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Master's Dissertation
Document
Author
Full name
Rosa Liliana Solis Castro
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2017
Supervisor
Committee
Cai, Silvana (President)
Ferreira, Rita de Cassia Cafe
Rodrigues, Josias
Sircili, Marcelo Palma
Title in Portuguese
Determinação dos principais patótipos de Escherichia coli isoladas de pacientes com câncer de reto.
Keywords in Portuguese
Escherichia coli
Câncer de reto
ERIC-PCR
Fatores de virulência
Filogrupos
Patótipos
Abstract in Portuguese
No Brasil, os cânceres de cólon e de reto são considerados as neoplasias gastrintestinais mais comumente observadas na população. Nos últimos anos vêm se relatando na literatura nacional e internacional a possível relação da presença de microrganismos com o desenvolvimento de câncer; entretanto, ainda não se observam evidências científicas convincentes dessa interação. Este estudo teve como objetivo determinar a presença e participação dos diferentes patótipos de Escherichia coli em pacientes com e sem câncer de reto. Foram coletadas amostras fecais de pacientes com neoplasia de reto, e de indivíduos sadios sem sinais de câncer (pólipos e/ou tumor), usadas como controle. Uma porção fecal foi semeada em ágar MacConkey isolando-se aletaoriamente quatro colônias de cada amostra. A identificação em nível de espécie, e dos patótipos, assim como dos fatores de virulência das cepas extra-intestinais de E. coli foi realizada por PCR convencional. Para a caracterização molecular das E. coli foi usada a técnica de ERIC-PCR. Os pacientes com neoplasia de reto apresentaram idade média de 63 anos de idade (P < 0,001). A ocorrência de E. coli extra-intestinal (ExPEC; 44,17%) foi menor que as cepas de E. coli diarreiogênicas (DEC; 49,2%). A presença de E. coli enteroagregativa típica (tEAEC) foi observada em 44,1% das amostras fecais de pacientes com câncer de reto e em indivíduos sadios (12,9%) (P = 0,003); entretanto, as E. coli enteropatogênicas atípicas (aEPEC) foram isoladas em ambos os grupos de pacientes (câncer: 37,3%; sadios: 48,4%). O gene afa/dra da adesina Afa/Dr foi observado em maior prevalência nas ExPEC isoladas de pacientes com câncer (P < 0,001). As cepas de E. coli mostraram combinações gênicas que variaram de 2 a 8 genes, observando-se 39 e 24 combinações gênicas nas cepas de pacientes com câncer e sadios, respectivamente. Pelo ERIC-PCR observou-se elevada diversidade genética em todas as cepas. Foi observada a presença dos oito filogrupos de E. coli, sendo o filogrupo B2 (55,2%) o mais predominante. Os filogrupos D e E não agruparam cepas de indivíduos sadios. Os resultados sugerem maiores estudos para determinar o papel das DEC, particularmente das aEPEC, tEAEC e ExPEC de forma individual ou em associação, avaliando-se o provável sinergismo e/ou a co-infecção de diferentes patótipos nesses processos, assim como sua presença no trato intestinal em pacientes assintomáticos com câncer de reto.
Title in English
Determination of the main pathotypes of Escherichia coli in patients with rectal cancer.
Keywords in English
Escherichia coli
ERIC-PCR
Pathotypes
Phylogroups
Rectal cancer
Virulence factors
Abstract in English
In Brazil, colon and rectal cancer are increasing and they are considered the gastrointestinal neoplasia most commonly observed in the population. In recent years, national and international literatures have shown a possible correlationship among the presence of microorganisms with the development of cancer; however, no convincing scientific evidence of this interaction has been observed. This study aimed to determine the presence and participation of different pathotypes of Escherichia coli isolated from patients with or without rectal cancer. Fecal samples were collected from patients with rectal cancer and healthy individuals with no signs of cancer (polyps and/or tumor) used as a control. Feces were plated onto agar MacConkey and four strains were randomly selected from each sample. Conventional PCR was used for identification of E. coli and pathotypes, as well as to detect virulence genes in extra-intestinal strains. The molecular characterization of E. coli was performed by ERIC-PCR. Patients with rectal cancer were mean age of 63 years old (P < 0.001). Diarrheogenic E. coli (DEC, 49.17%) were more prevalent than extra-intestinal E. coli (ExPEC, 44.17%). The presence of tEAEC was observed in 44.1% of the patients with cancer compared to healthy (12.9%) (P = 0.003). Atypical enteropathogenic E. coli (aEPEC) strains were isolated in both patient groups (cancer: 37.3%; healthy: 48.4%). The gene afa/dra for adhesion Afa/Dr was observed in higher prevalence than in ExPEC strains in patients with cancer and healthy subjects (P < 0.001). E. coli strains showed genetic combinations from 2 to 8 genes, showed 39 and 24 genetic combinations in strains from cancer and healthy patients, respectively. All strains showed high genetic diversity by ERIC-PCR. It was observed presence of eight filogroups and B2 filogroup (55.2%) was the most prevalent. Filogroups D and E were absent in strains from healthy. The results suggest further studies to determine the role of DEC, particularly aEPEC, tEAEC, and ExPEC, individually or in combination, and the synergism and co-infection of different pathotypes in these processes, as well as its presence in the intestinal microbiota in asymptomatic patients with rectal cancer.
 
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Release Date
2020-03-12
Publishing Date
2018-03-13
 
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