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2. Consumers, Health, Agriculture and Food Executive Agency THE ROLE OF PROCESSING IN MYCOTOXIN LEVELS IN THE AGRI - FOOD CHAIN - Role of Industrial Processing types (milling, blanching, roasting, etc.) on the fate of mycotoxins in food and feed products 2

3. Consumers, Health, Agriculture and Food Executive Agency Food Processing - Definition 3 Food processing is the transformation of raw ingredients into food or of food into other forms. Food processing dates back to the prehistoric ages when processing implied slaughtering, fermenting, sun drying, preserving with salt and various types of cooking (roasting, smoking, steaming and oven baking). Benefits of food processing include toxin removal, preservation, easing marketing and distribution tasks, increasing food consistency.

4. Consumers, Health, Agriculture and Food Executive Agency Why processing is important? 4 Data produced to ascertain the fate of mycotoxins through the manufacturing chain are necessary:  To interpret the legislation in the correct way and make revisions too  To contribute in setting balanced regulation between raw material and intermediate and finished products  To make aware the stakeholders on the possibility that accepting consignments meeting legislative requirement will be challenged later in the food chain  To perform more accurate dietary intake estimations  To establish the actual risk for consumer and animals in a more comprehensive way

5. Consumers, Health, Agriculture and Food Executive Agency Legislation (Reg. 1881/06) 5 Article 2 - Dried, diluted, processed and compound foodstuffs When applying the maximum levels, the following shall be taken into account:  Changes of the concentration of the contaminant caused by DRYING or DILUTION processes  Changes of the concentration of the contaminant caused by PROCESSING  The relative proportions of the ingredients  The analytical limit of quantification

6. Consumers, Health, Agriculture and Food Executive Agency How to implement article 2? 6

7. Consumers, Health, Agriculture and Food Executive Agency Reg. 1881/2006 – Whereas 34 and 35 7 (34) Maximum levels of Fusarium toxins should be set for unprocessed cereals placed on the market for first-stage processing……….. (35) Since the degree to which Fusarium toxins in unprocessed cereals are removed by cleaning and processing may vary, it is appropriate to set maximum levels for final consumer cereal products as well as for major food ingredients derived from cereals to have enforceable legislation in the interest of ensuring public health protection

8. Consumers, Health, Agriculture and Food Executive Agency Reg. 1881/2006 – Footnote 18 8 The maximum level applies to unprocessed cereals placed on the market for first-stage processing. ‘First- stage processing’ shall mean any physical or thermal treatment, other than drying, of or on the grain. Cleaning, sorting and drying procedures are not considered to be ‘first-stage processing’ insofar no physical action is exerted on the grain kernel itself and the whole grain remains intact after cleaning and sorting. In integrated production and processing systems, the maximum level applies to the unprocessed cereals in case they are intended for first-stage processing. Fusarium toxins

9. Consumers, Health, Agriculture and Food Executive Agency Typical industrial processing 9 o Cleaning o Optical sorting o Milling o Roasting and decaffeination (coffee, dried fruits) o Brewing o Milk – Cheesemaking o Domestic cooking (not industrial) Cereals

10. Consumers, Health, Agriculture and Food Executive Agency Cleaning - I 10 Cereals normally are cleaned before milling. Cleaning is a process intended to remove extraneous material (metal), dust, husks, broken grains, etc. The cleaning process often consists of scouring, aspiration, sieving, and specific gravity separation. Dust, husks, hair and loose superficial particles are blown away by scouring and aspiration. Extensive aspiration can considerably reduce the quantity of mould spores and mycelia on the kernel surface. Sieving and specific gravity separations remove small kernels, kernel debris, weed seeds and other impurities which differ in size and gravity in comparison to normal kernels. Breakdown values are extremely variable as available from the literature since they depend on the physical nature of grains and from the procedures carried out before milling.

11. Consumers, Health, Agriculture and Food Executive Agency Cleaning – II 11 Very effective in reducing mycotoxin contamination. o The efficacy partly depends on the degree of fungal growth within the kernels. The highest fungal infestation and mycotoxin concentrations of cereals are found in the husks, debris, hull and the outer layer of the kernel. Fusarium-damaged kernels are often also of smaller size than sound kernels. Grain screenings (broken pieces of grain, small grains and the dust from the outer layers of the grain) are the residues from the cleaning of cereals. Mycotoxin concentrations are often found to be much higher in the small kernels and broken grain and in the outer layers of the grain. o The screenings are often diluted with more sound grain, but must be considered as a feedstuff at high risk for mycotoxin contamination. They are cheap and mostly used for ruminants, but their use as a feedstuff should be avoided.

