Agricultural research Projects adopted following the FAIR specific call for proposals on TSEs

The rist of research that was accepted in 1997

• Analysis of molecular factors affecting variability in BSE and scrapie susceptibility
• Relationship between conformation of PrP, infectivity and pathogenicity of bovine spongiform encephalopathy (BSE) as a basis for diagnosis
• Generation of bovine and ovine PrP transgenic mice for the development of improved bioassays for BSE and scrapie agent detection
• New approaches to the diagnosis and control of transmissible spongiform encephalopathies
• Measures to reduce contamination of meat and environment with CNS tissue during slaughter and processing of cattle and sheep
• Improving prospects for scrapie control in sheep and goats by study of host genotypes, TSE isolates and their in vivo and in vitro interaction
• Development of a novel diagnostic to assist quality assurance procedures in European meat production
• Separation, identification and characterization of the normal and abnormal isoforms of prion protein from normal and experimentally infected fish 

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PL973311 : Analysis of molecular factors affecting variability in BSE and scrapie susceptibility

• Coordinator: J. Williams, Roslin Institute (Edinburgh), Division of Molecular Biology (UK) Participants :
• * L. Ferretti, Consiglio Nazionale delle Ricerche, Istituto per la Difesa e la Valorizzazione del Germoplasma Animale (IT)
• * I. Olsaker, Norwegian College of Veterinary Medicine Department of Morphology, Genetics and Aquatic Biology (NO)
• * P. Kitchin, Rosgen Ltd., Limited Company registered in Scotland (UK)
This project will examine the genetic factors involved in the susceptibility of sheep to scrapie and whether there are genetic factors involved in the susceptibility of cattle to BSE. Much work has been carried out in sheep on the relationship between particular forms of the PrP gene and prevalence of natural scrapie, with particular forms in defined breeds being associated with high disease incidence (Hoinville et al Vet Rec 136 312). Studies so far have focused on variations in regions of the gene which code for the protein , and in particular on a specific part of the gene - the octapeptide repeat region. No information is available regarding the way the different forms of the gene affect susceptibility to disease or on regions of the gene which regulate its'expression. In addition although breeding experiments have pointed to PrP as the major factor governing progression of scrapie in sheep, there has been no work to formally exclude the involvement of other genes.

 In cattle despite the seriousness and extent of the BSE epidemic in the UK, little work has been done to examine the genetics of infection and progression of the disease. The coding region of PrP has been sequenced twice and three polymorphisms have been found (Goldmann et al 1991 J. Gen. Virol. 72 201). The study of the effect of the different forms of PrP on BSE susceptibility has proved inconclusive (Neiburgs et al Anim Genet 25 313) while examination of incidence of disease in calves of BSE affected and unaffected cows, is suggestive of a genetic variation in susceptibility to disease (Hoinville et al 1995 Vet. Rec. 136 312). The work that will be undertaken in this project will add to the knowledge of genetic factors involved in the development of scrapie in sheep and will test whether there are genetic factors affecting the susceptibility of cattle to BSE. The study will focus at several levels:

• 1. The complete PrP gene including flanking regulatory regions will be sequenced from both sheep and cattle. This information will be used to identify the majority of polymorphisms in the gene for both species. The frequencies of the polymorphisms found will be measured accross different breeds and populations. Selected polymorphisms, focusing on regulatory and conserved elements of the gene, will then be tested for a role in the development of disease in sheep and cattle by linkage analysis.
• 2. The expression levels of the PrP gene in affected vs unaffected individuals will be examined to see whether the level of PrP expression is associated with pre-disposition to disease.
• 3. The chromosomal region neighborouring the PrP gene will be examined for other genes that may be involved in disease. This search will initially use of genetic markers, selected from genetic maps, which will be tested for involvement in disease by segregation analysis. In addition novel expressed sequences near to the PrP gene, which may confer differing susceptibility to disease, will be sought through exon trapping using large fragment yeast artificial chromosome (YAC) clones covering the chromosomal region containing the PrP gene. The putative role of the expressed sequences identified in development of scrapie will be tested by examining expression levels in affected vs healthy sheep.
• 4. Finally the whole genome will be examined for additional loci that are involved in development of BSE in cattle. Using microsatellite markers selected to cover the whole genome at roughly 20 cM intervals (1/150th of the genome) the inheritance of chromosomal regions will be tracted from selected sires into affected and unaffected progeny. Distorted segregation of markers between affected and unaffected progeny will reveal loci that may be involved in the disease. This work will complete the basic molecular information required for studying the disease.

