Deoxynivalenol and its metabolite deepoxy-deoxynivalenol: multi-parameter analysis for the evaluation of cytotoxicity and cellular effects

Alexandra Springler,Sabine Hessenberger,Veronika Nagl,Nicole Reisinger,Gerd Schatzmayr,Elisabeth Mayer,Corinna Kern

doi:10.1007/s12550-016-0260-z

Alexandra Springler, Sabine Hessenberger + Show 5 more

Open Access

https://doi.org/10.1007/s12550-016-0260-z

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Abstract

The mycotoxin deoxynivalenol (DON) contaminates agricultural commodities worldwide, posing health threats to humans and animals. Associated with DON are derivatives, such as deepoxy-deoxynivalenol (DOM-1), produced by enzymatic transformation of certain intestinal bacteria, which are naturally occurring or applied as feed additives. Using differentiated porcine intestinal epithelial cells (IPEC-J2), we provide the first multi-parameter comparative cytotoxicity analysis of DON and DOM-1, based on the parallel evaluation of lysosomal activity, total protein content, membrane integrity, mitochondrial metabolism and ATP synthesis. The study investigated the ability of DON and—for the first time of its metabolite DOM-1—to induce apoptosis, mitogen-activated protein kinase (MAPK) signalling, oxidative events and alterations of mitochondrial structure in porcine intestinal epithelial cells (IECs). The degree of DON toxicity strongly varied, depending on the cytotoxicity parameter evaluated. DON compromised viability according to the parameters of lysosomal activity, total protein content and membrane integrity, but increased viability according to assays based on mitochondrial metabolism and ATP synthesis. DON induced expression of cleaved caspase-3 (maximum induction 3.9-fold) and MAPK p38 and p42/p44 (maximum induction 2.51- and 2.30-fold, respectively). DON altered mitochondrial morphology, but did not increase intracellular ROS. DOM-1-treated IPEC-J2 remained unaffected at equimolar concentrations in all assays, thereby confirming the safety of feed additives using DON- to DOM-1-transforming bacteria. The study additionally highlights that an extensive multi-parameter analysis significantly contributes to the quality of in vitro data.

Highlights

Mycotoxins are fungal secondary metabolites, which contaminate up to approximately 70% of all agricultural commodities (Streit et al 2012)
Due to the oral route of DON exposure and the fact that intestinal epithelial cells (IECs) are confronted with high mycotoxin concentrations upon consumption of contaminated food or feed (Bouhet and Oswald 2005; Bracarense et al 2012), the present study examines the effect of DON on viability and function of cells forming this organ system
In vitro cytotoxicity assays facilitate the detection of many mycotoxins, which potentially impair the biochemical activity of animal or human cells

Summary

Introduction

Mycotoxins are fungal secondary metabolites, which contaminate up to approximately 70% of all agricultural commodities (Streit et al 2012). The fusariotoxin deoxynivalenol (DON) is one of the most prevalent of such food- and feed-associated contaminants (EFSA 2013; Rotter et al 1996). Its toxicity depends primarily on species-specific susceptibility, as well as on severity and duration of exposure. Acute DON exposure in pigs can trigger diarrhoea, emesis, leucocytosis, haemorrhage, endotoxemia and, shock-like death. Chronic low-dose exposure may result in growth retardation and immunological impairments (Pestka 2010a, b; Rotter et al 1996). Pigs, whose diet is largely composed of cereal-rich feed, exhibit high DON susceptibility (Prelusky et al 1994)

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