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1 1 1 Joint Research Centre (JRC) Occurrence of Mycotoxins in Food and Feed Gıda ve Yemlerde Mikotoksinlerin Oluşumu Joerg Stroka IRMM - Institute for Reference Materials and Measurements Geel - Belgium A long lasting history 2 2 ~ "Wailing and writhing men collapsed in the street: others fell over and foamed in epileptic fits whilst some vomited and showed signs of insanity. Many of them shouted "Fire! I'm burning". It was an invisible fire that separated the flesh from the bones and consumed it. Men, women and children died in unbearable agonising pain."

.."Wailing and writhing men collapsed in the street: others fell over and foamed in epileptic fits whilst some vomited and showed signs of insanity.

2 WHO perspective 3 3 Often cited WHO statement: More than 20% of food worldwide are contaminated with mycotoxins WHO nun Görüşü: Dünyadaki gıdaların %20 sinden fazlası mikotoksinlerle kontamine olmuştur. Four Dimensions Dört Boyut 4 4 Occurrence as function of time Oluşumda zamanın etkisi Occurrence can change over time for many reasons Oluşum pekçok sebepten ötürü zaman içinde değişebilir. Occurrence as function of the matrix Oluşumda matriksin etkisi Fungy have a preference for the substrate they grow on Küf... Occurrence as function of production practices Oluşumda üretim yönteminin etkisi e.g. GMP, food processing, storage Örn. GMP, üretim, depolama... Occurrence as function of the environment Oluşumda çevrenin etkisi pre- or post-harvest conditions, humidity, etc hasat öncesi ya da sonrası şartlar, nem vs. 2

Occurrence as function of the matrix Oluşumda matriksin etkisi Fungy have a preference for the substrate they grow on Küf.

3 Humidity THE key factor Nem Anahtar faktör 5 5 Insufficiently dry prior harvest/storage Hasat/Depolama öncesi yetersiz kurutma Insect damage (pre and post harvest) Böcek yeniği Formation of hot-spots (various reasons, also insect damage) Kritik noktaların oluşması ( böcek ısırığı gibi çeşitli nedenler) Penetration of water (e.g. condensation) Ürüne su girmesi Every fungi has its matrix to grow on Her küfün gelişebileceği uygun matriksi vardır 6 6 Most fungi grow / produce mycotoxins only on a limited number of matrices: Çoğu küf mikotoksinleri sadece sınırlı sayıdaki matrikste üretebilirler Penicillium expansum produces patulin mainly in apple juice Penicillium expansum patulini temelde elma suyunda üretir. Fusarium moniliforme produces fumonisins mainly in maize Fusarium moniliforme fumonisinleri genelde mısırda üretir Some matrices are more susceptible hosts than others: Bazı matrisler diğerlerinden daha hassas taşıyıcılardır. Maize is subject to all kinds of fungal infestations and many mycotoxins can be found on maize. Mısır bütün küf kaynaklı hastalıklara konu olur ve pek çok mikotoksin mısırda bulunabilir. Fungal growth on oats is more limited (mainly T-2 and HT-2 toxins) Yulafta küf gelişimi daha sınırlıdır (genelde T-2 ve HT-2 toksinleri) 3

) Kritik noktaların oluşması ( böcek ısırığı gi

4 Legislation as result for occurrence data Oluşma verilerinin sonucu olarak mevzuat 7 7 Most mycotoxins that are considered as a health risk for humans and animals will be regulated over time in the products of concern: İnsan ve hayvanlar için sağlık riski taşıyan birçok mikotoksin için zaman içerisinde ilgili ürünlere göre mevzuatlar düzenlenmktedir. Regulated in the EU: Aflatoxin (B1, B2, G1, G2 and M1), Ochratoxin A, Patulin, Fumonisine (B1 and B2), Zearalenone, Deoxynivalenol, soon T-2 und HT-2 toxin and ergot. Avrupa Birliğinde düzenlemeler: Aflatoksin (B1, B2, G1, G2 ve M1), Okratoksin A, Patulin, Fumonisin (B1 ve B2), Zearalenon, DON, ve yakında T-2 ile HT2 toksin ve ergot. Legislation as result for occurrence data Oluşma verilerinin sonucu olarak mevzuat 8 8 Regulated in Japan: Citrinin ( red rice - Angkak) Japonya daki düznlemeler: Sitrinin ( Kırmızı pirinç - Angkak) Regulated in Slovakia: Sterigmatocystin Slovakya daki düzenlemeler: Sterigmatocystin Less data available but occasionally reported to be found: Ennianine (Beauvericin), Cyclopiazonsäure, 4