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PL973314 : Relationship between conformation of PrP, infectivity and pathogenicity of bovine spongiform encephalopathy (BSE) as a basis for diagnosis

Coordinator : D. Riesnev, Heinrich-Heine-Universitt Dsseldorf, Institut fr Physikalische Biologie (DE)
Participants :
• * R. Jackman, Veterinary Laboratory Agency, Central Veterinary Laboratory (CVL) Addlestone (UK)
• * S. Weiss, Ludwig-Maximilians-Universitt Munchen, Institut fur Biochemie -Genzentrum der LMU Mnchen (DE)
• * D. Dormont, Commissariat  l'Energie Atomique, Direction des Sciences du Vivant Dpartement de Recherche Mdicale, Service de Neurovirologie (FR)
• * H. Kretzschmar, Georg-August-Universtitt, Institut fr Neuropathologie (DE)
• * J. Rossier, Centre National de la Recherche Scientifique, Ile de France - Secteur Paris B, Unit de Recherche Associe 2054, Neurobiologie de la Diversit Cellulaire (FR)
• * C. Reiter, Connex Gesellschaft zur Optimierung von Forschung und Entwicklung Mbh, Connex Gmbh (DE)
According to the prion-model, BSE is transmitted by a proteinaceous infectious agent (prion); the major component is a host protein, the so-called prion-protein, which is present in the cellular form, PrPc and after infection in an abnormal form PrPBSE. The abnormal isoform of PrP triggers the transformation of the cellular into the abnormal form thereby multiplying the agent. The knowledge of the basic biological, biochemical, and biophysical features of PrPBSE is regarded as a prerequisite to develop new techniques for diagnosis.

 The molecular properties of PrPc, PrPBSE and PrP-peptides of the bovine system will be determined and compared with those of scrapie and CJD to differentiate between general principals of prion diseases and particular BSE-features like the so-called BSE-strain of the prion agent. Methods from molecular biology, biochemistry and biophysics will be applied. As an alternative way of a fast and reliable diagnosis of BSE, a cell culture susceptible to BSE infection will be established by transfection of the bovine PrP. Specified antibodies and other PrP-interacting ligands will be studied systematically to differentiate between PrPc and PrPBSE. The mechanism of neurodegeneration and the appearance of early markers after BSE infection will be studied in neuronal cell cultures. With the progress of the project the partners will study the relation between BSE infection, PrP accumulation, PrP conformation, interacting ligands, appearance of early markers and develop an optimal concept for a sensitive diagnosis. 

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PL973304: Generation of bovine and ovine PrP transgenic mice for the development of improved bioassays for BSE and scrapie agent detection

Coordinator: M.H. Groschup, Federal Research for Virus Diseases of Animals , Institute for Vaccines, Federal Research Centre for Virus Diseases of Animals (DE)
Participants :
 
• * M. Dawson, Veterinary Laboratories Agency, Virology Department, Research and Development Division, Addleston (UK)
• * J.L. Vilotte, Institut National de la Recherche Agronomique - Btiment des Biotechnologies (FR)
The need for more sensitive and cost efficient bioassay models for BSE and scrapie infectivity than currently exist is evident. The aim of this project is to generate transgenic mice which highly overexpress bovine and ovine prion protein. All transgenic mice will be analyzed immunochemically with respect to PrPc expression, tissue specificity of expression and histopathologically for spontaneous neuro- and myodegeneration in aged animals. According to the prion theory similarity in the amino acid sequence of the prion proteins of a donor in comparison to the receptor species largely determines the efficiency of transmission of an infectious agent. Accordingly transgenic mice expressing ruminant prion proteins should be more susceptible to BSE or scrapie than conventional mice and therefore useful for the development of highly sensitive bioassays for agent detection. Bovine and ovine prion protein transgenic mice will therefore be test inoculated with brain homogenates of BSE and scrapie diseased animals in order to determine their susceptibility. If animals come down with disease they will be analyzed using histopathology, immunocytochemestry and immunoblot for detection of spongiform brain lesions and accumulation of pathological prion protein. Transgenic mice will eventually be challenged with BSE brain homogenates, of which the inherent infectivity has already been determined by cattle inoculation experiments. 

