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Doctoral Thesis
DOI
https://doi.org/10.11606/T.42.2008.tde-22092008-140217
Document
Author
Full name
Marilucia dos Santos Ludovico
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2008
Supervisor
Committee
Yano, Tomomasa (President)
Carbonell, Gleize Villela
Castro, Antonio Fernando Pestana de
Knöbl, Terezinha
Moreno, Andrea Micke
Title in Portuguese
Purificação e caracterização de uma toxina enterotóxica, citotóxina e letal produzida por amostras de Plesiomonas shigelloides isoladas de água de rio.
Keywords in Portuguese
Plesiomomas shigelloides
Atividade letal
Cardiotoxina
Citotoxinas
Enterotoxinas
Purificação
Abstract in Portuguese
Amostras de Plesiomonas shigelloides, isoladas de água de rio, produziram uma citotoxina denominada LCF (Fator Citotóxico Letal), ativa em células Vero, CHO, CaCo-2 e HT-29, causando vacuolizações intracelulares e retração dos núcleos, levando á morte por apoptose. Esta citotoxina foi purificada e seu peso molecular estimado em 48 kDA. É uma citotoxina termolábil e foi parcialmente neutralizada por antisoro anti-LT-1 (enterotoxina termolábil tipo 1 de Escherichia coli) e pelo antisoro anti-Aerolisina de Aeromonas. O antisoro anti-CT (enterotoxina colérica) não neutralizou sua atividade. Em teste "Western blot" para anti-LT-1, não ocorreu reação sorológica. O LCF induziu acúmulo de fluido em intestino de camundongos recém-nascidos e alças intestinais de coelhos. O LCF foi letal para camundongos e ratos, levando-os a morte cardíaca súbita quando injetado por via intravenosa. Considerando os resultados obtidos, neste estudo, as amostras de P. shigelloides isolados de água de rio, mostraram grande potencial como patógeno para o homem e animal.
Title in English
Purification and caracterization of an enterotoxic, cytotoxic and lethal toxin produced by Plesiomonas shigelloides isolated from river water.
Keywords in English
Plesiomomas shigelloides
Cardiotoxin
Citotoxin
Enterotoxin
Lethal activity
Purification
Abstract in English
Plesiomonas shigelloides is a ubiquous microorganism recognized as putative human and animal enteropathogen. Production of toxins has been related to their role in pathogenicity. At previous studies we detected a cytotoxin, named LCF (Lethal Citotoxic Factor) active on a variety of cells, causing intensive intracellular vacuolation and nuclear condensation, leading to death. The toxin purification revealed a heat-labile protein of about 48 kDa by electrophoresis analysis in poliacrylamide gel (SDS-PAGE). The cytotoxic activity was partially neutralized by anti-LT-1 (type 1 termolabile enterotoxin of Escherichia coli) and by anti-aerolisin (cytotoxic enterotoxin of Aeromonas). Western blot analysis to anti-LT-1 showed no serological reaction. Fluid accumulation occurred when toxin was applied to the intestine of newborn mice and to rabbit intestinal loop assays. LCF was lethal to mice and rats, leading them to cardiac death when injected intravenously. Considering the results, these isolates of P. shigelloides have shown great potential as pathogens.
 
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Publishing Date
2008-09-24
 
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