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In Vitro Assay of Bacterial Adhesion onto Mammalian Epithelial Cells
Published on: May 16, 2011
Pseudobutyrivibrio xylanivorans adhesion to epithelial cells
Diego Javier Grilli1, Maria Eugenia Mansilla2, María Cecilia Giménez3
1Facultad de Ciencias Veterinarias y Ambientales, Universidad Juan Agustín Maza, Av. Acceso Este Lateral Sur 2245, CP 5519, Mendoza, Argentina; Instituto de Histología y Embriología de Mendoza, Universidad Nacional de Cuyo, Casilla de Correo 56, CP 5500, Mendoza, Argentina; Área de Microbiología, Departamento de Patología, Facultad de Ciencias Médicas, Universidad Nacional de Cuyo, Casilla de Correo 56, CP 5500, Mendoza, Argentina.
Abstract:
The ruminal bacteria Pseudobutyrivibrio xylanivorans strain 2 (P. xylanivorans 2), that mediates the digestion of plant fiber, is considered an attractive candidate for probiotics. Adherence to the epithelium of the digestive tract of the host is one of the major requirements for probiotics. In this study, we assessed the adhesion of P. xylanivorans 2 to SW480 cells and characterized this process utilizing multiple microscopy approaches. Our results indicate that a multiplicity of infection of 200 CFU/cell allows the highest bacteria to cell binding ratio, with a lower percentage of auto-agglutination events. The comparison of the adherence capacity subjected heat-shock treatment (100 °C, 1 min), which produces the denaturalization of proteins at the bacterial surface, as opposed untreated P. xylanivorans, suggested that this bacteria may attach to SW480 cells utilizing a proteinaceous structure. Confocal microscopy analyses indicate that P. xylanivorans 2 attachment induces the formation of F-actin-enriched areas on the surface of SW480 cells. Transmission electron microscopy (TEM) revealed the formation of a structure similar to a pedestal in the area of the epithelial cell surface, where the bacterium rests. Finally, a casual finding of TEM analysis of transverse and longitudinal thin-sections of P. xylanivorans 2, revealed irregular intra-cytoplasmic structures compatibles with the so-called bacterial microcompartments. This is the first ultrastructural description of bacterial microcompartments-like structures in the genus Pseudobutyrivibrio.
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