MYCOTOXINS AND CANCER
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1 International Conference MYCOTOXINS AND CANCER March 24-25, 2016 Marrakesh
2 Mycotoxins in Moroccan Foods and Feeds State of the Art and Challenges A. TANTAOUI ELARAKI & A. ZINEDINE
3 Mycotoxin? 3
4 Substances produced by fungi (moulds) in foods and feeds and presenting specific toxicity to man and animals 4
5 Historical background 5
6 6
7 Ergotisme: feu de la Saint Antoine ou Mal des Ardents 7
8 Agent causal - XVIIth century : bread made with rye flour - This rye was moulded with Claviceps purpurea 8
9 Claviceps purpurea on wheat Claviceps purpurea on rye
10 ATA (Alimentary Toxic Aleukia) - Graet number of humans killed in East Europe (former USSR, 1930 & 1940), Central Europe, United States, Finland & China - Mycotoxins involved : Trichothecenes, produced by Fusarium fungi 10
11 Discovery of aflatoxins - UK (1960): turkeys et duvcks (Christmas); - «Turkey-X disease»; - Peanut flour (Brazil):. Aspergillus flavus spores. Toxic substance involved Aflatoxine 11
12 Since 1960, fast development - Great number of toxigenic fungal species recognized - more than 400 mycotoxins identified - Few are really worrying 12
13 Some of the most worrying mycotoxins Mycotoxin group Mycotoxins Conditions Aflatoxines (AF) AFB1, AFB2, AFG1, AFG2 Tropical & Subtropical Climates Ochratoxins (OT) OTA, OTB, OTC, etc. Mild Climates storage Zearalenone Zearalenone Moulds all over the World Trichothecenes Vomitoxin,Nivalenol Deoxynivalenol Fusarenone X T2 & HT2 toxines, Diacetoxyscirpenol Fumonisins Fumonisins Moulds all over the World Mild & Hot Climates 13
14 Toxinogenic Fungi 14
15 Fungal genera Very important important Aspergillus Penicillium Fusarium Stachybotrys Alternaria Trichothecium To remember Claviceps 15
16 Penicillium Stipe = conidiophore Penicillium citrinum Penicillium verrucosum
17 Aspergillus Aspergillus ochraceus Têtes conidiennes d Aspergillus a: monsériée b: bisériée Aspergillus sp. Aspergillus flavus
18 Fusarium Macroconidies et microconidies de Fusarium Fusarium verticilloides Fusarium moniliforme
19 Chemical Nature of Mycotoxins 19
20 Aflatoxins 20
21 Ochratoxins 21
22 Citrinin Patulin
23 Zearalenone
24 Trichothecenes 24
25 Fumonisins
26 Toxicity 26
27 Man & animals Acute Toxicity High quantities of de toxin ingested within short time Chronical Toxicity Low quantities ingested during long period of time Accumulative effect 27
28 Acute Toxicity rare in Man Before 1960: ergotism, ATA, etc (Kenya): humans killed - Epidemiological survey (CDC, Atlanta, USA): aflatoxins in corn & other grains (1000 ppb > 250 fold tolerable limits)
29 Tremorgenic activity Penitrem Penicillium crustosum Encephalomalacia Fumonisin Fusarium moniliforme Hepatic necrosis Cancer Sterigmatocystin Aflatoxins Aspergillus flavus Aspergillus versicolor Slime syndroma Lung & Brain Mycotoxins Slaframin Hemorrhage Patulin Rhizoctonia leguminicola Plusieurs espèces Liver Hemorrhage Nephrotoxicity Rubratoxin Ochratoxin Penicillium rubrum P. verrucosum; A. ochraceus Œstrogen Zéaralénone F. graminearum Intestinal unrest Austidiol Aspergillus ustus Diversity of Mycotoxicological Syndromas 29
