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Master's Dissertation
DOI
https://doi.org/10.11606/D.87.2014.tde-06122014-083305
Document
Author
Full name
Cristiane Moda Mota
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2014
Supervisor
Committee
Franzolin, Marcia Regina (President)
Martins, Elizabeth Angelica Leme
Neves, Carla Taddei de Castro
Title in Portuguese
Influência de diferentes condições de cultivo na formação de biofilme por Escherichia coli enteropatogênica atípica (EPECa) e pesquisa de genes relacionados.
Keywords in Portuguese
Escherichia coli
Biofilme
Diarreia
EPEC atípica
Fatores de virulência
Película
Abstract in Portuguese
EPECa tem sido identificada como o principal agente de diarreia aguda em crianças de países em desenvolvimento. Biofilmes são estruturas bacterianas envoltas por uma matriz de exopolissacarídeos. O objetivo deste estudo foi verificar a influência de diferentes condições de cultivo na formação de biofilme por EPECa isoladas de crianças com diarreia e pesquisar a presença de genes relacionados com a formação de biofilme. As sequências genéticas dos genes csgA, crl, fimH e bcsA foram pesquisadas através da PCR e verificadas em 6 (26%), 23 (100%), 22 (95,6%) e 23 (100%) das amostras respectivamente. A formação de biofilme em placas de MTP por EPECa foi melhor evidenciada em meio LB sem sal a 26 °C e em caldo E. coli a 37 °C, tanto em número de amostras quanto em intensidade da formação de biofilme. As microscopias confocal e de varredura propiciaram uma análise qualitativa de microcolônias e pilares, além de EPS respectivamente. As condições de cultivo devem ser usadas com precaução para realmente aferir a capacidade de formação de biofilme por amostras de EPECa.
Title in English
Influence of different culture conditions on biofilm formation by atypical enteropathogenic Escherichia coli (aEPEC) and search of related genes.
Keywords in English
Escherichia coli
Atypical EPEC
Biofilm
Diarrhea
Pellicle
Virulence factors
Abstract in English
aEPEC has been identified as the main agent of acute diarrhea in children in developing countries. Biofilms are bacterial structures which are surrounded by a matrix of exopolysaccharides. The aim of this study was investigate the influence of different culture conditions on biofilm formation by 23 aEPEC strains isolated from children with diarrhea and search for the presence of genes related to biofilm formation. The genetic sequences of the csgA, crl, fimH and bcsA genes were screened by PCR and were found in 6 (26%), 23 (100%) 22 (95.6%) and 23 (100%) of the strains respectively. Biofilm formation in MTP plates for aEPEC strains was better evidenced in LB medium without NaCl at 26 ° C and in E. coli broth at 37 ° C, both in number and intensity of biofilm formation. The confocal and scanning electron microscopy provided a qualitative analysis of structures such as microcolonies and pillars, and respectively EPS. The growing conditions should be used with caution to measure the real ability of biofilm formation by strains of aEPEC.
 
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Publishing Date
2014-12-10
 
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