WP 5 – Fusarium and mycotoxin analysis

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WP 5 – Fusarium and mycotoxin analysis

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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- what can we offer?

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P4 (Scarlett) - mycotoxin analyses LC-MS/MS (multitoxin method, DON, T-2, HT-2, zearalenone, diacetoxyscirpenol, 3- and 15- acetyldeoxynivalenol) - 200 samples

P6, P8, P15 (Rita, Valeria, Ivana) - ELISA mycotoxin analyses, DON, T-2, ZEA - 2200 analyses

P8 (Valeria) - Identification of Fusarium species - 200

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- tasks and questions

1) Which toxins are we to analyse?

2) Which Fusarium species are we to look for?

3) How to manage huge amount of samples?

4) Analyses; Methods

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AVEQ = Avena genetic resources for quality in human consumption

EU legislation setting maximum levels for certain contaminants in foodstuffs: Commission Regulation (EC) 1881/2006

DON (deoxynivalenol), ZEA, T-2 + HT-2

New discussed EU limits for T-2 and HT-2(to be fixed in 200?)

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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

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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.

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2) Which Fusarium species?A

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What should be the purpose ...

A) to analyse naturally infected oats to determine Fusarium species and their mutual proportions (variability in years, locations)

B) to analyse inoculated oats- to confirm that (only) the Fusarium species which was used for inoculation is present ?- quantification of the amount of Fusarium mycelium for studying relationships between mycotoxins content ?

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A) to analyse naturally infected oats- which species?

Tekauz and Mueller (2006) - in Canada F. graminearum, F. poae, F. sporotrichoides and F. avenaceum

Parrika et al. (2006) - in Finland F. langsethiae, F. poae, F. culmorum, F. graminearum, F. avenaceum

Imathiu et al. (2006) - in GB F. langsethiae

CR - 2006, 9 oat samples, 16 Fusarium isolates

7x F. sporotrichoides

6x F. poae

2x F. culmorum

?F. langsethiae?

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F. langsethiae

- newly described Fusarium species- similar to F. poae and F. sporotrichoides- produces T-2 and HT-2 toxins- isolated from all types of cereals, more common in oats- found in Sweden, Norway, Russia, Austria, Finland- hard artificial infection at flowering

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3) Sample selection for analyses

Inoculated plots:300 accessions + 10 controls each year x 4 locations (Germany, Italy, Romania, CR)______________1240 samples

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In case of 2 (DON, T-2):

550 samples ELISA + 100 HPLC/per year

ELISA potential: 1100 analyses/per year (?number of mycotoxins analysed?)

LC-MS/MS potential: 100 analyses/each year

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3) Sample selection for analyses

LC-MS/MS :- detection limit of 10 ppb- HT-2 toxin- more toxins in one analysis - expensive

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Discussion about LC-MS/MS role

- verification of ELISA results, interesting samples (low visible infection, low ELISA levels…)

- analyses of other mycotoxins

- comparative study - relationship between T-2 and HT-2 (naturally infected, inoculated)

ELISA :- detection limit of 5-50 ppb- HT-2 toxin- one toxin in one analysis- reasonable costs

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4) Analyses, methods A

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- harmonisation of the labs (1-2 reference samples, interlaboratory comparison)

- the same kits for analyses ?

- sample amount per analysis ?

- logistic of sample distribution ?

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Year 2007

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Year 2008

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Year 2009

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Year 2010

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WP 5 - Time table

Documents

WP 5 – Fusarium and mycotoxin analysis