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Caractérisation in situ du développement d'un biofilm par suivi de microbilles à l'aide d'une méthode de corrélation d'images numériques

Abstract : The control of biofilm formation constitutes an important challenge in many industrial and biomedical applications. In this context, BioFilm Control is a pioneer thanks to its test named BioFilm Ring Test. Based on the immobilisation of magnetic microbeads by adherent cells, the assay allows to detect the presence of biofilm at a given time. The aim of this phD project is to translate the BioFilm Ring Test® into a dynamic, non-destructive and microscopic examination of the biofilm state. Whithin the biofilm, the matrix provides a strong cohesion between cells and therefore increases their resistance against chemical or mechanical stress in comparison to their planktonic counterparts. The mechanical properties of the matrix are therefore an indicator of the local state of the biofilm. The search for these parameters makes it possible to predict and control the formation, accumulation and spread of bacteria that propagate infections and/or biofouling. Nevertheless, the determination of the mechanical properties of biofilms requires precautions and the use of an homogenized vocabulary and methods that are unified within the community. To this end, a first part of this thesis work consists in proposing a guide of good practices for the mechanical characterization of biofilm material. In the second part of this work, a methodology for tracking of micrometric particles within a living material is developed. The use of full field measurement method such as digital image correlation makes it possible to trace the kinematics of the motion of each particle, which is a probe of the local environment. This method is then applied to the study of the biofilm formation, by non-contact measurement. The originality of this work is based on the characterization of the change in the microbeads movement during the biofilm formation steps. The aim is to discriminate bead behaviours that reveal the genesis of a biofilm. By taking advantage of the observation of the movement of inert microbeads embedded into the bacterial environment, we detect changes of type of trajectories which seem to be correlated to the activity of sessiles bacteria, adhesion or formation of extracellular material. The results show that the various stages of the biofilm formation are characterized by a non-destructive test. Especially, It allows to appreciate the efficiency of an antibiotic. In the last part, research still in a development phase is presented. It concerns the behaviour of biofilm under mechanical solicitation. This involves observing the biofilm in the far field and following the displacement or deformation of a pattern consisting of an agglomerate of microbeads immersed in a magnetic field. This initial work can be used as a draft for future work to quantitatively characterize the biofilm material.
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Heloïse Boudarel. Caractérisation in situ du développement d'un biofilm par suivi de microbilles à l'aide d'une méthode de corrélation d'images numériques. Matériaux. Université Clermont Auvergne, 2018. Français. ⟨NNT : 2018CLFAC061⟩. ⟨tel-02094489⟩

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