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  • New insights into mycotoxin mixtures: the toxicity of low doses of Type B trichothecenes on intestinal epithelial cells is synergistic.

New insights into mycotoxin mixtures: the toxicity of low doses of Type B trichothecenes on intestinal epithelial cells is synergistic.

Toxicology and applied pharmacology (2013-06-06)
Imourana Alassane-Kpembi, Martine Kolf-Clauw, Thierry Gauthier, Roberta Abrami, François A Abiola, Isabelle P Oswald, Olivier Puel
ABSTRACT

Deoxynivalenol (DON) is the most prevalent trichothecene mycotoxin in crops in Europe and North America. DON is often present with other type B trichothecenes such as 3-acetyldeoxynivalenol (3-ADON), 15-acetyldeoxynivalenol (15-ADON), nivalenol (NIV) and fusarenon-X (FX). Although the cytotoxicity of individual mycotoxins has been widely studied, data on the toxicity of mycotoxin mixtures are limited. The aim of this study was to assess interactions caused by co-exposure to Type B trichothecenes on intestinal epithelial cells. Proliferating Caco-2 cells were exposed to increasing doses of Type B trichothecenes, alone or in binary or ternary mixtures. The MTT test and neutral red uptake, respectively linked to mitochondrial and lysosomal functions, were used to measure intestinal epithelial cytotoxicity. The five tested mycotoxins had a dose-dependent effect on proliferating enterocytes and could be classified in increasing order of toxicity: 3-ADON<15-ADON≈DON<NIV≪FX. Binary or ternary mixtures also showed a dose-dependent effect. At low concentrations (cytotoxic effect between 10 and 30-40%), mycotoxin combinations were synergistic; however DON-NIV-FX mixture showed antagonism. At higher concentrations (cytotoxic effect around 50%), the combinations had an additive or nearly additive effect. These results indicate that the simultaneous presence of low doses of mycotoxins in food commodities and diet may be more toxic than predicted from the mycotoxins alone. Considering the frequent co-occurrence of trichothecenes in the diet and the concentrations of toxins to which consumers are exposed, this synergy should be taken into account.

MATERIALS
Product Number
Brand
Product Description

Supelco
Fusarenon X solution, 100 μg/mL in acetonitrile, analytical standard
Sigma-Aldrich
3-Acetyldeoxynivalenol, from Fusarium roseum
Supelco
15-Acetyldeoxynivalenol solution, ~100 μg/mL in acetonitrile, analytical standard
Supelco
3-Acetyldeoxynivalenol
Supelco
3-Acetyldeoxynivalenol solution, ~100 μg/mL in acetonitrile, analytical standard
Supelco
Nivalenol solution, 100 μg/mL in acetonitrile, analytical standard
Sigma-Aldrich
15-O-Acetyl-4-deoxynivalenol from Fusarium graminearum
Sigma-Aldrich
Thiazolyl Blue Tetrazolium Bromide, powder, BioReagent, suitable for cell culture, suitable for insect cell culture, ≥97.5% (HPLC)
Nivalenol in acetonitrile, IRMM®, certified reference material
4-Deoxynivalenol in acetonitrile, IRMM®, certified reference material
Sigma-Aldrich
Thiazolyl Blue Tetrazolium Bromide, 98%
Sigma-Aldrich
Deoxynivalenol
Supelco
Fusarenon X, analytical standard
Supelco
Deoxynivalenol solution, 100 μg/mL in acetonitrile, analytical standard