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Lactobacillus plantarum isolated from kefir protects vero cells from cytotoxicity by type-II shiga toxin from Escherichia coli O157:H7

Published online by Cambridge University Press:  27 November 2012

Emiliano Kakisu ,
Analía G. Abraham ,
Carla Tironi Farinati ,
Cristina Ibarra  and
Graciela L. De Antoni
Emiliano Kakisu*
Affiliation:
Centro de Investigación y Desarrollo en Criotecnología de Alimentos (CIDCA- CCT CONICET). Facultad de Ciencias Exactas, Universidad Nacional de La Plata, 47 y 116 (1900), La Plata, Argentina
Analía G. Abraham
Affiliation:
Centro de Investigación y Desarrollo en Criotecnología de Alimentos (CIDCA- CCT CONICET). Facultad de Ciencias Exactas, Universidad Nacional de La Plata, 47 y 116 (1900), La Plata, Argentina Area Bioquímica y Control de Alimentos, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, 47 y 115 (1900), La Plata, Argentina
Carla Tironi Farinati
Affiliation:
Laboratorio de Fisiopatogenia, Departamento de Fisiología, Facultad de Medicina, Universidad de Buenos Aires, Buenos Aires, Argentina
Cristina Ibarra
Affiliation:
Laboratorio de Fisiopatogenia, Departamento de Fisiología, Facultad de Medicina, Universidad de Buenos Aires, Buenos Aires, Argentina
Graciela L. De Antoni
Affiliation:
Centro de Investigación y Desarrollo en Criotecnología de Alimentos (CIDCA- CCT CONICET). Facultad de Ciencias Exactas, Universidad Nacional de La Plata, 47 y 116 (1900), La Plata, Argentina Cátedra de Microbiología, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, 47 y 115 (1900), La Plata, Argentina Comisión de Investigaciones Científicas de la Provincia de Buenos Aires (CIC-PBA), Buenos Aires, Argentina
*
*For correspondence; e-mail: emilianok@cidca.org.ar
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Abstract

Kefir is a fermented-milk beverage originating and widely consumed in the Caucasus as well as in Eastern Europe and is a source of bacteria with potential probiotic properties. Enterohaemorrhagic Escherichia coli producing Shiga toxin is commonly associated with food-transmitted diseases; the most prevalent serotype causing epidemics is Esch. coli O157:H7. The aim of this study was to evaluate the antagonism of Lactobacillus plantarum isolated from kefir against the action on Vero cells of supernatants of the Esch. coli O157:H7 strain 69160 expressing the type-II Shiga toxin (Stx2) and to study the role of the Lactobacillus cell wall in that inhibition. Spent culture supernatants of Esch. coli O157:H7 strain 69160 led to cytotoxic effects on cultured eukaryotic cells as evidenced by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium-bromide–cleavage assay or by lactate-dehyrogenase release. Lb. plantarum CIDCA 83114 reduced the cytotoxic activity of Stx present in strain-69160 supernatants, and this protection was markedly higher than those of Lactobacillus kefir CIDCA 83113 and 8348 and Lb. delbrueckii subsp. bulgaricus CIDCA 333. This antagonism of cytotoxicity was mimicked by Lb. plantarum cell walls but was reduced after heating or protease treatments, thus indicating a protein or peptide as being involved in the protection mechanism. The cell surface of the lactobacilli bound the subunit B of Stx thereby decreasing the cytotoxicity. These interactions could constitute the first step in preventing the damage induced by Esch. coli O157:H7 supernatants, thus representing a valuable means of potentially mitigating the noxious effects of this food pathogen.

Keywords

Lactobacillus plantarumkefircell wallEscherichia coli O157:H7Shiga toxin
Type
Research Article
Information
Journal of Dairy Research , Volume 80 , Issue 1 , February 2013 , pp. 64 - 71
Copyright
Copyright © Proprietors of Journal of Dairy Research 2012

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