A cluster of 34 human cases of listeriosis was traced to consumption of contaminated quargel cheese, a sour milk specialty sold in Austria, Germany and Czech Republic. Here, we try to assess how many portions were consumed by the Austrian population at a certain contamination level (CL). In total, 1623 cheese lots were produced during the outbreak period resulting in >3 million portions of cheese delivered to the market. From 650 sets of quality control data provided by the food business operator, we reconstructed the contamination scenario over time and identified 84 lots that were found to be positive. With regard to another sixteen lots, a CL was found ranging from one to 3,84 log10 CFU L. monocytogenes/g, measured in product stored between one to 23 days after production. However the number of storage days at home before consumption is unknown. To resolve this issue, we modelled the theoretical CL of the product if consumed either 20, 30, 40 or 50 days post production. We found that 10 lots (approx. 27,350 portions) would have been contaminated at CLs higher than 3 log10 CFU L. monocytogenes/g if all cheese had been consumed after 20 days of storage. This number shifts to 20 lots (approx. 54,700 portions) after 30 days of storage. If all cheese had been consumed at the end of shelf life (50 days of storage), theoretically 242,5 lots would have exceeded a CL of 6 log10 CFU L. monocytogenes/g. We concluded that the extended shelf life given to the product was a driver of the outbreak scenario. It is stunning to note that so few cases were reported in spite of consumers' massive exposure to L. monocytogenes. We hypothesized that a low pathogenicity of both quargel outbreak clones (QOC1 and QOC2) could have contributed to this discrepancy. Our hypothesis was falsified since both strains QOC1 and QOC2 are fully virulent in an oral infection mouse model, showing even higher pathogenicity than the reference strain EGDe.

• MeSH
• Disease Outbreaks * MeSH
• Food Contamination analysis   MeSH
• Middle Aged MeSH
• Humans MeSH
• Listeria monocytogenes growth & development   isolation & purification   MeSH
• Listeriosis epidemiology   microbiology   mortality   MeSH
• Milk microbiology   MeSH
• Mice, Inbred BALB C MeSH
• Mice MeSH
• Food Microbiology MeSH
• Retrospective Studies MeSH
• Quality Control MeSH
• Aged, 80 and over MeSH
• Aged MeSH
• Cheese microbiology   MeSH
• Virulence MeSH
• Animals MeSH
• Check Tag
• Middle Aged MeSH
• Humans MeSH
• Male MeSH
• Mice MeSH
• Aged, 80 and over MeSH
• Aged MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Geographicals
• Czech Republic MeSH
• Germany MeSH
• Austria MeSH

A large listeriosis outbreak occurred in Austria, Germany and the Czech Republic in 2009 and 2010. The outbreak was traced back to a traditional Austrian curd cheese called "Quargel" which was contaminated with two distinct serovar 1/2a Listeria monocytogenes strains (QOC1 and QOC2). In this study we sequenced and analysed the genomes of both outbreak strains in order to investigate the extent of genetic diversity between the two strains belonging to MLST sequence types 398 (QOC2) and 403 (QOC1). Both genomes are highly similar, but also display distinct properties: The QOC1 genome is approximately 74 kbp larger than the QOC2 genome. In addition, the strains harbour 93 (QOC1) and 45 (QOC2) genes encoding strain-specific proteins. A 21 kbp region showing highest similarity to plasmid pLMIV encoding three putative internalins is integrated in the QOC1 genome. In contrast to QOC1, strain QOC2 harbours a vip homologue, which encodes a LPXTG surface protein involved in cell invasion. In accordance, in vitro virulence assays revealed distinct differences in invasion efficiency and intracellular proliferation within different cell types. The higher virulence potential of QOC1 in non-phagocytic cells may be explained by the presence of additional internalins in the pLMIV-like region, whereas the higher invasion capability of QOC2 into phagocytic cells may be due to the presence of a vip homologue. In addition, both strains show differences in stress-related gene content. Strain QOC1 encodes a so-called stress survival islet 1, whereas strain QOC2 harbours a homologue of the uncharacterized LMOf2365_0481 gene. Consistently, QOC1 shows higher resistance to acidic, alkaline and gastric stress. In conclusion, our results show that strain QOC1 and QOC2 are distinct and did not recently evolve from a common ancestor.

• MeSH
• Cell Line MeSH
• History, 21st Century MeSH
• Species Specificity MeSH
• Disease Outbreaks history   MeSH
• Phylogeny MeSH
• Genetic Variation * MeSH
• Genome, Bacterial genetics   MeSH
• Humans MeSH
• Listeria monocytogenes classification   genetics   pathogenicity   MeSH
• Listeriosis epidemiology   MeSH
• Models, Genetic MeSH
• Molecular Sequence Data MeSH
• Likelihood Functions MeSH
• Base Sequence MeSH
• Sequence Analysis, DNA MeSH
• Virulence MeSH
• Check Tag
• History, 21st Century MeSH
• Humans MeSH
• Publication type
• Journal Article MeSH
• Historical Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Geographicals
• Czech Republic MeSH
• Germany MeSH
• Austria MeSH

For the first time it has been possible to determine the contamination level of Listeria monocytogenes in the very cheese lots of acid curd cheese that caused a multinational outbreak between 2009/2010. The listeriosis outbreak accounted for 34 clinical cases and eight deaths. The cheese, which was distributed in Austria, Germany, the Czech Republic, Poland and Slovakia, was recalled on the 23rd January 2010. All recalled lots were immediately investigated after call back from the retail market. The company manufactured two different cheese types, (i) red smear ripened--and (ii) mold coated/white veined--acid curd cheese. Depending on the lot production dates, cheese samples (n=1045) were analyzed at three different time points: (i) beginning to mid shelf-life (lot nos. 15-18; production period 5.1.2010-13.1.2010); (ii) end of shelf-life (lot nos. 9-18; production period 21.12.2009-13.1.2010) and, (iii) ≤46days after the expiry date (lot nos. 1-18; production period 1.12.2009-13.1.2010). Qualitative and quantitative examinations of cheese samples were performed according to ISO 11290-1&2. Examination of the samples, according to ISO 11290-1, resulted in 16 L. monocytogenes positive (red smear type) and two negative lots (mold coated type). These results were confirmed by a combined enrichment/real-time PCR method. The contamination values obtained by quantitative ISO 11290-2 varied from ≤log 2 cell forming units (CFU)/g to log 8.1CFU/g. Three out of sixteen L. monocytogenes positive lots revealed a contamination level of ≤log 2CFU/g at the beginning of their shelf-life when stored at 4°C. Nevertheless, by increasing the storage life and/or the storage temperature (15, 22°C) the contamination level could be raised to between log 5 and log 6CFU/g. Our data indicate that 81.3% (13/16) of the recalled red smear quargel cheese lots were highly contaminated with L. monocytogenes. All this implies that the main contamination of the quargel cheese took place during the red smear process and that quargel cheese can easily support growth of L. monocytogenes.

• MeSH
• Disease Outbreaks MeSH
• Food Contamination analysis   MeSH
• Listeria monocytogenes growth & development   isolation & purification   MeSH
• Listeriosis MeSH
• Foodborne Diseases MeSH
• Food Microbiology MeSH
• Product Recalls and Withdrawals * MeSH
• Cheese microbiology   poisoning   MeSH
• Temperature MeSH
• Publication type
• Journal Article MeSH
• Geographicals
• Czech Republic MeSH
• Germany MeSH
• Poland MeSH
• Austria MeSH
• Slovakia MeSH