytic digestions was a little different from that of SEA. SE02 has both superantigenic and emetic bioactivities. Namely, SE02 activated mouse splenocytes and exhibited emetic activity in the common marmoset. SE02 mRNA was highly expressed in both isolates during the exponential phase of cultivation. In addition, SE02 protein was produced at 20 °C and 25 °C, which reflects the actual situation of SFP. SE02 appears to be a novel emetic toxin that was likely the causative toxin in combination with SEA in the SFP outbreak.

Characterization of Oxacillin-Susceptible mecA-Positive Staphylococcus aureus from Food Poisoning Outbreaks and Retail Foods in China.

In this study, we explored the prevalence of oxacillin-susceptible mecA-positive Staphylococcus aureus (OS-MRSA) in staphylococcal food poisoning outbreak isolates and foodborne isolates, and then investigated their molecular characteristics, classical staphylococcal enterotoxins (SEs), and drug resistance. Eight (2.9%) of 275 isolates from food poisoning outbreaks and 7 (3.8%) of 184 isolates from retail foods were identified as OS-MRSA isolates. Among the 15 OS-MRSA isolates, the most frequently detected toxin genes were hld (100%), hla (93.3%), pvl (80.0%), and hlb (46.7%) followed by seg and seq (33.3%, each), hlg (26.7%), seb and hlgv (20.0%, each), sec, seh, sel, sep, and tst (13.3%, each), and sei, sem, sen, and seo (6.7%, each). None of isolates carried other tested virulence genes. The most frequently detected classical SEs were SEB and SEC (26.7%, each), followed by SEA and SEE (20.0%, each), and SED (6.7%). Resistance was most frequently observed in ampicillin, penicillin, and cefoxitin (100%, each), followed by trimethoprim/sulfamethoxazole (93.3%), erythromycin (73.3%), amoxicillin/clavulanic acid (46.7%), tetracyclines (26.7%), and ciprofloxacin (6.7%). All isolates were susceptible to other tested antibiotics. A dominant molecular type belonged to ST398-IVa-t034 (26.7%), followed by ST59-IVa-t437 (20.0%), ST88-III-t14340 and ST1-IVa-t114 (13.3%, each), and ST5-II-t002, ST630-t4549, ST5-II, and ST4495-t10738 (6.7%, each). Our findings indicated that OS-MRSA strains had a low prevalence rate among outbreak strains and foodborne strains, which frequently harbored SCCmec IVa, and carried a variety of toxin genes, and also expressed numerous classical SEs. In addition, all OS-MRSA isolates were susceptible to the majority of antibacterial agents except ß-lactam. Our study is the first to report that OS-MRSA isolates are associated with food poisoning outbreaks worldwide.

Staphylococcal poisoning during a village festival, Medina, Misamis Oriental, Philippines in 2014.

INTRODUCTION: On 18 August 2014, cases of food poisoning in San Vicente Village were reported to the Event-Based Surveillance & Response Unit of the Philippine Department of Health. An investigation was conducted to identify the implicated source, describe the outbreak and evaluate the risk factors. METHODS: A case-control study was conducted. A suspected case was a previously well individual of Medina who attended the village festival and developed abdominal pain and vomiting with or without nausea, diarrhoea and fever from 18 to 19 August. A confirmed case was a suspected case with a rectal swab positive for bacterial culture. Rectal swabs, water and food samples were sent to the national reference laboratories. Food source and consumption interviews and environmental inspections were conducted. RESULTS: Sixty-four cases and 123 unmatched controls were identified. The median incubation period was 1 hour 15 minutes. Five cases (8%) were positive for Staphylococcus aureus, one (2%) for Aeromonas hydrophilia and one (2%) for Shigella boydii. One (14%) water sample was positive for Aeromonas spp. Of the collected food samples, beef steak was positive for Staphylococcus aureus. Risk factors were consumption of Filipino-style beef stew (odds ratio [OR]: 6.62; 95% confidence interval [CI]: 2.90-15.12) and stir-fried noodles (OR: 3.15; 95% CI: 1.52-6.50). Prolonged serving time and improper food storage were noted. DISCUSSION: In this foodborne outbreak, Staphylococcus aureus was the likely causative agent. Meals were contaminated due to improper food handling practices. We recommend that a policy be created to mandate that village-appointed food handlers undergo food safety training.

Genotypes, Enterotoxin Gene Profiles, and Antimicrobial Resistance of Staphylococcus aureus Associated with Foodborne Outbreaks in Hangzhou, China.

