Charlène Roussel1,2, Charlotte Cordonnier1,3, Wessam Galia1,4,5, Olivier Le Goff1, Jonathan Thévenot1,3, Sandrine Chalancon1, Monique Alric1, Delphine Thevenot-Sergentet4,6, Francoise Leriche5, Tom Van de Wiele2, Valérie Livrelli3,7, Stéphanie Blanquet-Diot1. 1. EA 4678 CIDAM, Conception Ingénierie et Développement de l'Aliment et du Médicament, Université d'Auvergne, Clermont-Ferrand, France. 2. CMet, Center for Microbial Ecology and Technology, Ghent University, Ghent, Belgium. 3. M2iSH, Microbes, Intestin, Inflammation et Susceptibilité de l'Hôte UMR INSERM/Université d'Auvergne, Université d'Auvergne, Clermont-Ferrand, France. 4. UMR 5557 Ecologie Microbienne, Research Group on Bacterial Opportunistic Pathogens and Environment, CNRS, VetAgro Sup and Université de Lyon, Lyon, France. 5. Unité CALYTISS, VetAgro Sup, Lempdes, France. 6. Laboratoire d'Etude des Microorganismes Alimentaires Pathogènes, French National Reference Laboratory for Escherichia coli including Shiga Toxin-Producing E. coli, VetAgro Sup, Université de Lyon, Marcy l'Etoile, France. 7. Service de Bactériologie, CHU de Clermont-Ferrand, Clermont-Ferrand, France.
Abstract
BACKGROUND: Enterohemorrhagic Escherichia coli (EHEC) are major foodborne pathogens that constitute a serious public health threat, mainly in young children. Shiga toxins (Stx) are the main virulence determinants of EHEC pathogenesis but adhesins like intimin (eae) and Long polar fimbriae (Lpf) also contribute to infection. The TNO GastroIntestinal Model (TIM) was used for a comparative study of EHEC O157:H7 survival and virulence under adult and child digestive conditions. METHODS: Survival kinetics in the in vitro digestive tract were determined by plating while bacterial viability was assessed by flow cytometry analysis. Expression of stx, eae, and lpf genes was followed by reverse transcriptase-quantitative PCR (RT-qPCR) and Stx production was measured by ELISA (enzyme-linked immunosorbent assay). RESULTS: Upon gastrointestinal passage, a higher amount of viable cells was found in the simulated ileal effluents of children compared to that of adults (with 34 and 6% of viable cells, respectively). Expression levels of virulence genes were up to 125-fold higher in children. Stx was detected only in child ileal effluents. CONCLUSION: Differences in digestive physicochemical parameters may partially explain why children are more susceptible to EHEC infection than adults. Such data are essential for a full understanding of EHEC pathogenesis and would help in designing novel therapeutic approaches.
BACKGROUND: Enterohemorrhagic Escherichia coli (EHEC) are major foodborne pathogens that constitute a serious public health threat, mainly in young children. Shiga toxins (Stx) are the main virulence determinants of EHEC pathogenesis but adhesins like intimin (eae) and Long polar fimbriae (Lpf) also contribute to infection. The TNO GastroIntestinal Model (TIM) was used for a comparative study of EHEC O157:H7 survival and virulence under adult and child digestive conditions. METHODS: Survival kinetics in the in vitro digestive tract were determined by plating while bacterial viability was assessed by flow cytometry analysis. Expression of stx, eae, and lpf genes was followed by reverse transcriptase-quantitative PCR (RT-qPCR) and Stx production was measured by ELISA (enzyme-linked immunosorbent assay). RESULTS: Upon gastrointestinal passage, a higher amount of viable cells was found in the simulated ileal effluents of children compared to that of adults (with 34 and 6% of viable cells, respectively). Expression levels of virulence genes were up to 125-fold higher in children. Stx was detected only in child ileal effluents. CONCLUSION: Differences in digestive physicochemical parameters may partially explain why children are more susceptible to EHEC infection than adults. Such data are essential for a full understanding of EHEC pathogenesis and would help in designing novel therapeutic approaches.
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