30 Target Organs for Mycotoxins Liver Kidney Nerv. System Aflatoxins * * * Ochratoxins * * * Toxins Trichotecenes * Tremorgenic * Cyclopiazonic acid Zearalenone * * * Endoc. glands * Skin Immune System * * * * * 30
31 Conditions de toxinogenèse 31
32 Milieu (substrat); Paramètres de l environnement (température, stress hydrique, HR de l air); Autres paramètres: - Lésions mécaniques (récolte); - Attaque par insectes; - Présence d autres moisissures, 32
33 Aspergillus flavus sur maïs 33
34 Mycotoxin Metabolization 34
35 - Transformation (metabolization) - Rejection : faeces, urine, saliva, milk, eggs (metabolized or not) - Accumulation in organs: liver, kidneys, etc. (metabolized or not) 35
36 Distribution of a mycotoxin & its metabolites in the organism Digestive tract T (destruction?) Food T Milk T+M Faeces T Muscle T+M Blood T+M Eggs T+M T: mycotoxin M: métabolite Kidney T+M Other Viscera T+M Liver T M Urine T+M
37 Animal Food Contamination with Mycotoxins - Milk - Eggs - Meat - Giblets
38 Aflatoxin B1 Metabolized in Aflatoxin M1 (rejected in milk) 38
39 Fate of Mycotoxins throw technological treatments 39
40 - Traitements thermiques (chaleur) Froid (réfrigération et congélation) Extraction Fermentation Raffinage des huiles brutes Baking 40
41 Heat Stability of aflatoxin M1 Pasteurization conditions Destruction rate (%) 62 C-30 min 70 C-30 min 71 C-40 sec 72 C-45 sec 74 C-30 min 75 C-40 sec 77 C-16 sec 80 C-45 sec ,9+11a; 6 à 13b Total 0-64 a: artificially contaminated milk; b: naturally contaminated milk. 41
42 Vegetable Oil refining - Soda Neutralization à la soude: aflatoxin destruction - Virgin Olive oil (not refined): danger if mould attack on olives 42
43 Moyens de lutte 43
44 - Détoxification - Adsorption - Prévention: au champ, stockage, transport, etc. 44
45 Conclusion 45
46 Organic Food Myth Fresh food Myth destroyed A mouldy food can be non toxic A non mouldy food can be toxic Remain watchful 46
47 Your aflatoxin, would you like it with rice or with pasta?
48 What about Morocco? 48
49 Data on food and feed contamination: sparse and insufficient Lack of epidemiological studies 49
50 Animal Feeds 50
51 1945: Pig poisoning with mouldy feeds (Ninard & Hinterman) 1979: 640 dairy cows vaches killed (Province of Nador): mouldy wheat bran 1991: 216 equines killed (Province of Gharb): mouldy straw
52 Stachybotryotoxicosis on Equines, Gharb (1991) Stachybotris atra 18
53 Animal Feeds Products Analysed AFB1 OTA Published in Soja cake Sunflower cake Compound Feed Poultry Feed Poultry Feed % (20 ppb) Poultry Feed 70-30% (30 ppb) Non researched
54 Human Food
55 Aflatoxin M1 in Pasteurized Milk 7 % over European LMR 55
56 Aflatoxin M1 in Milk Argentina Brazil Corea Spain India Iran Italy Japan Koweit Libiya Portugal Turkey contamination rate % 56
57 Aflatoxins in dry fruits Products Analysed Contaminated Limit Values (ppb) Référ. Peanuts Peanuts Almonds Nuts Pistachio Raisins Raisins Dried Figs Prunes Dates * 820** 18-30* 20* 20* Total Tantaoui Elaraki et al., Actes Inst. Agron. Vét., 1994, 14, Tantaoui Elaraki et al., Actes Inst. Agron. Vét., 2000, 20, * μg/kg Aflatoxin B1 ** μg/kg total aflatoxins B1+B2+G1+G2 57
