1. 1 WP 5 – Fusarium and mycotoxin analysis (conclusions) P4 Eurofins Analytik GmbH (Germany) - Scarlett Biselli P6Institute for Cereal Research Bergamo (Italy) - Rita Redaelli P8Experimental Institute for Cereal Research of Fiorenzuola d´Arda (Italy) - Valeria Terzi P15Agrotest Fyto, Ltd. (CR) - Ivana Polisenska AVEQ -1st project meeting Clermont Ferrand, 19.-21.9. 2007

2. 2 WP 5 – Fusarium and mycotoxin analysis - tasks and questions 1)Which toxins are we to analyse? 2)Which Fusarium species are we to look for? 3)How to cope all of the samples - logistic? 4)Analyses; Methods AVEQ -1st project meeting Clermont Ferrand, 19.-21.9. 2007

3. 3 1)Deoxynivalenol (DON), T-2 toxin Based on the EU limits for contaminants in foodstuffs: Commission Regulation (EC) 1881/2006 DON ZEA AVEQ -1st project meeting Clermont Ferrand, 19.-21.9. 2007 F. sporotrichoides F.poae F. langsethiae T-2 + HT-2 F. graminearum F. culmorum Each sample will be analysed by ELISA for DON and T-2 toxin.

4. 4 1)Deoxynivalenol (DON), T-2 toxin AVEQ -1st project meeting Clermont Ferrand, 19.-21.9. 2007 F. graminearum F. culmorum DON; ZEA; T-2 + HT-2 cereals unprocessed1250; 100; 100 ppb oats unprocessed1750; 100; 500 ppb cereals intended for 750; 75; 200* ppb direct human consumption, cereal flour…*oat products Proposed limits

5. 5 2)Fusarium species identification AVEQ -1st project meeting Clermont Ferrand, 19.-21.9. 2007 PCR identification and quantification - P8 P8 will identificate isolates for production of inoculum (F. avenaceum, F. graminearum, F. culmorum and F. sporotrichoides) P8 should get these Fusarium colonies intended for preparing inoculum till the end 2007 from all of the WP4 partners -quantification of the amount of mycelia – standards on each location – P8 will obtain samples from WP4 (after next vegetation period)

6. 6 3)Sample selection for analyses 300 accessions - inoculated (I) + 10 (?) controls x 5 replications - I + 10 (?) controls x 2 replications - non-inoculated (NI) + 1 wheat each year x 2 (I + NI) x 3 locations(Germany, Romania, CR) ___________ 1113 samples+ 50 (?)winter types from Italy AVEQ -1st project meeting Clermont Ferrand, 19.-21.9. 2007 LC-MS/MS potential is 100 analyses/each year: - comparative study (T-2/HT-2) - verification of the results - analyses of other mycotoxins) WP5 can analyse 550 samples for 2 mycotoxins each year by ELISA - samples must be selected Italy will cultivate winter types (WT) only, NI

7. 7 3)Sample logistic for the labs AVEQ -1st project meeting Clermont Ferrand, 19.-21.9. 2007 WP5 can analyse: In GermanyIn Czech R.In ItalyIn Romania WP4 wil produce: 371 50(?) WT 371 In Czech R. – P15In Italy – P8In Italy –P6 50 250 ? ? ? ?

8. 8 3)Sample logistic for the labs AVEQ -1st project meeting Clermont Ferrand, 19.-21.9. 2007 Decreasing the number of samples: - is it necessary to analyse all standard replications ? - selection of genotypes for analyses: heavily infected genotypes should be omited (but what if no FHB symptoms on panicles in the field will be visible?) ….. To be discussed at the next meeting – after the WP4 field trials – based on their results

9. 9 4)Analyses, methods AVEQ -1st project meeting Clermont Ferrand, 19.-21.9. 2007 -harmonisation of the labs (reference samples will be used, the same ELISA kits for analyses) -non-dehulled samples for analyses according to (EC) 1881/2006 -sample homogenisation: sample preparation from incremental samples, milling more (cca 100g) than we will need for analyses (cca 20g) -necessity to keep seed for possible re-analyses or more precise analyses (HPLC) based on ELISA results - should be ensured by WP4

10. 10 AVEQ -1st project meeting Clermont Ferrand, 19.-21.9. 2007 Thank you for your attention