Novel integrated strategies for worldwide mycotoxin reduction in food and feed chains

2009 - 2013

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Results

Project overview at 36 months

Pre-harvest

  • 80-90% reduction in DON contamination was reached by using fungicide optimization technology;
  • additional 50 % reduction compared to the best technology found until now was reached by developing a better nozzle composition;
  • Examination of QTLs in wheat/maize for identification of resistant traits;
  • Microarray methodology was developed for examining host/pathogen “maize vs F. verticillioides” interaction and 800 genes were identified in the host/pathogen interaction;
  • Reduction in aflatoxin contamination peanuts (up to 97%) was reached by using specific antagonists in Argentina;
  • Fusarium spp. in stubble of maize and FHB and DON in wheat caused by F. graminearum was reduced by using antagonists decreasing of about 70%;
  • Economics of large scale production and commercialisation of 4 BCA has been completed;
  • Field trials in the Netherlands, Italy, Nigeria and Argentina have been done with selected BCA in wheat, maize, peanuts;
  • Up to 79% AFs control reduction in maize was achieved with non-toxigenic strains in Nigeria;
  • Standardised data systems in place for: cropping systems, locations, meteorology, analytical data for model validation and development of risk maps;
  • DON in wheat data collected in MycoRed has been used for validation of an existing model;
  • A database has been completed prepared containing all available and relevant information for modelling development for F. verticillioides and FUM, A. flavus and AFs both in maize, and for OTA in grapes.

Post-harvest and processing

  • The relationships between environmental factors and dry matter loss relevant to EU legislative limits (DON, FUM and AFs) have been identified in wheat, maize and hazelnuts;
  • Examination in vitro of O3 effect on germination and growth on relevant mycotoxin producing fungi;
  • Novel compounds potentially useful for post-harvest control of DON, FUM and AFs in these food chains have been identified;
  • A system based on wireless sensor devices for temperature, humidity and CO2 monitoring has been developed and completed for introduction into pilot scale grain silos;
  • Agricultural by products and commercial products proved to be able in vitro for the ability to bind simultaneously aflatoxins B1, ZEA, FB1, OTA and DON;
  • Some yeast strains were able to detoxify aflatoxins B1 and OTA up to 48% and 93% respectively;
  • Sulphuration dehulling/peeling and sorting of apricot seed reduced aflatoxins B1 significantly (75- 91%);
  • 50% reduction of aflatoxin content in roasted almonds was achieved.


Advanced technologies for diagnostics and novel approaches to control mycotoxigenic fungi

a) Molecular methods

  • Studies on diversity of Aspergillus, Fusarium and Penicillium (1150) from several host plants have been carried out on a large collection of fungal strains using molecular tools and toxin profiling;
  • Primers pair CLF2/CLR2 have enabled to successfully discriminate 3 groups of Aspergillus species on grapes;
  • TaqMan quantitative PCR assay for tricothecenes type A species (F. langhsetiae and F. sporotrichioides) has been developed;
  • PCR system for P. verrucosum was developed to follow the growth in wheat;
  • The effect of temperature and water stress on AFs regulatory genes was studied on almond based matrices;
  • Light at different wave length appears to be a promising method to control growth and toxin production.
b) Analytical methods
  • LC-MS and LC-HR-MS based methods have been developed and validated for multi-mycotoxin analyses in a range of food matrices from the chosen food chains;
  • Fungal metabolite profiling of cultures was carried out using a database of >800 metabolites;
  • A sensitive LC-MS/MS method was developed for simultaneous determination of multi-biomarkers for OTA, AFA B1, ZEA, FB1 and DON in human and animal urine;
  • Over 500 samples obtained from other Partners and WPs have been analysed for 250+ metabolites as a horizontal activity;
  • Rapid test kits (strip tests) for the detection of DON, AFA and FUMs have been thoroughly validated and checked for cross-reactivity against conjugated and other altered forms of mycotoxins.
  • Masked DON- and ZON-species contributed to the overall mytoxin burden as their concentration equals 10-30% of the concentrations of their parent toxins.


Project’s overview at 24 Months

  • General appreciation for MycoRed research, approach and initiatives at global level, growing interest towards the project and our community, consolidation of scientific position in the international scenario (17 scientific alliances)
  • Positive and encouraging scientific results, including 20 scientific publications on international reviews arising from WPs and the “Mycotoxin Reduction in Grain Chains: A Practical Guide“, edited by J.F. Leslie and A. Logrieco, co-written by 11 MycoRed Beneficiaries scientists
  • Good supporting and participation by partners and network in preparing joint initiatives also abroad (i.e. Conferences in Malaysia, South Africa – 60 project’s presentations in the framework of international conferences and seminars)
  • Continuous interaction among Beneficiaries, WP leaders and Coordinator; strong coordination and management efforts
  • 24 Months Deliverables: 33 Deliverables obtained and reports submitted (2 postponed to 36 Months);
  • 24 Months Milestones: 42 milestones achieved (reports or collections, platforms);
  • 18 Months report and financial statements presented and submitted by Coordinator in NEF system
  • Mid-Term review successful carried in Brussels on September 1st, 2011.

Interactions with other EU projects


EU MoniQa project co-organizer of the training course in Malaysia: “Training course for capacity building in mycotoxin-safe food trade”, 2010 .
A Young Ph. Student from MoniQa cooperated with CNR on a topic of MycoRed interest too, sharing knowledge and methodologies.

A lesson and practical training in laboratory on multiplex dipstick developed within the EU project CONffIDENCE held at training course "Detection techniques for mycotoxin and toxigenic fungi in the food chain” in Bari, IT, 2010 .