12. Consumers, Health, Agriculture and Food Executive Agency Cleaning effects 12 Type of cleaning MycotoxinCommodityReduction (%) Normal cleaning, Brushing deoxynivalenolwheat50-86 Fractioning by gravity separator deoxynivalenolwheatConcentration in the least dense fractions Dehullingdeoxynivalenol, zearalenone and T-2 toxin wheat, barley, rye and oats 40-100 Polishing and pearling deoxynivalenol, zearalenone barley56 Normal cleaning ochratoxin AWheat and barley No reduction

13. Consumers, Health, Agriculture and Food Executive Agency Optical sorting 13 Very effective in removing Fusarium-contaminated grain kernels As far as aflatoxins, effective up to a contamination level of 40 ug/kg

14. Consumers, Health, Agriculture and Food Executive Agency Wheat milling process 14 Wheat grain Bran shorts Flour middlingsSemolina Foilage waste Peeled wheat Pasta Clean wheat Bran 1 Bran 2 Bran 3

15. Consumers, Health, Agriculture and Food Executive Agency Reduction of DON (%) - wheat grain 15

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17. Consumers, Health, Agriculture and Food Executive Agency Overall reduction from wheat grain to pasta 17 80 %

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19. Consumers, Health, Agriculture and Food Executive Agency 19 Corn milling process - Fumonisins

20. Consumers, Health, Agriculture and Food Executive Agency Coffee roasting – Ochratoxin A 20 1 st stage- Drying Endothermic process. T=205°C 2 nd stage. First crack, T=220°C 3 rd stage. Pyrolisis, T=220°C 4 th stage. Second pyrolisis, T=230°C

21. Consumers, Health, Agriculture and Food Executive Agency 21 Effect of coffee roasting on the fate of OTA

22. Consumers, Health, Agriculture and Food Executive Agency OTA in wine 22 The beginning of the fungal infection occurs in the vineyard on the outer parts of the grape. The OTA present in grapes to a certain degree is released to the juice during crushing. Higher levels of OTA are found in red wines as a result of the longer contact of the must with the skins of the grapes Alcoholic and malo-lactic fermentation cause a reduction in OTA in the wine. Red >> white wines

23. Consumers, Health, Agriculture and Food Executive Agency BREWING 23 Aflatoxins and Ochratoxin A are relatively stable at boiling temperatures of the mash cooking step, but are more sensitive to mash malting (protein hydrolysis), wort boiling, and final fermentation, with removal of 12 to 27%, 20 to 30%, and 20 to 30%, respectively in these steps. Chu et al. (1975) Another study revealed that about a 2–13% reduction of ochratoxin A, 3–28% of FB 1 and 9–17% of FB 2 occurred during the fermentation of wort to which ochratoxin A and fumonisins B 1 and B 2 had been added. Scott et al. (1995)

24. Consumers, Health, Agriculture and Food Executive Agency Cheesemaking – Aflatoxin M1 24 AFM1 is predominantly associated with casein, so that cheese curd contains a higher concentration than whey. For aflatoxin M1, the IARC Committee concluded that there is sufficient evidence that aflatoxin M1 is a genotoxic carcinogen; its carcinogenic potency is estimated to be approximately 10 times lower than aflatoxin B1. AFB1: (Group 1); AFM1: (Group 2B) Studies showed that the concentration of AFM1 is about 3 fold higher in many soft cheeses and about 5 fold higher in hard cheeses than in milk. Some studies demonstrated that cheese ripening and proteolysis of casein increases the recovery of AFM1 from naturally- contaminated milk; proteolysis may affect hydrophobic regions on casein associated molecules releasing AFM1. Prandini, 2002

25. Consumers, Health, Agriculture and Food Executive Agency Conversion AFB1 – AFM1 25 Ingesting AFB1, mammals excrete an aliquot as AFM1 in milk (1-3% of the ingested toxin) The amount of excreted AFM1 can differ depending from individual factors and lactation phases Carry-over is 3,3-3,5 higher at the beginning of lactation and is proportional to the productive level For a milk production of 20 and 46 kg/die the excretion of AFM1 in milk is around 2 and 5,7%, respectively (high individual variability)

26. Consumers, Health, Agriculture and Food Executive Agency AFMI in dairy products 26

27. Consumers, Health, Agriculture and Food Executive Agency 27 No statistical significative variation along the ripening period. The trend was alternate throughout 12 products. Authors conclude that on average AFM1 level remains constant.

28. Consumers, Health, Agriculture and Food Executive Agency AFM1 Conversion factors from milk to dairy products 28 ProductConversion factor Raw Milk1 Yogurth1 Cheese medium ripening time4,5 Grana-like5,5 Mozzarella3,3 Goat cheese2,4 Ricotta from goat whey3 Ricotta from mozzarella whey1,2

29. Consumers, Health, Agriculture and Food Executive Agency Patulin in apple derived products 29 Sorting out of damaged fruits Minimisation of the risk caused by physical damage Pressed for juicing as soon as possible after picking the raw fruits. Mould growth normally occurs in a warm environment. Rapid cooling and maintenance of store atmosphere conditions will improve fruit condition. Stored apples should be examined regularly, at least once a month, for rot levels Ideally fruit should be loaded and cooled to less than 5 °C in three to four days and to optimum temperatures within a further two days. The juice should preferably be chilled to < 5 °C and maintained chilled as well kept under ultra-low oxygen conditions until it is concentrated, packaged or pasteurized. Most juice will be heat processed to ensure destruction of enzymes and spoilage organisms. It must be recognized that whilst such processes will generally destroy fungal spores and vegetative mycelium the process conditions will not destroy any patulin which is already present.

30. Consumers, Health, Agriculture and Food Executive Agency How does consumer have to behave? 30 Being mycotoxins strongly heat stable compounds, cooking temperatures are UNABLE to destroy these hazards Mouldy foods should not be consumed (mouldy spots in apples, cheeses, bread, jam should be removed before the consumption but it should be better to avoid their consumption) The use of microwave oven in domestic food preparation seems to be quite effective in reducing in large extent mycotoxin concentration in the commodity

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