Comparative effect of DON and plant or microbial biotransformation of DON on gene expression of piglets jejunal explants
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ABSTRACT: Deoxynivalenol (DON) is a frequent mycotoxin in grains, produced by Fusarium fungi, which demonstre multiple side effects such as modulation of immune responses, reduced feed intake and weight gain or impairment of the intestinal barrier function. Among animal species, pigs are the best model for humans and are very sensitive to DON. In wheat, DON can be conjugated to glucose to form DON-3-β-D-glucoside (D3G). Some bacteria isolated from digestive tracts or soil, are also able to de-epoxydize or epimerize DON to metabolites such as deepoxy-deoxynivalenol (DOM-1) or 3-epi-deoxynivalenol (epi-DON). The toxicity of these DON metabolites is poorly documented. By the way of ingestion, the intestine is the first organ exposed to these molecules and so constitute a relevant model. The aim of this study was to compare the intestinal toxicity of three DON metabolites (D3G, DOM-1 and epi-DON) with the one of DON. Intestinal explants from 6 pigs were treated with 10mM DON, D3G, DOM-1 or epi-DON for 4 hours and transcriptomic analysis was performed using an âAgilent Porcinet 60Kâ. Jejunal explants from 4 piglets aged of 5 weeks were sampled and exposed in vitro to differents molecules (DON, D3G, DOM-1 & 3-epi-DON) at 10µM during 4h. Then RNA was extracted.
Project description:Deoxynivalenol (DON) is a frequent mycotoxin in grains, produced by Fusarium fungi, which demonstre multiple side effects such as modulation of immune responses, reduced feed intake and weight gain or impairment of the intestinal barrier function. Among animal species, pigs are the best model for humans and are very sensitive to DON. In wheat, DON can be conjugated to glucose to form DON-3-β-D-glucoside (D3G). Some bacteria isolated from digestive tracts or soil, are also able to de-epoxydize or epimerize DON to metabolites such as deepoxy-deoxynivalenol (DOM-1) or 3-epi-deoxynivalenol (epi-DON). The toxicity of these DON metabolites is poorly documented. By the way of ingestion, the intestine is the first organ exposed to these molecules and so constitute a relevant model. The aim of this study was to compare the intestinal toxicity of three DON metabolites (D3G, DOM-1 and epi-DON) with the one of DON. Intestinal explants from 6 pigs were treated with 10mM DON, D3G, DOM-1 or epi-DON for 4 hours and transcriptomic analysis was performed using an “Agilent Porcinet 60K”.
Project description:Deoxynivalenol (DON) is a type B trichothecene mycotoxin that is commonly found in cereals and grains worldwide. The presence of this fungal secondary-metabolite raises public-health concerns at both the agriculture and food industry level. The toxicity of DON is mainly characterized by its ability to inhibit ribosomal protein biosynthesis. Recently, we have shown that DON has a negative impact on gut integrity, a feature also noticed for Campylobacter (C.) jejuni. We further demonstrated that DON increased the load of C. jejuni in the gut and inner organs. In contrast, feeding the less toxic DON metabolite deepoxy-deoxynivalenol (DOM-1) to broilers reduced the Campylobacter load in vivo. Consequently, it can be hypothesized that DON and DOM-1 have a direct or indirect effect on the growth profile of C. jejuni. The aim of the present study was to further resolve the nature of this interaction in vitro by co-incubation and RNA-sequencing. The co-incubation of C. jejuni with DON resulted in significantly higher bacterial growth rates from 30 h of incubation onwards. On the contrary, the co-incubation of C. jejuni with DOM-1 reduced the CFU counts, indicating that this DON metabolite might contribute to reduce the burden of C. jejuni in birds, altogether confirming in vivo data. Furthermore, the transcriptomic profile of C. jejuni following incubation with either DON or DOM-1 differed. Co-incubation of C. jejuni with DON significantly increased the expression of multiple genes which are critical for Campylobacter growth, particularly members of the Flagella gene family, frr (ribosome-recycling factor), PBP2 futA-like (Fe3+ periplasmic binding family) and PotA (ATP-binding subunit). These organelles are required for pathogenicity-related phenotypes including motility, biofilm formation, host cell interactions, and host colonization, which may explain the high Campylobacter load in the intestine of DON-fed broiler chickens. On the contrary, DOM-1 downregulated the Flagella gene family and upregulated ribosomal proteins. The results highlight the adaptive mechanisms involved in the transcriptional response of C. jejuni to DON and its metabolite DOM-1, based on the following effects: (a) ribosomal proteins; (b) flagellar proteins; (c) engagement of different metabolic pathways. The results provide insight into the response of an important intestinal microbial pathogen against DON and lead to a better understanding of the luminal or environmental acclimation mechanisms in chickens.
Project description:Transcriptional profiling of log and stationary phase S. Typhimurium, comparing untreated controls with Deoxynivalenol treated samples. Each array used labelled cDNA against a common genomic DNA reference. Triplicate arrays were carried out for each of the 4 conditions: untreated log phase, untreated stationary phase, DON treated log phase and DON treated stationary phase
Project description:Trichothecenes, like deoxynivalenol (DON) are secondary metabolites of Fusarium species, and are major pollutants in cereal food and feed products. They inhibit eukaryotic protein synthesis, however with only partly resolved consequences for the cellular homeostasis. Here we report a systematic functional investigation of the effects of DON on exponential phase growing yeast cells using either short doses of DON or continuos treatment with DON.