Regulated in the EU: Aflatoxin (B1, B2, G1, G2 and M1), Ochratoxin A, Patulin, Fumonisine (B1 and B2), Zearalenone, Deoxynivalenol, soon T-2 und HT-2 toxin and ergot.

5 Regulation 1881/ Aflatoxins: Groundnuts, dried fruits, cereals (incl. maize), spices (chilli, pepper, ginger), milk. Aflatoksinler: Yer fıstığı, kuru meyveler, tahıllar (mısır dahil), baharatlar (pul biber, kara biber, zencefil), süt. Ochratoxin A: Cereals (inkl. maize), dried vine fruits, grape juice, gcoffee, wine. Okratoksin A: Tahıllar (mısır dahil), kuru üzüm, üzüm suyu, kahve, şarap Patulin: apple juice and apple juice products. Patulin: elma suyu ve elma suyu ürünleri DON/ZON/T-2&HT-2: cereals and products thereof. DON/ZON/T-2&HT-2: tahıl ve tahıl ürünleri RASFF as source for occurrence Kaynak olarak RASFF

maize), dried vine fruits, grape juice, gcoffee, wine.

6 National Data (The Netherlands) Ulusal Veriler (Hollanda) Source: M Spanjer Scientific publications as source Kaynak olarak bilimsel yayımlar Single studies give often a good snap-shot of the status quo, but might need interpretation in some cases: Tekli çalışmalar durum hakkında genellikle iyi görüntü vermekle beraber bazı durumlarda yoruma ihtiyaç duyulabilir: Way of reporting of results as pos. or neg. Sonuçların pozitif veya negatif olarak raporlanma şekli Number of samples tested Test edilen örnek sayısı Method used for analysis Analiz için kullanılan metot 6

hakkında genellikle iyi görüntü vermekle beraber bazı durumlarda yoruma ihtiyaç duyulabilir: Way of reporting of results as pos. or neg.

7 Scientific publications as source Kaynak olarak bilimsel yayımlar Easier to use (but eventually not up to date) to observe trends are reviews on occurrence data Oluşum verilerinin değerlendirilmesini kolaylaştırır. Overview over longer period (e.g. trends) Daha uzun zamanlı gözlem Comparison of data Verilerin karşılaştırılması Comparison of conclusions!!! (e.g. processed vs. raw organic products) Sonuçların karşılaştırılması (örn. işlenmişe karşı organik ürünler Examples of published Surveys Abstract (2003) An indirect competitive double antibody plate-elisa was employed for detection of aflatoxin B1 in naturally contaminated poultry feed ingredients. The feed samples which were found negative by chemical assay employing thin layer chromatography were tested by plate-elisa. Out of samples tested, 23 samples (85.18%) were found positive... Abstract (2001) The present investigation was undertaken for analysis of aflatoxin B1 in naturally contaminated poultry feed/ingredients using an indirect competitive double antibody dot-elisa 92.21% (71) samples of poultry feed/ingredients were found positive for aflatoxin B1 which include 21 samples found negative by pressure mini column and thin layer chromatography. 7

raw organic products) Sonuçların karşılaştırılması (örn.

