EFS 2013 [Compatibility Mode]

The type of interaction between type B trichothecenes on the intestine varies with the dose I. Alassane-Kpembi, T. Gauthier, F. Lyazrhi, R. Abrami, F. A. Abiola, M. Kolf-Clauw, O. Puel and I.P. Oswald

INRA - Toulouse, FRANCE

12th European19 Fusarium Seminar Bordeaux, janvier 2012 12-16 may 2013 Bordeaux, France

AL I M E N TAT I O N AGRICULTURE ENVIRONNEMENT

Mycotoxin co-contamination: A food/feed safety issue 48% 19% 33%

Fungi produce several mycotoxins simultaneously. Feed and food may be contaminated by several fungi. inf. LOD

1 mycotoxin

Human and animal meals are made up of multiple raw materials.

Co-contamination by several mycotoxins is the RULE, not the exception.

>1 mycotoxin

Toxins 2012, 4(9), 663-675

A Three-Year Survey on the Worldwide Occurrence of Mycotoxins in Feedstuffs and Feed

Inês Rodrigues and Karin Naehrer Number of samples: 7049

Most toxicity studies addressed the effect of mycotoxins when present separately. Combined effects data are needed for an update of safety regulations. 12th European Seminar Bordeaux,Fusarium 19 janvier 2012 12-16 may 2013 Bordeaux, France

AL I M E N TAT I O N AGRICULTURE ENVIRONNEMENT

.

Ocurrence of mycotoxins from Fusarium spp in European cereals Fusarium mycotoxins are frequent contaminants of European cereals. Type B trichothecenes take an important place among these contaminants. Mycotoxins

Main food contaminated (% of positive samples)

Type B trichothecenes - Deoxynivalenol - Nivalenol - 3-acetyldeoxynivalenol

Maize (89%), wheat (61%) Maize (35%), oats (21%), wheat (14%) Maize (27%), wheat (8%)

Type A trichothecenes - T-2 toxin - HT-2 toxin

Maize (28%), wheat (21%), oats (21%) Oats (41%), corn (24%), rye (17%)

Zearalenone

Maize (79%), wheat (30%)

Fumonisins - Fumonisin B1 - Fumonisin B2

Maize (66%) Maize (51%)

12th European Seminar Bordeaux,Fusarium 19 janvier 2012 12-16 may 2013 Bordeaux, France

AL I M E N TAT I O N AGRICULTURE ENVIRONNEMENT

(SCOOP, 2003)

Slide 3 LG1

tu devrais mettre en évidence les mycotoxines que tu as étudié dans ton étude dans le tableau...plus évident pour les personnes qui t'écoutent... lguzylack; 18/01/2012

Intestinal mucosa as a mycotoxin target Intestinal mucosa is the first, but also the main barrier against dietary exposure to contaminants.

Following ingestion of contaminated food, intestinal epithelial cells may be exposed to high concentration of mycotoxins, potentially affecting intestinal functions. 12th European Seminar Bordeaux,Fusarium 19 janvier 2012 12-16 may 2013 Bordeaux, France

AL I M E N TAT I O N AGRICULTURE ENVIRONNEMENT

Mycotoxins affect intestinal health A number of intestinal functions are well-known target for mycotoxin effects Deoxynivalenol Fumonisine B1 Patulin Ochratoxin

Nutrients Absorption

Deoxynivalenol Ochratoxin Fumonisin B1 Aflatoxin B1 T-2 toxin

Cell Proliferation

(Bouhet and Oswald, 2005)

12th European Seminar Bordeaux,Fusarium 19 janvier 2012 12-16 may 2013 Bordeaux, France

Ochratoxin Fumonisin B1 Patulin Deoxynivalenol

Barrier Function

Deoxynivalenol Fumonisin B1 Gliotoxin Patulin

Cytokines

Intestinal epithelium AL I M E N TAT I O N AGRICULTURE ENVIRONNEMENT

Deoxynivalenol Nivalenol T2-toxin

Immuno globulin A

Aim of the study Assess the combined effects of DON, NIV and their acetyl derivatives (3–ADON, 15–ADON and FX) in terms of additive, antagonistic or synergistic toxicity against proliferative human intestinal cells.

Trichothecenes

R1

R2

R3

R4

Deoxynivalenol (DON)

OH

H

OH

OH

3-acetyldeoxynivalenol (3-ADON) O-CO-CH3 H

OH

OH

15-acetyldeoxynivalenol (15ADON)

OH

H

O-CO-CH3 OH

Nivalenol (NIV)

OH

OH

OH

OH

Fusarenon-X (FX)

OH

O-CO-CH3 OH

OH

12th European Seminar Bordeaux,Fusarium 19 janvier 2012 12-16 may 2013 Bordeaux, France

AL I M E N TAT I O N AGRICULTURE ENVIRONNEMENT

(Radtke and Clevers, 2005)

Comparative cytotoxicity of Type B trichothecenes Experimental design Proliferating human intestinal Caco-2 cells seeded in 96well plates and incubated for 24 hours

48 hour- exposure to graded levels of mycotoxin - DON or 3-ADON or 15-ADON or NIV : 0 – 7µM

Neutral Red Lysosomial activity

- FX 0 – 0.12µM

MTT Mitochondrial activity

Two cytotoxicity assays performed - The MTT test -Bordeaux, The Neutral Red uptake 19 janvier 2012 test

AL I M E N TAT I O N AGRICULTURE ENVIRONNEMENT

Comparative cytotoxicity of Type B trichothecenes Results NeutralNeutral red uptake Caco-2 48h redtest uptake test

MTT test Caco-2 48h MTT test DON

100

15-ADON NIV FX 3-ADON NIV

15-ADON DON 3-ADON

NIV 15-ADON

80

Cell viability (%)

80

Cell viability (%)

DON

100

15-ADON DON 3-ADON

FX 60

40

20

FX

3-ADON NIV

FX 60

40

20

0

0 0

2

4

6

8

0

Mycotoxin concentration (µM)

MTT

4 Estimation de l’6 IC50

8

Mycotoxin concentration (µM)

IC50(µM) Mycotoxin

2

Neutral red

The two assays led to similar cytotoxic pattern with close IC50 values. The mean IC50 values ranged from 20 nM to 3µM.

DON

1.39 ± 0.07

a

1.19 ± 0.29

a

3-ADON

2.94 ± 0.45

ab

1.99 ± 0.44

b

15-ADON

1.47 ± 0.28

abc

For the overall ranking, mycotoxins A L I Mcytotoxicity E N T A T I O Nfor proliferating enterocytes 0.69 ± 0.16 cd A G R I C U L T U was R E in the increasing order 3-ADON