  • Presentation of MycoRed project at the EU funded Biocircle project International Conference "European Agricultural Policies – Going Global. How does Europe integrate global needs in its agricultural strategies, policies and instruments?“, in March 2010, Rome, Italy.
  • Participation at international SELAMAT Workshop on “Pesticide Residues & Mycotoxin and Food Safety” in Beijing, China, in September 2010, with the main lecture by Coordinator about the MycoRed initiatives for worldwide mycotoxin reduction in food and feed.
  • Presentation of MycoRed success story within the EU funded Bio-Net  project, that aims to provide high quality support for organizations participating in FP7 projects. A video focusing on MycoRed as case story will be realized in next fall 2011.
  • Participation at workshop: “Thematic priority setting for EU, Caribbean & Central America in research and innovation”, in March 2011, Santo Domingo, Dominican Republic, in the framework of “ ENLACE – EUCARINET” programme by MycoRed Coordinator as invited European expert on Food Safety and MycoRed Home education session held at Universidad Nacional Evangelica.


Progresses at 18 months


WP1
 In wheat 40 cvs were compared with three different inoculation methods in two location against four isolates of Fusarium. Visual symptoms, kernel infection and DON contamination was measured. Two populations were searched for QTLs in Szeged, the Hobbit sib series in JIC. For gene expression against F. verticillioides in maize three experiments were made with gene expression test by microarray methodology. In wheat farm scal and small plot fungicide tests were made with new nozzle combination on three cvs and eight fungicides. Here also the translocations of fungicide active ingredients were followed. In maize again 40 hybrids were tested in 2010 (60 ib 2009 as preliminary trial) by two inoculation methods. The field work was finished in November this year.

WP2  Selected antagonists were tested in field experiments in wheat and maize in The Netherlands, Italy and Argentina to reduce mycotoxins produced by Fusarium spp.. Various experiments were carried out in peanuts and maize aiming at the reduction of aflatoxin produced by Aspergillus spp. Economics of large scale production of selected antagonist has been preliminarily assessed.

WP3 All available data were collected to chose/develop predictive models for mycotoxin contamination in wheat, maize and grapes. A sampling protocol was developed for all the crops sampling points were defined in all countries involved; relevant mycotoxins were quantified in all the wheat samples at harvest. A questionnaire was prepared suitable to collect all data regarding geographic coordinates and cropping system of the selected crops and filled for each sample. A meteorological station reasonably close was selected for each sampling point and meteorological data were collected. All data were stored in a data base and they were/will be used for model validation.

WP4 Four aspects of the post-harvest strategies were developed during this period. The effect of environmental factors on respiration of wheat maize, and hazelnuts inoculated with key mycotoxigenic fungi were done. Data was used to develop models to relate environmental factors to dry matter loss (DML) and to mycotoxin contamination. O3 has been tested in vitro for efficacy against A.flavus. A large number of novel compounds have been screened for efficacy against F.graminearum, F.verticillioides, A.flavus, A.carbonarius, P.verrucosum and A.westerdijkae and control of mycotoxins. The best ones have now been chosen for in situ storage experiments. A sensor “ball” system which incorporates temperature, relative humidity and CO2 has been developed, and the associated hardware completed. 

WP5 Agricultural by products and commercial products have been tested in vitro for the ability to bind simultaneously aflatoxins B1, ZEA, FB1, OTA and DON. Pilot experiments were performed to determine in vivo parameters that should be use to determine the toxicity of mycotoxin and the subsequent effect of agricultural and commercial products. The ability of yeast strains to detoxify aflatoxins B1 and OTA has been initiated. The efficacy and safety of food processing procedures in reducing mycotoxin content has been initiated. We observed that sulphuration dehulling/peeling and sorting significantly reduced the content of aflatoxins B1 of apricot seed reduced aflatoxins B1. Pilot, in vitro tests were performed to asses the cellular toxicity of FB1.

WP6   The strain collection kept at CNR has been extensively increased with respect to toxigenic Fusaria, Aspergilli and Penicillia. For important species of that collection specific primers and new detection methods have been established. Molecular characterization of Aspergillus species section Nigri isolated from grapes have been performed. The influence of external stress factors on aflatoxin producing Aspergilli and on ochratoxin producing Penicillia have been analysed. Especially the influence of light of different wave length on growth of several food borne fungi and especially on ochratoxin A biosynthesis has been carried out.

WP7  The following analytical tools have been developed and/or validated: 1) LC-MS/MS or LC-HRMS based methods for a) multi-mycotoxin analysis in food matrices; b) fungal metabolite profiling in cultures; c) masked DON- and ZON species in food d) detection of mycotoxin biomarkers in urine and e) determination of novel biomarkers as well as 2) strip tests for rapid test systems.

WP8 MycoRed different dissemination actions have been taken to introduce the project in the international scenario opening the road to further events, by presentations of project worldwide at international conferences and workshop, project’s website and promotional materials. International conferences, workshops and training activities, including Short Term Visits have been organized as well as networking actions at different levels.

 



Focus on activities at 15 Months

The main activities of MycoRed project are on going starting from April 2009.
Here you find news regarding the progresses on going in different Work Packages, as described by the WP leaders and the Deliverables tables per year.


Project Deliverables

Table of codes
Nature of Deliverables:
R = Report, P = Prototype, D = Demonstrator, O = Other
Dissemination level:
PU = Public
PP = Restricted to other programme participants (including the Commission Services)
RE = Restricted to a group specified by the consortium (including the Commission Services)
CO = Confidential, only for members of the consortium (including the Commission Services)





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