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Master's Dissertation
DOI
https://doi.org/10.11606/D.10.2012.tde-05062013-160623
Document
Author
Full name
Gabriele Bin Alves Pereira
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2012
Supervisor
Committee
Ferreira, Antônio José Piantino (President)
Knöbl, Terezinha
Mettifogo, Elena
Title in Portuguese
Imunodetecção de antígenos de Escherichia coli em infecção experimental em aves
Keywords in Portuguese
Escherichia coli
Western blot
Eletroforese unidimensional
Proteínas
Abstract in Portuguese
A colibacilose é uma enfermidade importante na avicultura por causar prejuízos econômicos e danos à carcaça. Escherichia coli é um micro-organismo oportunista e a complexidade da infecção se deve a diversidade de sorotipos, variados fatores de virulência e baixa capacidade de estimular a imunidade cruzada. Assim, a compreensão dos mecanismos de virulência que ocorrem no hospedeiro, pode resultar no desenvolvimento de métodos diagnósticos e profiláticos. Neste estudo, 85 pintinhos foram divididos em quatro grupos, sendo três infectados com diferentes sorogrupos de Escherichia coli (O2, O78, O119) e um vacinado com Poulvac® E.coli (Pfizer®). As lesões sugestivas de colibacilose foram avaliadas e qualificadas aos 35 dias de idade, e em seguida realizada a colheita de sangue para a obtenção do soro. Foram obtidas proteínas in vitro dos sorogrupos de E. coli utilizados na infecção experimental e da vacina (mutante de Escherichia coli O78). Estas foram separadas por eletroforese unidimensional em gel de acrilamida utilizando dodecil sulfato de sódio (SDS) e foram submetidas a reação de western blot com a mistura de soros de cada grupo estudado e também com o pool de soros de aves livres de patógenos específicos (SPF), usadas como controle negativo. O perfil eletroforético revelou a presença de várias proteínas dos diferentes sorogrupos de E. coli quando cultivadas no meio casaminoácido e levedura. Houve diferenças entre os sorotipos, mas também proteínas comuns. Foi possível estabelecer uma relação entre virulência, quantidades de proteínas específicas e taxa de reisolamento.
Title in English
Imunodetection of Escherichia coli antigens in experimental infection in broiler flocks
Keywords in English
Escherichia coli
Western blot
Proteins
Unidimensional electrophoresis
Abstract in English
The colibacillosis is a serious disease in poultry industry to cause economic losses and damage to carcass. Escherichia coli is an opportunistic microorganism and the complexity of infection is due to the diversity of serotypes, several virulence factors and low ability to stimulate cross-immunity. Thus understanding of virulence mechanisms that occur in the host, may result in development of diagnostic and prophylactic methods. In this study, 85 chicks were divided into four groups being infected with three different serogroups of E. coli (O2, O78, O119) and vaccinated with Poulvac E. coli ® (Pfizer ®). The lesions suggestive of colibacillosis were evaluated at 35 days of age, and then the blood sample are collected to obtain the serum. Proteins were obtained in vitro from serogroups of E. coli used in experimental infection and vaccine (mutant of Escherichia coli O78). These were separated by one-dimensional electrophoresis on acrylamide gels using sodium dodecyl sulfate (SDS) and then reacted by western blot with the pool of sera from each group and also with the pool of sera from specific pathogen free poultry used as negative control. The electrophoretic pattern showed the presence of many proteins of different serogroups of E. coli when grown in the broth casaminoacid and yeast. There were differences among serotypes, but also common proteins. It was possible to establish a relationship between virulence and reisolation rate.
 
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Publishing Date
2013-09-25
 
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