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Investigation on physico-chemical processes during adhesion and biofilm development of Xylella fastidiosa

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Author(s):
Gabriela Simone Lorite
Total Authors: 1
Document type: Doctoral Thesis
Press: Campinas, SP.
Institution: Universidade Estadual de Campinas (UNICAMP). Instituto de Física Gleb Wataghin
Defense date:
Examining board members:
Mônica Alonso Cotta; Angela Maria Moraes; Fábio de Lima Leite; Ricardo Meurer Papaléo; Mário Noboru Tamashiro
Advisor: Mônica Alonso Cotta
Abstract

In this work, we report an investigation on relevant physicochemical processes of bacterial surface adhesion and biofilm development for the phytopathogen Xylella fastidiosa (Xf) on different surfaces. This study aims to corroborate or supplement different aspects of the bacterial biofilms models currently under discussion and, in a technological point of view, provides experimental input for an eventual control of Xf biofilm formation. For this purpose, we have used an approach not commonly found in biology ¿ and more similar to materials science ¿ in order to isolate and quantify the relevance of the parameters that contribute to the formation of a biofilm on different surfaces, as well as trying to make these surfaces closer to the plant xylem. Our results show a similar development (shape, size and quantity) for Xf biofilms grown on Si and glass surfaces, and an improved development of biofilms grown on Si than on ethyl cellulose, EC, and cellulose acetate, AC. Under a phenomenological point of view, biofilms present different structures, rate of development and trends in gene expression between surfaces with and without the presence of derivatized cellulose. In order to characterize the surfaces, we considered the effect of culture medium on their properties. In Si and glass surfaces, we observe the formation of a conditioning film due to adsorption of the constituents of the culture medium. The degree of hydrophobicity of glass and Si surfaces decreases significantly after contact with this medium. On the other hand, cellulose derivatized surfaces present lower (EC) or no (AC) modifications of this property. In addition, we observed an increase in the surface potential (SP) for Si and EC and also a SP decrease for AC. These evidences suggest a correlation between higher SP values and lower degree of hydrophobicity with the presence of biofilms of Xf in larger numbers and size. Furthermore, force spectroscopy measurements using a functionalized tip with the adhesion protein XadA1 also show a different behavior on the AC surface. These results reveal the importance of electrostatic interaction in the initial bacterial adhesion for the surfaces studied here. Finally, the presence of extracellular material around the Xf cells and biofilms on glass and Si surfaces was observed indicating the presence of a protective exopolymeric matrix. Infrared spectroscopy shows the presence of polysaccharides - a constituent of the polymeric matrix - from the very initial stages of biofilm formation, indicating a possible contribution of this matrix for the adhesion process which precedes biofilm development (AU)

FAPESP's process: 06/06397-6 - Structural and functional characteristics of biofilms and biopolymers
Grantee:Gabriela Simone Lorite
Support Opportunities: Scholarships in Brazil - Doctorate