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WP 5 – Fusarium and mycotoxin analysis

(conclusions)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

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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

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1) Deoxynivalenol (DON), T-2 toxin

Based on the EU limits for contaminants in foodstuffs:

Commission Regulation (EC) 1881/2006

DONZEA

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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.

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1) Deoxynivalenol (DON), T-2 toxinA

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F. graminearum

F. culmorum

DON; ZEA; T-2 + HT-2

cereals unprocessed 1250; 100; 100 ppb

oats unprocessed 1750; 100; 500 ppb

cereals intended for 750; 75; 200* ppbdirect human consumption, cereal flour…*oat products

Proposed limits

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2) Fusarium species identificationA

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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)

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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

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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

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3) Sample logistic for the labsA

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WP5 can analyse:

In Germany In Czech R.In Italy In Romania

WP4 wil produce:

371 50(?) WT 371371

In Czech R. – P15In Italy – P8In Italy –P6

50 250250

?? ? ?

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3) Sample logistic for the labsA

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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

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4) Analyses, methods A

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-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

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Thank you for your attention