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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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References

Abraham, AG, De Antoni, G & Añon, MC 1993 Proteolytic activity of Lactobacillus bulgaricus grown in milk. Journal of Dairy Science 76 1498–1505Google Scholar
Banatvala, N, Griffin, PM, Greene, KD, Barrett, TJ, Bibb, WF, Green, JH & Wells, JG 2001 The United States national prospective hemolytic uremic syndrome study: microbiologic, serologic, clinical, and epidemiologic findings. Journal of Infectious Diseases 183 1063–1070Google Scholar
Boekhorst, J, Wels, M, Keerebezem, M & Siezen, R 2006 The predicted secretome of Lactobacillus plantarum WCFS1 sheds light on interactions with its environment. Microbiology 152 3175–3183CrossRefGoogle ScholarPubMed
Boerlin, P, Mc Ewen, SA, Boerlin-Petzold, F, Wilson, JB, Johnson, RP & Gyles, CL 1999 Association between virulence factors of shiga toxin-producing Escherichia coli and disease in humans. Journal of Clinical Microbiology 37 497–503Google Scholar
Carey, CM, Kostrzynska, M, Ojha, S & Thompson, S 2008 The effect of probiotics and organic acids on shiga-toxin 2 expression in enterohemorrhagic Escherichia coli O157:H7. Journal of Microbiological Methods 73 125–132CrossRefGoogle ScholarPubMed
Garrote, G, Abraham, AG & De Antoni, G 2000 Inhibitory power of kefir: the role of organics acids. Journal of Food Protection 63 364–369CrossRefGoogle Scholar
Garrote, G, Abraham, A & De Antoni, G 2001 Chemical and microbiological characterization of kefir grains. Journal of Dairy Research 68 639–652CrossRefGoogle ScholarPubMed
Garrote, GL, Delfederico, L, Bibiloni, R, Abraham, AG, Pérez, PF, Semorile, L & De Antoni, GL 2004 Lactobacilli isolated from kefir grains: evidence of the presence of S-layer proteins. Journal of Dairy Research 71 222–230Google Scholar
Golowczyc, MA, Mobili, P, Garrote, GL, Abraham, AG & De Antoni, GL 2007 Protective action of lactobacillus kefir carrying s-layer protein against salmonella enterica serovar enteritidis. International Journal of Food Microbiology 118 264–273Google Scholar
Golowczyc, MA, Gugliada, MJ, Hollmann, A, Delfederico, L, Garrote, GL, Abraham, AG, Semorile, L & De Antoni, GL 2008 Characterization of homofermentative lactobacilli isolated from kefir grains: potential use as probiotic. Journal of Dairy Research 75 211–217Google Scholar
Hertzler, S & Clancy, S 2003 Kefir improves lactose digestion and tolerance in adults with lactose maldigestion. Journal of the American Dietetic Association 103 582–587Google Scholar
Hong, W-S, Chen, H-C, Chen, Y-P & Chen, M-J 2009 Effects of kefir supernatant and lactic acid bacteria isolated from kefir grain on cytokine production by macrophage. International Dairy Journal 19 244–251Google Scholar
Hugo, AA, Kakisu, E, De Antoni, GL & Perez, PF 2008 Lactobacilli antagonize biological effects of enterohaemorrhagic Escherichia coli in vitro. Letters in Applied Microbiology 46 613–619CrossRefGoogle ScholarPubMed
Kakisu, E, Abraham, AG, Pérez, P & De Antoni, G 2007 Inhibition of bacillus cereus by kefir grains fermented milk. Journal of Food Protection 70 2613–2616CrossRefGoogle Scholar
Kakisu, E, Irigoyen, A, Torre, P, De Antoni, GL & Abraham, AG 2011 Physicochemical, microbiological and sensory profiles of fermented milk containing probiotic strains isolated from kefir. Journal of Dairy Research 78 456–463Google Scholar
