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Colonisation de la viande par Escherichia coli O157∶H7 : caractérisation moléculaire, cellulaire et tissulaire des interactions

Abstract : Escherichia coli O157:H7 is the most prevalent serotype involved in foodborne infection by enterohemorrhagic E. coli (EHEC). It is associated with life-threatening hemorrhagic colitis and the hemolyticuremic syndrome (HUS), which essentially affect young children. The major food vector of EHEC contamination is ground beef. The primary bacterial contamination occurs during the slaughter, essentially at dehiding stage where bacteria can be transferred from hides to carcasses. The connective tissue surrounding the muscle, highly similar to extracellular matrix (ECM) could potentially be a support for bacterial adhesion. When investigating the adhesion and colonization to the main muscle fibrous ECM proteins, the great influence of growth conditions on subsequent bacterial attachment was shown. Maximal adhesion to ECM proteins occurred at 25°C and pH 7, especially to collagens I and III. In EHEC, various surface-exposed protein determinants can be expressed and potentially involved in ECM adhesion. Investigating the autoaggregation, bacterial adhesion and biofilm formation, the involvement of Antigen 43 (Ag43), an autotransporter protein, was demonstrated in E. coli O157:H7 EDL933. Then, the attachment of E. coli O157:H7 to the meat was determined on two different model muscles, with different contractile and metabolic characteristic (Soleus oxidative, slow and EDL glycolytic, fast), previously characterized by UV microspectroscopy coupled to synchrotron radiation fluorescence. The different of muscle fiber types and the effect of a prolonged anoxia simulating maturing meat were discriminated by their spectral responses after excitation at 275 nm. It clearly appeared that bacteria displayed differential tropism as function of the muscle types, higher for the Soleus than the EDL muscles. While E. coli O157:H7 adhered similarly to the different types of muscle fibers, bacterial adherence essentially occurred at the ECM, pinpointing the key role of connective tissue for E. coli O157:H7 adhesion to meat. This first comprehensive investigation of bacterial adhesion to skeletal muscles at molecular, cellular and tissue levels provides new insight in the physiology of the colonization of meat by EHEC and constitutes a prerequisite for the development of innovative practices and strategies to minimize the risk of meat contamination.
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Caroline Chagnot. Colonisation de la viande par Escherichia coli O157∶H7 : caractérisation moléculaire, cellulaire et tissulaire des interactions. Sciences agricoles. Université Blaise Pascal - Clermont-Ferrand II, 2014. Français. ⟨NNT : 2014CLF22448⟩. ⟨tel-01673786v2⟩



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