Staphylococcal food poisoning is an illness caused by the consumption of food that contains sufficient amounts of one or more enterotoxins. In the present study, a total of 37 S. aureus isolates were recovered from leftover food, swabs from a kitchen environment, and patient feces associated with four foodborne outbreaks that occurred in Hangzhou, southeast China, and were characterized by multilocus sequence typing (MLST), spa typing, pulse-field gel electrophoresis (PFGE), and antimicrobial susceptibility. Classical enterotoxin and enterotoxin-like genes were profiled by PCR analysis. ST6-t304 was the most common clone (40.54%), followed by ST2315-t11687 (32.43%). Six clusters (A to F) were divided based on PFGE patterns, and Clusters A and C were the most common types, constituting 86.49% of all isolates. Moreover, sea was the most frequently identified enterotoxin gene (81.08%), followed by the combination of seg-sei-selm-seln-sleo-selu and sec-sell (each 48.65%). Five isolates also harbored the exotoxin cluster sed-selj-ser. In addition, resistance to penicillin (97.30%), erythromycin (37.85), tetracycline (32.43%), clindamycin, gentamicin, and sulfamethoxazole (each 10.81%) was observed. Our research demonstrated the link between leftover foods and patients by molecular typing and detecting the profiles of enterotoxin or enterotoxin-like genes in human and food isolates. S. aureus maintains an extensive repertoire of enterotoxins and drug resistance genes that could cause potential health threats to consumers.

The occurrence of Staphylococcus aureus, enterotoxigenic and methicillin-resistant strains in Iranian food resources: a systematic review and meta-analysis.

INTRODUCTION: Staphylococcus aureus, in particular its enterotoxigenic strains, is a well-known pathogen in food microbiology as one of the most common causes of food poisoning. This study aimed to investigate the prevalence of S. aureus enterotoxigenic strains in Iranian food resources by using a systematic review and meta-analysis. METHODS: A systematic search was performed using electronic databases from papers that were published by Iranian authors to the end of July 2017. Then, 34 publications which met our inclusion criteria were selected for data extraction and analysis. RESULTS: The average pooled prevalence of S. aureus in food resources was 15.5% (95% CI: 12.7%-18.9%) ranging from 1.1% to 52%. According to the examined publications the estimated average prevalence of enterotoxigenic strains was estimated at 53.7% (95% CI: 41.4%-65.6%) ranging from 13% to 83.3%. The average prevalence of methicillin-resistant S. aureus in contaminated resources was estimated at 15.5% (95% CI: 11.1%-21.4%) ranging from 0% to 32%. The results showed that SEA and SEG were the most prevalent classic and new staphylococcal enterotoxins' (SEs) types. The results of antibiotic resistance pattern showed the highest resistance rates to ß-lactams, tetracycline and erythromycin. CONCLUSIONS: The present analysis showed a significant presence of enterotoxigenic strains of S. aureus and a high rate of staphylococcal enterotoxins among food resources in Iran. However, also the prevalence of MRSA strains is remarkable and requires an improvement of the hygienic conditions to reduce the risk of MRSA dissemination in community and hospitals. Furthermore, the rational use of antibiotics in the environments related to food resources is recommended.

Characterization of A Staphylococcal Food Poisoning Outbreak in A Workplace Canteen during the Post-Earthquake Reconstruction of Central Italy.

In summer 2017, a foodborne outbreak occurred in Central Italy, involving 26 workers employed in the post-earthquake reconstruction. After eating a meal provided by a catering service, they manifested gastrointestinal symptoms; 23 of them were hospitalized. The retrospective cohort study indicated the pasta salad as the most likely vehicle of poisoning. Foods, environmental samples, and food handlers' nasal swabs were collected. Bacillus cereus (Bc) and coagulase-positive staphylococci (CPS) including S. aureus, together with their toxins, were the targets of the analysis. CPS, detected in all the leftovers, exceeded 105 CFU/g in the pasta salad, in which we found Staphylococcal Enterotoxins (SEs) (0.033 ng SEA/g; 0.052 ng SED/g). None of the environmental and human swabs showed contamination. We characterized 23 S. aureus from foods. They all belonged to the human biotype, showed the same toxigenic profile (sea, sed, sej, and ser genes), and had the same Pulsed Field Gel Electrophoresis (PFGE) pattern; none of them harbored mecA or mupA genes. We also detected Bc contamination in the pasta salad but none of the isolates harbored the ces gene for the emetic toxin cereulide. The EU Reference Laboratory for CPS confirmed the case as a strong-evidence outbreak caused by the ingestion of SEs produced by a single strain of S. aureus carried by the same human source. This outbreak was successfully investigated despite the emergency situation in which it occurred.

Prevalence of Enterotoxin Genes in Staphylococcus aureus Isolates from Pork Production.