Kim, Y, Han, K-S, Imm, J-Y, Oh, S, You, S, Park, S & Kim, S-H 2006 Inhibitory effects of lactobacillus acidophilus lysates on the cytotoxic activity of shiga-like toxin 2 produced from Escherichia coli O157:H7. Letters in Applied Microbiology 43 502–507Google Scholar
Kitov, P, Sadowska, J, Mulvey, G, Armstrong, G, Ling, H, Pannu, N, Read, R & Bundle, D 2000 Shiga-like toxins are neutralized by tailored multivalent carbohydrate ligands. Nature 403 669–672CrossRefGoogle ScholarPubMed
Liu, J-R, Chen, M-J & Lin, C-W 2005 Antimutagenic and antioxidant properties of milk – kefir and soymilk- kefir. Journal of Agricultural and Food Chemistry 53 2467–2474CrossRefGoogle ScholarPubMed
Liu, Z, Shen, T, Zhang, P, Ma, Y & Qin, H 2011 Lactobacillus plantarum surface layer adhesive protein protects intestinal epithelial cells against tight junction injury induced by enteropathogenic Escherichia coli. Molecular Biology Reports 38 3471–3480Google Scholar
Mosmann, T 1983 Rapid colorimetric assay for cellular growth and survivals: application to proliferation and cytotoxicity assays. Journal of Immunological Methods 65 55–63Google Scholar
Neri, P, Nagano, SI, Yokoyama, S-I, Dohi, H, Kabayashi, K, Miura, T, Inazu, T, Sugiyama, T, Nishida, Y & Mori, H 2007 Neutralizing activity of polyvalent Gb3, Gb2 and Galacto-Trehalose models against Shiga toxins. Microbiology and Immunology 51 581–592Google Scholar
Oelschlaeger, T 2009 Mechanisms of probiotic actions–A review. International Journal of Medical Microbiology 300 57–62CrossRefGoogle ScholarPubMed
O'Loughlin, EV & Robins-Browne, RM 2001 Effect of Shiga toxin and Shiga-like toxins on eukaryotic cells Microbes and Infection 3 493–507Google Scholar
Paton, AW, Morona, R & Paton, JC 2001a Neutralization of Shiga toxins stx1, stx2c, and stx2e by recombinant bacteria expressing mimics of globotriose and globotetraose Infection and Immunity 69 1967–1970CrossRefGoogle ScholarPubMed
Paton, JC, Rogers, TJ, Morona, R & Paton, AW 2001b Oral administration of formaldehyde-killed recombinant bacteria expressing a mimic of the Shiga toxin receptor protects mice from fatal challenge with Shiga-toxigenic Escherichia coli. Infection and Immunity 69 1389–1393Google Scholar
Paton, JC & Paton, AW 1998 Pathogenesis and diagnosis of Shiga toxin-producing Escherichia coli infections. Clinical Microbiology Reviews 11 450–479Google Scholar
Pinyon, RA, Paton, JC, Paton, AW, Botten, JA & Morona, R 2004 Refinement of a therapeutic Shiga toxin-binding probiotic for human trials. Journal of Infectious Diseases 189 1547–1555Google Scholar
Rivas, M, Miliwebsky, E, Chinen, I, Deza, N & Leotta, G 2006 Epidemiología del síndrome urémico hemolítico en argentina. diagnóstico del agente etiológico, reservorios y vías de transmisión. Medicina 66 27–32Google Scholar
Schägger, H & von Jagow, G 1987 Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa Analytical Biochemistry 166 368–379Google Scholar
Silva, KR, Rodrigues, SA, Filho, LX & Lima, AS 2009 Antimicrobial activity of broth fermented with kefir grains. Applied Biochemistry and Biotechnnology 152 316–325CrossRefGoogle ScholarPubMed
Smith, WE, Kane, AV, Campbell, ST, Acheson, DW, Cochran, BH & Thorpe, CM 2003 Shiga toxin 1 triggers a ribotoxic stress response leading to p38 and JNK activation and induction of apoptosis in intestinal epithelial cells. Infection and Immunity 71 1497–1504CrossRefGoogle ScholarPubMed
Vinderola, G, Perdigón, G, Duarte, J, Thangavel, D, Farnworth, E & Matar, C 2006 Effects of kefir fractions on innate immunity. Inmunobiology 211 149–156Google Scholar