Highlights of the 1^st workshop

Place: Wageningen, the Netherlands, 21-22 January 1999.

• Task group 1: Reduction of Mycotoxin Contamination during Crop Production
• Task group 2: Reduction of Mycotoxin Contamination during Transport and Storage
• Task group 3: Reduction of Mycotoxin Contamination during Processing of Grain
• Task group 4: Improved Methodology to Measure Mycotoxin Contamination
• Task group 5: Integrated Chain Wide Approach

Chairmen were elected for each Task group.

Task group discussions resulted in agreements on

• the exchange of knowledge and information between the partners, especially on trichothecenes and fumonisins, measurements and regulations
• The collaboration on scientific research between partners, e.g. a ring-test to screen winter wheat varieties for their resistance to Fusarium carried out at various locations in Europe
• the exchange of trainees, e.g. to teach people from other laboratories in measuring DON concentrations in contaminated wheat samples with various techniques

Highlights of the 2^nd workshop

Place: Paris, France, 31 March - 1 April 2000.

The number of participants of the Concerted Action Mycotochain was extended with three French partners during the second workshop. These three new members are IRTAC (Institut de Recherches Technologiques Agro-alimentaires des Céréales), ITCF (Institut technologique des céréales et des fourrages) and Limagrain.

IRTAC is an independent organization and provides to be a forum for people in the cereal business. It comprises of 16 institutes and 28 private companies. IRTAC is the French Representative of the ICC (International Association for Cereal Science and Technology). ITCF (Institut technologique des céréales et des fourrages) is involved in funding both private and public research. The fields of actions are cereals, maize, potatoes and forage. Their special interest is in harvest and storage of food and non-food products. Limagrain is a breeding company.

In 1999, an incident of high levels of DON in breakfast cereals (up to 2000 µg/kg) was detected in the Netherlands. In reaction to these findings, the Dutch Government established an action limit of 1000 µg DON/kg wheat. In addition, the National Institute for Public Health and the Environment (RIVM) was asked to carry out a toxicological assessment and risk analysis for DON. In reaction to the report of the RIVM, the Dutch Government took the following actions:

The action limit was lowered to 500 µg/kg wheat product for the harvest of 1999 (in close consultation with the industry)

The Dutch Health Council was asked to prepare an opinion on DON

An agreement with the grain sector was reached that they would urgently implement their proposed measures to reduce the risk of mycotoxin contamination

• Task group 1. Reduction of Mycotoxin Contamination during Crop Production
  Ring test. Three of the partners (Ruckenbauer, De Jong and Hollins) have set up a ring test in which a set of 17 cultivars will be tested on 6 locations in 3 replicates for their resistance to Fusarium spp.
• Task group 2. Reduction of Mycotoxin Contamination during Transportation and Storage
  Task group 2 was left out because of overlap of tasks with Task groups 1 and 4.
• Task group 3. Reduction of Mycotoxin Contamination during Processing of Grain
  Processing. Two partners showed an effect of milling wheat to reduce the content of DON in the flour with respect to the original level (to about 60%). However, it resulted in the elevation of the contamination levels (up to 300% of the original wheat level) in the bran and other by-products. This may lead to problems for the use of these products, for instance for animal feeds.
• Task group 4. Improved Methodology to Measure Trichothecene and Fumonisin Contamination.
  Trichothecenes. In a study where trichothecenes were measured, it was shown that there was a large variation (25-45 %) between labs using GC methods. This is an important problem that needs to be solved before true regulations can be set. Recommendations were presented to decrease this problem.
  DON. A draft report was presented with results of a Dutch ring assay for the analysis of DON in wheat organized by TNO at the request of the Commodity Board for Arable Farming in the Netherlands. The aim of the ring assay was an inventory of the comparability of results of DON analyses generated by twelve laboratories using different methods (GC, HPLC, and ELISA). The results showed that there was a strong intra- and inter-laboratory variation in DON levels found in the assay.
  Fumonisins. The method, which was developed in the past to analyze fumonisins in maize, gave bad recoveries in the case of analyzing fumonisins in corn flour or cornflakes. It seemed that the extraction step of fumonisins from the materials and the clean-up step of the crude extracts are critical.
• Exchange of trainees.
  Several proposals were submitted for the exchange of trainees between laboratories.

Highlights of the 3^rd workshop

Place: Martina Franca, Italy, 19-20 April 2001.
The workshop was organized by ITEM, Bari, Italy.

A new member at the meeting: The University of Hohenheim - the first member of Germany.
The University of Hohenheim studies the genetic analysis of Fusarium aggressiveness, host resistance and mycotoxin production in rye and wheat. Furthermore, genetic variation within F. graminearum and F. culmorum populations is studied.

