WP 5 – Fusarium and mycotoxin analysis (conclusions)

1. WP 5 – Fusarium and mycotoxin analysis (conclusions) AVEQ -1st project meeting Clermont Ferrand, 19.-21.9. 2007 P4 Eurofins Analytik GmbH (Germany) - Scarlett Biselli P6 Institute for Cereal Research Bergamo (Italy) - Rita Redaelli P8 Experimental Institute for Cereal Research of Fiorenzuola d´Arda (Italy) - Valeria Terzi P15 Agrotest Fyto, Ltd. (CR) - Ivana Polisenska

2. WP 5 – Fusarium and mycotoxin analysis - tasks and questions AVEQ -1st project meeting Clermont Ferrand, 19.-21.9. 2007 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

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

4. 1) Deoxynivalenol (DON), T-2 toxin AVEQ -1st project meeting Clermont Ferrand, 19.-21.9. 2007 Proposed limits DON; ZEA; T-2 + HT-2 cereals unprocessed 1250; 100; 100 ppb oats unprocessed 1750; 100; 500 ppb cereals intended for 750; 75; 200* ppb direct human consumption, cereal flour…*oat products F. graminearum F. culmorum

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. 3) Sample selection for analyses AVEQ -1st project meeting Clermont Ferrand, 19.-21.9. 2007 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 Italy will cultivate winter types (WT) only, NI WP5 can analyse 550 samples for 2 mycotoxins each year by ELISA - samples must be selected LC-MS/MS potential is 100 analyses/each year: - comparative study (T-2/HT-2) - verification of the results - analyses of other mycotoxins)

7. 3) Sample logistic for the labs AVEQ -1st project meeting Clermont Ferrand, 19.-21.9. 2007 WP4 wil produce: In Germany In Italy In Romania In Czech R. 50(?) WT 371 371 371 WP5 can analyse: In Italy –P6 In Italy – P8 In Czech R. – P15 250 50 250 ? ? ? ?

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