8 Examples of published Surveys Abstract (2006) OTA and DON were assessed by HPLC in organically and conventionally produced beers sold in Belgium High OTA contamination above the limit of 200 ng/l (up to 1134 ngl-1) occasionally occurred in organically produced beers. A complementary survey performed with the same brands in 2005 did not confirm this accidental presence of excessive OTA loads The overall incidence of DON was 67 and 80% in conventional and organic beers, respectively. Abstract (2006) It also aimed to survey the presence and amounts of DON, OTA and ZEN in Belgian conventionally and organically produced wheat grain and in wholemeal wheat flours Organic and conventional wheat samples harvested in 2002 and 2003 in Belgium were analysed for DON, OTA and ZEN, while wholemeal wheat flour samples were taken from Belgian retail shops and analysed for OTA and DON. Conventional wheat tended to be more frequently contaminated with DON and ZEN than organic samples, Wheat samples collected in 2003 were less affected by DON and ZEN than the 2002 harvest. Organic wholemeal wheat flours were more frequently contaminated by OTA than conventional samples The opposite pattern was shown for DON, organic samples being more frequently contaminated than conventional flours... SCOOP Data

A complementary survey performed with the same brands in 2005 did not confirm this accidental presence of excessive OTA loads The overall incidence of DON was 67 and 80% in conventional and organic

9 Changes over time Zaman içinde değişim Mycotoxin occurrence is irregular over time: Mikotoksin oluşumu zamandan bağımısızdır but a known problem over the centuries (ergot in Europe) fakat yüzyıllardır süren problemler bilinmektedir (Avrupa da ergot) With the invention of removal practices (so called trieur ), they could be removed, until it was observed that the sclerotia of ergot changed size in some regions (due to dryness?) and observed changes can be extreme (e.g. aflatoxins in Kenya) Saptanan değişimler ekstrem olabilir (örn. Kenya da aflatoksin) Changes over time Zaman içinde değişim Irregular waves of aflatoxin occurrence for not clearly identified reasons (wrong storage, bad or little harvest) caused death of humans especially in outbreaks during 2004 and yılları arasında insan ölümlerine yol açan aflatoksin oluşumundaki dalgalanmaların sebepleri belirlenememiştir. Occurrence data is likely to change over time!!! Oluşum verileri zaman içinde değişim göstermektedir!!! 9

changed size in some regions (due to dryness?) and observed changes can be extreme (e.g. aflatoxins in Kenya) Saptanan değişimler ekstrem olabilir (örn.

10 Pre harvest occurrence Hasat öncesi oluşum Depending on the commodity the occurrence can be localised or more homogeneous in the material Oluşum ürünün türüne göre heterojen ya da homojen olabilr. Peanuts: 1/3000 to 1/ Post harvest occurrence Hasat sonrası oluşum Lot ( 19 ppb) Uncertainty 10

türüne göre heterojen ya da homojen olabilr.

11 Visible and hidden mycotoxins Görünür ve gizli mikotoksinler Other forms of known mycotoxins that are relevant for the interpretation of occurrence data (if knowledge is available!!!) Bilinen mikotoksinlerin diğer formları So called. hidden mycotoxins Gizli mikotoksinler Fumonisine (P. Scott, U. Meister, H.U. Humpf) Zearalenone ZON-glucoside (Schneeweiss, et.al. 2002) Deoxynivalenol DON-glucoside (Berthiller et.al. 2005) Mycotoxins in animal products Hayvansal ürünlerde mikotoksinler Aflatoxin M1 metabolite of Aflatoxin B1 (found in milk (suffix M). The transfer ratio is usually 1 3 % AfB1 AfM1 Aflatoksin B1 in metabolitidir (Sütte bulunur) Transfer oranı genelde %1-3 arasıdır. Ochratoxin A has a rather long half time (t½ or t0.5) of ~16 days in some animal and human tissues (e.g. blood and liver) due to its binding affinity to certain proteins. İnsanların ve bazı hayvanların dokularında yarılanma süresi belirli proteinlere bağlanma yetisi sebebi ile biraz daha uzundur. 11

al. 2005) Mycotoxins in animal products Hayvansal ürünlerde mikotoksinler 21 22 Aflatoxin M1 metabolite of Aflatoxin B1 (found in milk (suffix M).

12 Some useful links FAO HACCP Manual for Mycotoxin Prevention ftp://ftp.fao.org/docrep/fao/005/y1390e/y1390e00.pdf Code of Practise for the Prevention and Reduction of Mycotoxins Activity and Institute Information IRMM General Info Job opportunities at IRMM CRL mycotoxin activities 12

pdf Activity and Institute Information 23 24 IRMM General Info http://www.irmm.jrc.

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