Research Activities at ITEM in Bari, Italy - Dr. Angelo Visconti
Research at ITEM focuses on the two major areas: 
1. Food safety and food quality 
2. Bio-pesticides and bio-active microbial metabolites 
The first area covers the following tasks: prevention of mycotoxicological risks in food products, evaluation of toxicological risks derived from mycotoxins in food and feedstuffs, exploitation and standardization of traditional processing of typical foods. The second area deals with antagonistic fungi and bacteria, mycoherbicides and mycopesticides. 

Fusarium mycotoxin problems in Europe 
In 1999 and in 2000 growing conditions were not very favorable for Fusarium infection and spread (wet weather conditions during the flowering period of the crop) as compared to the year 1998. In most countries hardly any seed lot was found with infection levels over 500 ppb.

 Early detection of toxigenic Fusarium species and ochratoxigenic fungi in plant products

• The development of molecular diagnostic methods (biochips, quantitative PCR, NASBA) for early detection of toxigenic Fusarium species and ochratoxigenic fungi in plant products;
• New and more sensitive immunological tests for detection of mycotoxins in foods;
• Biochemical and molecular studies to characterize genes responsible for ochratoxin A (OTA) synthesis;
• The development and validation of mycotoxin prevention strategies and the implementation of the HACCP-approach for OTA and Fusarium toxins.

 European Mycotoxin Awareness Network (EMAN)

• To build up data-basis on literature references and individual contacts
• To establish national contacts throughout Europe countries
• To create and maintain up-dating of a web-site called http://www.mycotoxin.org
• To publish newsletters and fact/information sheets on general or specific topics
• To run on-line training sessions together with internet forum

 Safemaize

• To develop improved maize genotypes with increased resistance to  F. verticillioides.
• To characterize F. verticillioides isolates from Europe and Africa.
• To develop screening methods to evaluate maize genotypes for resistance to the fungus.
• To evaluate the synergistic effect of plant anti-fungal defense genes against  F. verticillioides.
• To produce transgenic maize with stable inheritance of selected plant anti-fungal genes, and to evaluate resistance in the field.
• Risk assessment of fungal bio-control agents
• Sustainability, product safety and quality in cereals: development of novel quantitative models for risk assessment for mycotoxigenic  Fusarium species

Task group 1. Reduction of mycotoxin contamination during crop production.
Preliminary results on the ring test
A ring test was set up in which 17 cultivars were tested on 6 locations in 3 replicates for their resistance to Fusarium spp. The most resistant cultivar with an overall good response is 'Petrus'. Preliminary conclusions that could be drawn are: 1. Within this sample of European cultivars great differences exist in resistance against Fusarium head blight; 2. The genotypes x environment interactions are significant and show the urgent necessity of multi-environment testing; 3. To compare results between breeders the scoring should be better defined and training in scoring would be helpful.

Task group 3. Reduction of mycotoxin contamination during Processing of grain
Developments in the risk analysis process regarding mycotoxins in the framework of the Codex Committee on Food Additives and Contaminants
The Codex Committee on Food Additives and Contaminants (CCFAC) is one of the horizontal Committees operating under the Codex Alimentarius Commission (CAC). The CAC is an international organization supported by FAO and WHO, aiming at facilitating world trade and protecting the health of the consumer by developing international standards for foods and feeds. The CCFAC develops standards in a procedure which follows the principles of risk analysis as far as possible, according to rules and methods as they are laid down in the general Codex Manual and more specifically in the Codex General Standard for Contaminants and Toxins in Food. Reports of Committee meetings and of JECFA (WHO and FAO Committee of experts, which acts as a scientific advisory committee to the CCFAC) are available on the Codex website http://www.codexalimentarius.net

Task group 4. Improved methodology to Measure trichothecene and fumonisin contamination
ASTER
The ASTER system is developed by TNO Nutrition and Food Research and can be used to sort sticky material like compost or wood chips. Additionally it is able to clean and grade materials like grains, seeds, beans, herbs and sugar. It is based on the principle that aerodynamic forces are influenced by a particle's velocity, size and shape. An exploratory investigation has shown that ASTER can be used to upgrade DON-contaminated wheat. In a first experiment DON contamination could be reduced with about 40 %.

1. Conduct a hazard analysis
2. Determine critical control points
3. Establish critical limits
4. Establish a monitoring system
5. Establish corrective actions
6. Establish verification procedure
7. Establish documentation system

Monitoring is the continuous or scheduled measurement or observation at a Critical Control Point (CCP) to assess whether the step is under control, i.e. within the specified critical limits.