In this study, 130 Staphylococcus aureus isolates from samples associated with pork production were tested for prevalence of 18 staphylococcal enterotoxin (SE) genes. Approximately 94.6% (123/130) of isolates from different stages of pork production harbored one or more SE genes forming 37 different enterotoxin gene profiles. Seb was present in 60.0% of the S. aureus isolates, the highest among the genes tested. The genes, sed, sej, seo, sep, ser, and seu, were not found. The five classical SE genes (including sea, seb, sec, sed, see) had lower prevalence than the egc gene cluster (seg, sei, sem, sen, seo, or seu). Notably, ∼6.9% (9/130) isolates harbored five SE genes. Classical SE genes were relatively higher in raw meat isolates than swine farm isolates, suggesting that raw meat isolates have a greater potential for classical staphylococcal food poisoning. Incomplete egc clusters were mainly distributed in swine farm isolates, and some of them coexisted with other classical SE genes (seb, sec), showing that swine farms could be potential sources of enterogenic S. aureus of food safety concern. Characterizing the distributions of enterotoxin genes among S. aureus may provide epidemiological information for the benefit of public health and food safety.

From Commensal to Consumer: Staphylococcus aureus Toxins, Diseases, and Detection Methods.

Staphylococcus aureus is a Gram-positive bacterium capable of causing a wide array of infections. Generally a commensal organism, S. aureus encodes several virulence mechanisms that contribute to disease progression. This review highlights toxins as a secreted virulence factor by S. aureus, the diseases that manifest as a result, and the methods used to detect them. In particular, the advantages and limitations of current toxin detection methods are discussed.

Food-Borne Outbreak Investigation and Molecular Typing: High Diversity of Staphylococcus aureus Strains and Importance of Toxin Detection.

Staphylococcus aureus is an important aetiological agent of food intoxications in the European Union as it can cause gastro-enteritis through the production of various staphylococcal enterotoxins (SEs) in foods. Reported enterotoxin dose levels causing food-borne illness are scarce and varying. Three food poisoning outbreaks due to enterotoxin-producing S. aureus strains which occurred in 2013 in Belgium are described. The outbreaks occurred in an elderly home, at a barbecue event and in a kindergarten and involved 28, 18, and six cases, respectively. Various food leftovers contained coagulase positive staphylococci (CPS). Low levels of staphylococcal enterotoxins ranging between 0.015 ng/g and 0.019 ng/g for enterotoxin A (SEA), and corresponding to 0.132 ng/g for SEC were quantified in the food leftovers for two of the reported outbreaks. Molecular typing of human and food isolates using pulsed-field gel electrophoresis (PFGE) and enterotoxin gene typing, confirmed the link between patients and the suspected foodstuffs. This also demonstrated the high diversity of CPS isolates both in the cases and in healthy persons carrying enterotoxin genes encoding emetic SEs for which no detection methods currently exist. For one outbreak, the investigation pointed out to the food handler who transmitted the outbreak strain to the food. Tools to improve staphylococcal food poisoning (SFP) investigations are presented.

Food poisoning outbreak in Tokyo, Japan caused by Staphylococcus argenteus.

Staphylococcus argenteus is a novel species subdivided from Staphylococcus aureus. Whether this species can cause food poisoning outbreaks is unknown. This study aimed to investigate the enterotoxigenic activities of two food poisoning isolates suspected to be S. argenteus (Tokyo13064 and Tokyo13069). The results for phylogenic trees, constructed via whole genome sequencing, demonstrated that both isolates were more similar to a type strain of S. argenteus (MSHR1132) than any S. aureus strain. Moreover, the representative characteristics of S. argenteus were present in both strains, namely both isolates belong to the CC75 lineage and both lack a crtOPQMN operon. Thus, both were determined to be "S. argenteus." The compositions of the two isolates' accessory elements differed from those of MSHR1132. For example, the seb-related Staphylococcus aureus pathogenicity island, SaPIishikawa11, was detected in Tokyo13064 and Tokyo13069 but not in MSHR1132. Both isolates were suggested to belong to distinct lineages that branched off from MSHR1132 lineages in terms of accessory elements. Tokyo13064 and Tokyo13069 expressed high levels of s(arg)eb and produced S(arg)EB protein, indicating that both have the ability to cause food poisoning. Our findings suggest that S. argenteus harboring particular accessory elements can cause staphylococcal diseases such as food poisoning, similarly to S. aureus.

Prevalence and expression of staphylococcal enterotoxin genes in Staphylococcus aureus isolated from food poisoning outbreaks.

Staphylococcus aureus is an important pathogen of foodborne origin. The pathogen produces a variety of toxins that include the staphylococcal enterotoxins (SE). The present study aimed to evaluate the prevalence and expression of 5 SE genes (sea, seb, sec, sed, and see) in S. aureus isolated from outbreaks occurred in the state of Rio Grande do Sul, Brazil. All isolates, with the exception of 2, presented the same or higher transcriptional expression than the reference strains for at least 1 of these genes. The presence of SE genes combined with high levels of transcriptional expression suggests that 1 or more SEs were involved with the staphylococcal food poisoning outbreak analyzed in the present study.