On October 1, 2014, children and staff members at a Swiss boarding school consumed Tomme, a soft cheese produced from raw cow milk. Within the following 7h, all 14 persons who ingested the cheese fell ill, including 10 children and 4 staff members. Symptoms included abdominal pain and violent vomiting, followed by severe diarrhea and fever. We aim to present this food poisoning outbreak and characterize the causative agent. The duration of the incubation period was dependent of the age of the patient: 2.5h in children under 10 yr of age, 3.5h in older children and teenagers, and 7h in adults. The soft cheese exhibited low levels of staphylococcal enterotoxin (SE) A (>6ng of SEA/g of cheese) and high levels of staphylococcal enterotoxin D (>200ng of SED/g of cheese). Counts of 10(7) cfu of coagulase-positive staphylococci per gram of cheese were detected, with 3 different Staphylococcus aureus strains being present at levels >10(6) cfu/g. The 3 strains were characterized using spa typing and a DNA microarray. An enterotoxin-producing strain exhibiting sea and sed was identified as the source of the outbreak. The strain was assigned to spa type tbl 3555 and clonal complex 8, and it exhibited genetic criteria consistent with the characteristics of a genotype B strain. This genotype comprises bovine Staph. aureus strains exclusively associated with very high within-herd prevalence of mastitis and has been described as a major contaminant in Swiss raw milk cheese. It is therefore highly likely that the raw milk used for Tomme production was heavily contaminated with Staph. aureus and that levels further increased due to growth of the organism and physical concentration effects during the cheese-making process. Only a few staphylococcal food poisoning outbreaks involving raw milk products have been described. Still, in view of this outbreak and the possible occurrence of other foodborne pathogens in bovine milk, consumption of raw milk and soft cheese produced from raw milk constitutes a health risk, particularly when young children or other members of sensitive populations are involved.

Staphylococcal food poisoning represents the most prevalent foodborne intoxication worldwide. It is caused by oral intake of enterotoxins preformed by Staphylococcus aureus in food. The relevance of newly described enterotoxins in outbreaks of staphylococcal food poisoning is controversially discussed. Although the staphylococcal enterotoxins SEG, SEI, SEM, SEN, and SEO elicit emesis in a monkey feeding assay, there has been no conclusive proof of their emetic activity in humans. In this study, we provide further evidence suggesting that one of these enterotoxins or a combination of SEG, SEI, SEM, SEN, and SEO cause staphylococcal food poisoning. We investigated two outbreaks registered with the Swiss Federal Office of Public Health, in which only Staphylococcus aureus strains harboring the egc cluster, including seg, sei, sem, sen, and seo linked to typical signs of staphylococcal food poisoning were isolated. The outbreaks were caused by consumption of raw goat cheese and semi-hard goat cheese, and were linked to strains assigned to CC45 (agr type I) and CC9 (agr type II), respectively. These outbreaks provide further evidence that newly-described staphylococcal enterotoxins are likely to cause staphylococcal food poisoning in humans.

Identification of reference genes that can be used to normalize qPCR mRNA expression levels during growth of Staphylococcus aureus in food-related osmotic or la

Dairy goat and sheep farms suffer severe economic losses due to intramammary infections, with S. aureus representing the main cause of clinical mastitis in small ruminants. In addition, S. aureus contamination of goat and sheep milk may cause staphylococcal food poisoning, as many traditional caprine and ovine milk products are not subjected to pasteurization. Data on virulence and antimicrobial resistance genes, as well as on the clonality of S. aureus detected in goat and sheep milk is scarce. Therefore, it was the aim of this study to determine i) spa types and clonal complexes and ii) virulence and resistance gene profiles of S. aureus isolated from goat and sheep milk. A total of 162 milk samples from sheep and goats presenting signs of an intramammary infection and 104 bulk milk samples were collected. While low prevalence rates of S. aureus were detected on single animal level, 46% of the bulk tank milk samples from small ruminants were positive for S. aureus. All isolates were spa typed and clonal complexes and virulence and resistance gene patterns were determined using a DNA microarray. Data from 49 S. aureus isolates was included in the statistical analysis and the construction of a SplitsTree. The analyzed isolates could be assigned to eleven clonal complexes, with the large majority of goat and sheep isolates being assigned to CC130 and CC133. The findings of this study suggest that S. aureus shows pronounced adaptation to small ruminants in general, but not to sheep or goats in particular. Although some common characteristics among S. aureus from caprine, ovine, and bovine milk samples were observed, S. aureus from small ruminants seem to form a distinct population. As 67% of the detected S. aureus strains exhibited at least one enterotoxin gene, many caprine or ovine raw milk products may be contaminated with low levels of enterotoxigenic S. aureus, stressing the importance of strict maintenance of the cold chain.