Project description:Fusarium head blight (FHB) is a major disease of cereal crops caused by the fungus Fusarium graminearum (Fg). FHB affects the flowering heads (or spikes). This study compare the gene expression profile in wheat spikelets from the very susceptible spring wheat cultivar Roblin inoculated with either water (H2O), a Fg strain (GZ3639) producing the mycotoxin deoxynivalenol (+DON), or a GZ3639-derived Fg strain which has been inactivated at the Tri5 locus (-DON).
Project description:Deoxynivalenol (DON) is an important Fusarium toxin of concern for food safety. The inhalation of powder contaminated with deoxynivalenol is possible and may cause lung toxicity. In this study, we investigated the gene expression profile of A549 cells treated with 0.2 microg/mL deoxynivalenol by microarray analysis. In total, 16 genes and 5 noncoding RNAs were significantly affected by deoxynivalenol treatment. 4 samples; 2 samples from nontreated cells and other 2 samples from 0.2 microg/mL DON-treated cells for 24 hours
Project description:The effect of an artificial infection with Fusarium culmorum and an application of deoxynivalenol (DON) on barley spikes of cultivars Chevron and Pedant during flowering was investigated at grain mid-dough stage 10 days after pathogen inoculation (10 dai). Proteomic analysis using a two-dimensional differential gel electrophoresis (2D-DIGE) technique coupled with LC-MS/MS was used to investigate the quantitative or qualitative differences between the experimental variants.
Project description:affy_brachy_2011_11 - affy_brachy_2011_11 - Fusarium graminearum is the causal agent of Fusarium head blight (FHB) of small-grain cereals, including wheat. Besides direct grain losses, this disease is of major concern because of the production by the pathogen of mycotoxins which are hazardous to animals, thus making the grain unfit for food or feed. Major mycotoxins produced by the fungus are trichothecens, including deoxynivalenol (DON). In our laboratory, we use Brachypodium distachyon as a model plant for cereals because of its amenability (short life cycle, numerous genomic and genetic resources, ...). We have recently shown that F. graminearum does induce head blight symptoms on this species and that DON is produced on infected spikes. We have also evidenced that a F. graminearum strain unable to produce DON exhibits reduced virulence on B. distachyon spikes, as previously shown on wheat. The aim of this project is to analyse and compare the plant response to DON producing and non-producing strains of F. graminearum. This analysis will allow to decipher the mechanisms of detoxification set up by the plant and also more specific responses due to the impact of the mycotoxin on plant metabolism and physiology. -Three conditions on B. distachyon spikes: 1-Mock inoculation (Tween 20 0,01%) 2-Inoculation by a F. graminearum wild-type strain 3-Inoculation by a F. graminearum mutant strain, unable to produce DON Spikes were point inoculated with 3ul of either Tween 20 0.01%, wild-type strain or mutant strain (300 spores) and incubated for 96 hours. Six inoculated spikes were collected and pooled for each condition and biological replicate. Three independent biological replicates were conducted.
Project description:affy_brachy_2011_11 - affy_brachy_2011_11 - Fusarium graminearum is the causal agent of Fusarium head blight (FHB) of small-grain cereals, including wheat. Besides direct grain losses, this disease is of major concern because of the production by the pathogen of mycotoxins which are hazardous to animals, thus making the grain unfit for food or feed. Major mycotoxins produced by the fungus are trichothecens, including deoxynivalenol (DON). In our laboratory, we use Brachypodium distachyon as a model plant for cereals because of its amenability (short life cycle, numerous genomic and genetic resources, ...). We have recently shown that F. graminearum does induce head blight symptoms on this species and that DON is produced on infected spikes. We have also evidenced that a F. graminearum strain unable to produce DON exhibits reduced virulence on B. distachyon spikes, as previously shown on wheat. The aim of this project is to analyse and compare the plant response to DON producing and non-producing strains of F. graminearum. This analysis will allow to decipher the mechanisms of detoxification set up by the plant and also more specific responses due to the impact of the mycotoxin on plant metabolism and physiology. -Three conditions on B. distachyon spikes: 1-Mock inoculation (Tween 20 0,01%) 2-Inoculation by a F. graminearum wild-type strain 3-Inoculation by a F. graminearum mutant strain, unable to produce DON Spikes were point inoculated with 3ul of either Tween 20 0.01%, wild-type strain or mutant strain (300 spores) and incubated for 96 hours. Six inoculated spikes were collected and pooled for each condition and biological replicate. Three independent biological replicates were conducted. 9 arrays - Brachypodium; normal vs disease comparison,time course
Project description:To screen cellular effects on Caco-2 cells by deoxynivalenol (DON) and 15-acetyldeoxynivalenol (15ADON), toxin-treated Caco-2 cells were analyzed by DNA microarray. Exposure to either toxin induced up- or downregulation of genes in Caco-2 cells. Upregulation of 1290 and 3238 genes was observed for the DON- and 15ADON-treated groups, respectively, after a 60-min incubation period. This represented a greater than 1.5-fold change relative to the 0-min exposure (control) group. Five hundred and sixty-one genes showed a 1.5-fold upregulation in the both the DON- and 15ADON-treated groups. Similarly, 13 genes and 105 genes were upregulated with a log2 ratio > 2 (4-fold change) in the DON- and 15ADON-treated groups, respectively. The most remarkable gene upregulation corresponded to the inflammatory chemokine, IL-8, which showed a greater than 4-fold upregulation in response to both treatments. Deoxynivalenol and 15-acetyldeoxynivalenol at concentrations of 1 ug/mL were added separately to the AP chamber, and cells were harvested with 0.5% trypsin-EDTA after incubation for 0, 5, 30, and 60 min. Toxin-treated Caco-2 cells were analyzed by DNA microarray to screen for changes in gene expression in response to deoxynivalenol or 15-acetyldeoxynivalenol.