58 Olives & Olive Oil Aflatoxin B1 Ochratoxin A Products Analysed Positive Limit Values (μg/kg) Positive Limit Values (μg/kg) Greek Style Black Olives Virgin Olive Oil Tantaoui Elaraki & Le Tutour, Oléagineux, 1985, 40,
59 Aflatoxinsin spices 59
60 Grains and derivatives
61 Aflatoxin B1, Ochratoxin A & Zearalenone Aflatoxin B1 Products analysed Positive Level Wheat Barley Corn Wheat bran * - Total 286 * μg/kg 1 18* Ochratoxin A & Zearalenone Positive Products analysed OTA ZEA Wheat Barley Corn Wheat bran * Total * 1,13 2,83 μg/kg 61 Tantaoui Elaraki et al., Actes Inst. Agron. Vét., 1994, 14, 11-6
62 Fumonisin B1, Ochratoxin A & Zearalenone Grains 62
63 Deoxynivalenol (vomitoxin) Ennouari et al. Food Control (2012) 63
64 Ochratoxin A in Bread 64
65 - % of contamination: 48% ( ng/g) > - 26 % samples over European LMR (3ng/g) - OTA in bread is due to flour contamination 65
66 - Average contamination : 13 ng/g - Average Daily Bread Consumption : 577 g - For an adult (60 kg): DI = 126 ng/kg pc > - 9 fold > TDI (14 ng/kg bw) : FAO/WHO Committee (2001) - 7 fold TDI (17.1 ng/kg bw) : EFSA (2006) 66
67 - Potential Exposure of Moroccans to OTA through bread consumption - Exposure probably responsible for chronical renal failure (4000 new cases/year) - Hypothesis to confirm: epidemiological studies
68 Emergent Mycotoxins 68
69 Enniatins Number of samples Positive to Total Enniatins ENA (mg/kg) ENA1 (mg/kg) Meanb Maxc Meanb Maxc a Wheat Corn nd nd Barley 8 7 nd nd a: Enniatins A, A1, B & B1; b: mean value in positive samples; c: maximum value Zinedine A. et al., Food Control, 22 (2011), 1-5
70 Beauvericin & Fusaroprolferin Number of samples Beauvericin (mg/kg) Fusaroproliferin (mg/kg) Positive Meana Maxb Positive Meana Maxb Wheat Corn Barley ND ND ND a: mean value in positive samples; b: maximum value Zinedine A. et al., Food Control, 22 (2011), 1-5
71 Breakfast & Infant Cereals Mahnine N. et al. Food Chemistry, 124 (2011),
72 72
73
74 Emerging Fusarium mycotoxins in Rice Number of samples Positive to Total Enniatinsa ENA (mg/kg) Meanb Maxc ENA1 (mg/kg) Meanb Maxc Rabat nd Casablanca nd Kénitra Errachidia nd Tangiers Mohammedia a: Enniatins A, A1, B & B1; b: mean value in positive samples; c: maximum value nd nd nd Sifou A. et al., Food Control, 22 (2011),
75 Norvège, Finlande: ENB > ENB1 > ENA1 > ENA Italie, USA: FUS & BEA > ENs Maroc, Espagne & Tunisie: ENA1 > ENB1 > ENB > ENA Influence des conditions climatiques?
76 Législation marocaine?
77
78 Quelques spécificités du Maroc 78
79 Trituration d olives fortement moisies dans les mâasras traditionnelles
80
81 Distribution de pain moisi aux vaches laitières Photo M. Majdi 81
82 Réglementation SSA Mycotoxines 82
83 83
A. Zinedine1 & A. Tantaoui-Elaraki2
12-14th October 2011 Istanbul, Turkey Mycotoxins in Moroccan Foods: Current Situation and Main Perspectives A. Zinedine1 & A. Tantaoui-Elaraki2 1: National Hygiene Institute, Rabat 2: SUP AGRO, Casablanca
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