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Kefir-isolated Lactococcus lactis subsp. lactis inhibits the cytotoxic effect of Clostridium difficile in vitro

Published online by Cambridge University Press:  10 December 2012

Patricia Araceli Bolla ,
Paula Carasi ,
María de los Angeles Serradell  and
Graciela Liliana De Antoni
Patricia Araceli Bolla
Affiliation:
Cátedra de Microbiología, Departamento de Ciencias Biológicas, Facultad de Ciencias Exactas, Universidad Nacional de La Plata (UNLP), 47 y 115, La Plata, Argentina Centro de Investigación y Desarrollo en Criotecnología de Alimentos (CIDCA), CCT-CONICET, 47 y 116, La Plata, Argentina
Paula Carasi
Affiliation:
Cátedra de Microbiología, Departamento de Ciencias Biológicas, Facultad de Ciencias Exactas, Universidad Nacional de La Plata (UNLP), 47 y 115, La Plata, Argentina
María de los Angeles Serradell*
Affiliation:
Cátedra de Microbiología, Departamento de Ciencias Biológicas, Facultad de Ciencias Exactas, Universidad Nacional de La Plata (UNLP), 47 y 115, La Plata, Argentina
Graciela Liliana De Antoni
Affiliation:
Cátedra de Microbiología, Departamento de Ciencias Biológicas, Facultad de Ciencias Exactas, Universidad Nacional de La Plata (UNLP), 47 y 115, La Plata, Argentina Centro de Investigación y Desarrollo en Criotecnología de Alimentos (CIDCA), CCT-CONICET, 47 y 116, La Plata, Argentina
*
*For correspondence; e-mail: [email protected]
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Abstract

Kefir is a dairy product obtained by fermentation of milk with a complex microbial population and several health-promoting properties have been attributed to its consumption. In this work, we tested the ability of different kefir-isolated bacterial and yeast strains (Lactobacillus kefir, Lb. plantarum, Lactococcus lactis subps. lactis, Saccharomyces cerevisiae and Kluyveromyces marxianus) or a mixture of them (MM) to antagonise the cytopathic effect of toxins from Clostridium difficile (TcdA and TcdB). Cell detachment assays and F-actin network staining using Vero cell line were performed. Although incubation with microbial cells did not reduce the damage induced by C. difficile spent culture supernatant (SCS), Lc. lactis CIDCA 8221 and MM supernatants were able to inhibit the cytotoxicity of SCS to Vero cells. Fraction of Lc. lactis CIDCA 8221 supernatant containing components higher than 10 kDa were responsible for the inhibitory activity and heating of this fraction for 15 min at 100 °C completely abrogated this ability. By dot-blot assay with anti-TcdA or anti-TcdB antibodies, concentration of both toxins seems to be reduced in SCS treated with Lc. lactis CIDCA 8221 supernatant. However, protective effect was not affected by treatment with proteases or proteases-inhibitors tested. In conclusion, we demonstrated that kefir-isolated Lc. lactis CIDCA 8221 secreted heat-sensitive products able to protect eukaryotic cells from cytopathic effect of C. difficile toxins in vitro. Our findings provide new insights into the probiotic action of microorganisms isolated from kefir against virulence factors from intestinal pathogens.

Keywords

Lactococcus lactis subsplactiskefirClostridium difficiletoxin
Type
Research Article
Information
Journal of Dairy Research , Volume 80 , Issue 1 , February 2013 , pp. 96 - 102
Copyright
Copyright © Proprietors of Journal of Dairy Research 2012

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