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Development, validation and harmonisation of screening and confirmatory tests to distinguish zeranol abuse from fusarium toxin contamination in food animals

Contract nr: FAIR-CT97-3443
Project nr: 3443
Project type: SC
Starting date: 01/02/1998
Duration: 36 months
Total cost: 1,045,100 EUR
EC Contribution: 850,000 EUR
Scientific Officer: Michel VAN DEN BOSSCHE
Research topic: Technological instruments and methods
Acronym: Natural zeranol

Zeranol is a non-steroidal oestrogenic growth promoter. It increases liveweight gain in food animals following implantation. The use of zeranol for growth promotion in food animals was banned in the EU in 1985. All Member States have been required, since 1986, to establish National Surveillance Schemes to monitor animals for possible abuse of banned substances, including zeranol. The ability of Member States to implement the terms of Council Directive 96/23/EEC has been compromised by recent reports from New Zealand, and elsewhere, showing that zeranol occurs naturally in urine and bile taken from cattle and sheep. The New Zealand study suggested that naturally occurring Fusarium spp. toxins could be metabolised to zeranol. Enforcement of the ban on zeranol has been further compromised by similar reports of naturally occurring zeranol from several National and Community Reference Laboratories in the EU. These findings prevent the smooth implementation of Council Directive 96/23/EEC because:
1. All current immunochemical screening tests exhibit cross-reactivity to a greater or lesser extent with zearalenone, a-zearalenol and ß-zearalenol - the three principal toxins formed by the Fusarium spp. fungi. These screening assays will therefore show "positive" results in all samples containing these toxins, irrespective of whether zeranol itself is actually present or not.
2. Confirmatory analysis will prove whether zeranol is present or not. But, GC-MS will not prove whether it is natural in origin.
3. There are no criteria to differentiate zeranol abuse from contamination in meat from 3rd country imports into the EU.

The increasing financial penalties being imposed on offending producers throughout the EU as a result of the implementation of Council Directive 96/23/EEC makes it essential that a proper decision is reached by competent national authorities.

The central objective of this project is to improve reagents and procedures for the technical implementation of Community policy on the prohibition of the use of zeranol as an anabolic agent in food animal production.

This project directly addresses Objective 4.22 of the workprogramme.
It will answer the three problems posed above by:
1. The development of rapid, specific immunochemical screening tests that will specifically and separately detect and quantify zeranol and a-zearalenol in a sample;
2. Identifying those food species that are capable of metabolising Fusarium spp. toxins to zeranol;
3. The provision of quantitative criteria to differentiate between zeranol abuse and natural environmental contamination in susceptible species by measurement of concentrations of zeranol and the Fusarium spp. toxins in body fluids; and
4. The provision of quantitative criteria to differentiate between zeranol abuse and natural environmental contamination with zeranol by measurement of concentrations of zeranol and the Fusarium spp. toxins in tissues, facilitating monitoring of 3rd country imports into the EU.

Current situation/results:
Dry chemistry time-resolved fluorescence immunoassays (TRFIAs) for zeranol and a-zearalenol have now been developed and validated. These have both the specificity and sensitivity required by this project. The assays have been transferred to three other laboratories within the consortium. These laboratories have performed their own validation on the assays and have conducted a blind Ring Test of urine samples spiked with combinations of zeranol, a-zearalenol or Fusarium spp. toxins.
Chemical methods for the detection of zeranol and the Fusarium spp. toxins have been developed. These will be used to measure the concentration of zeranol and the Fusarium spp. toxins present in all samples that screen positive, using the TRFIAs, in a survey of animal urine samples due to start shortly in four Member States. Parallel studies measuring the stability of zeranol and the Fusarium spp. toxins in a range of biological matrices are nearing completion.


Glenn Kennedy
Veterinary Sciences Division
Department of Agriculture and Rural Development
Tel: +44 2890 52 56 51
Fax: +44 2890 52 56 26

  • Vasilis VOZIKIS
    National Agricultural Research Foundation
    26th October Street 80
    Tel: +30 31 55 20 24
    Fax: +30 31 55 40 22

  • Leendert A. VAN GINKEL
    National Institute of Public Health and the Environment
    Antonie Van Leeuwenhoeklaan 9
    NL-3721 Bilthoven 1
    Tel: +31 30 274 27 46
    Fax: +31 30 274 44 03

  • Luisa Maria RIBEIRO
    Laboratorio Nacional de Investigacao Veterinaria
    Estrada De Benfica 701
    P-1500 Lisboa
    Tel: +351 21 711 53 30
    Fax: +351 21 711 53 84

    The National Swedish Food Administration
    S-751 26 Uppsala
    Tel: +46 18 17 56 12
    Fax: +46 18 10 58 48

  • Timo LÖVGREN
    University of Turku
    Department of Biotechnology
    Biocity 6A
    FIN-20520 Turku
    Tel: +358 2 333 80 51
    Fax: +358 2 333 80 50

  • Antti IITIA
    InnoTrac Diagnostics Oy
    Biocity 7A
    FIN-20520 Turku
    Tel: +358 2 333 80 87
    Fax: +358 2 333 80 50
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