Selection of escherichia coli heat-labile toxin (LT) inhibitors using both the gm1-elisa and the cAMP Vero cell assay

Escherichia coli protein; Escherichia coli; Vero cell; Abrus; Canavalia; Chlorocebus aethiops; Escherichia coli Infections; Fabaceae; Swine Diseases; Vero cell line; Animalia; Acetylgalactosamine; Animals; Bacterial Toxins; Cell Survival; Cercopithecus aethiops; Diarrhea; Enterotoxigenic Escherichia coli; Enterotoxins; Enzyme-Linked Immunosorbent Assay; Escherichia coli Proteins; Galactose; Humans; Lactose; Plant Extracts; Polyphenols; Protein Binding; Receptors, Cell Surface; Swine; Tea; Time Factors; Vero Cells

Abstract :

[en] Weaned piglets are very susceptible to diarrhea caused by enterotoxigenic Escherichia coli. In the past, various natural components were proposed to have beneficial effects by reducing the effects of diarrheal infectious diseases in humans and animals, and thus may represent an alternative for the use of (prophylactic) antibiotics. Alternatives may inactivate enterotoxigenic Escherichia coli heat-labile toxin (LT) by interfering with toxin binding to the cellular receptor GM1. In this study, various plants and other natural substances were tested for inhibitory properties, in the GM1 binding assay, and in the LT-induced cAMP production in Vero cells. The toxic dose of each compound was determined in a cell viability assay, and the highest nontoxic concentrations were used in the GM1 and cAMP assays. Results demonstrated that only d-(+)-galactose, lactose, N-acetyl-d-galactosamine, and two tea extracts were able to inhibit the binding of LT to its GM1 receptor. In the cAMP assay, only the two tea extracts showed inhibitory activity. This shows that d-(+)-galactose, lactose, and N-acetyl-d-galactosamine can indeed inhibit LT binding to GM1 based on structural homology with GM1 in the absence of living cells. However, in the cAMP assay, d-(+)-galactose, and lactose, N-acetyl-d-galactosamine are apparently metabolized to below their effective inhibitory concentration, likely predicting limited practical applicability in vivo. Both tea extracts maintained their activity in the presence of cells. The active compounds in both are probably polyphenols, which are not easily metabolized, and most likely work by aggregating the toxin. In conclusion, the combination of methods used here is a convenient and fast method for preselecting natural substances containing potentially toxin-binding compounds. Furthermore, if antidiarrhea activity is attributed to compounds found inactive here, their activity is unlikely based on interference with toxin binding. © Copyright 2013, Mary Ann Liebert, Inc. 2013.

Disciplines :

Animal production & animal husbandry

Author, co-author :

Verhelst, R.; Nutrition and Health Unit, Department of Biosystems, KU Leuven, Kasteelpark Arenberg 30 Box 2456, 3001 Heverlee, Belgium

Schroyen, Martine ; Université de Liège > Agronomie, Bio-ingénierie et Chimie (AgroBioChem) > Zootechnie

Buys, N.; Livestock Genetics Laboratory, Department of Biosystems, KU Leuven, Heverlee, Belgium

Niewold, T.; Nutrition and Health Unit, Department of Biosystems, KU Leuven, Kasteelpark Arenberg 30 Box 2456, 3001 Heverlee, Belgium

Title :

Selection of escherichia coli heat-labile toxin (LT) inhibitors using both the gm1-elisa and the cAMP Vero cell assay

Publication date :

2013

Journal title :

Foodborne Pathogens and Disease

ISSN :

1535-3141

eISSN :

1556-7125

Publisher :

Mary Ann Liebert, United States

Volume :

Issue :

Pages :

603-607

Peer reviewed :

Peer Reviewed verified by ORBi

Available on ORBi :

since 24 April 2017

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