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PL973306 : New approaches to the diagnosis and control of transmissible spongiform encephalopathies

Coordinator: M. Rogers, University College Dublin, University Dept. of Zoology and Biotechnology Centre (IE)
Partipicants :
 
• * J.M. S!nchez-Vizca"no, Instituto Nacional de Investigaci#n y Tecnolog"a Agraria y Alimentaria, Centro de Investigaci#n en Sanidad Animal (CISA) (ES)
• * B. Adair, The Queen's University of Belfast, Department of Veterinary Sciences (UK)
• * T. Linne, Swedish University of Agricultural Sciences, Department of Veterinary Microbiology, Section of Virology - Unit of Molecular Virology (SE)
• * M. Merza, Svanova Biotech (SE)
This project takes a comprehensive approach to the development of diagnostic tests for BSE and other TSEs. Specifically, the proposal seeks:
 
• to develop diagnostic tests for BSE and other TSEs that can be used to detect animals incubating the disease but which do not display clinical symptoms both in a live (field) test and in the tissues of animal carcasses, in an abattoir test. A preclinical test for BSE will either identify PrPsc or some other disease specific marker. Initially, the best candidate marker is PrPsc though this protein is poorly immunogenic and no antibodies are available which can discriminate between the normal and disease specific isoforms. The proposal sets out novel methods that will allow the isolation of PrPsc monoclonal antibodies. Once appropiate antibodies against PrPsc or other marker proteins have been identified, assays using these reagents will be developed and optimised.
• * to develop transgenic mice expressing PrP constructs from bovine and porcine species that provide rapid methods for assessing the levels of infectious agent in animal tissues or their products. Such animals should provide sensitive and reasonably rapid tests for BSE in different tissues and fluids derived from potentially infected animals.
• * to determine the relative efficiencies of transmission of BSE to and between pigs by the oral and intracerebral routes as part of the more general investigation into TSEs. Part of this work will also utilise a transgenic mouse model expressing the porcine PrP gene analogous to the bovine model described above.
In conclusion, this project presents a comprehensive and complete approach to the development of diagnostic tests for BSE and as part of their development and validation, provides sensitive animal models for the relatively rapid testing of beef and beef products for BSE. 

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PL973301 : Measures to reduce contamination of meat and environment with CNS tissue during slaughter and processing of cattle and sheep

Coordinator : D. Harbour, Department of Clinical Veterinary Science, Division of Molecular and Cellular Biology, University of Bristol (UK)
Participants :
 
• * J.P. Frencia, Association pour le Dvelopment de l'Institut de la Viande, Clermont Ferrand(FR)
• * J. Sheridan, Teagasc, the National Food Centre, Dublin (IE)
• * D. Tinker, Silsoe Research Institute, Company Ltd, Bedford (UK)
• * M. Owen, Meat and Livestock Commission, Public Corporation, Milton Keynes (UK).
Innovative techniques to assess, directly or indirectly, the level of cross-contamination with CNS material during conventional slaughter, dressing and butchery of cattle and sheep, will be developed. For direct measurement, nucleic acid-based (RT-PCR) and protein antigen-based (ELISA) assays will be developed to detect CNS-specific mRNA and proteins respectively. Furthermore, an innovative technique to detect prion protein (PrP), based on the affinity of PrP for molecular chaperone proteins, will be developed. For indirect measurement of cross-contamination, the use of tracer dyes and Escherichia coli K12 injected into the CNS will be assessed.

 These direct and indirect assays will be used to monitor cross-contamination of carcasses, equipment and abattoir environment (including atmospheric) with CNS material during conventional slaughter, dressing and butchery of cattle and sheep, in order to identify critical practices. Based on the results of these studies, changes to conventional methods will be devised, and tested under experimental conditions to ascertain wether cross-contamination can be reduced. Concurrently, innovative equipment (particularly saws and vacuum devices) for the safe removal of CNS material will be developed and tested for their ability to reduce or eliminate cross-contamination. Practices or equipment that do decrease the risks under experimental conditions will be assessed under commercial conditions for economic and practical viability. New practices or equipment will be assessed and demonstrated to industry. A set of guidelines for avoidance of CNS cross-contamination in the meat industry will be drafted for presentation to the EC. 

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PL973305 : Improving prospects for scrapie control in sheep and goats by study of host genotypes, TSE isolates and their in vivo and in vitro interaction

Coordinator: J.M Elsen, Institut National de la Recherche Agronomique, Station d'Amlioration Gntique des Animaux, Auzeville (FR)
Participants :
 
• * M. Dawson, Veterinary Laboratories Agency, Virology Department, Research and Development Division, Addlestone (UK)
• * N. Hunter, Institute for Animal Health, BBSRC and MRC Neuropathogenesis Unit, Edinburgh (UK)
• * M.A. Smits, DLO Institute for Animal Science and Health, Lelystad (NL)
• * M. Groschup, Federal Research Centre for Virus Diseases of Animals, Institute for Vaccines at the Federal Research Centre for Virus Diseases of Animals,Tbingen(DE)
• * A. Palsdottir, Institute of Experimental Pathology, University of Iceland, Keldur (IS)
• * O. Papadopoulos, Laboratory of Microbiology and Infectious Diseases Faculty of Veterinary Medicine Aristotle University, Thessaloniki (GR)
• * J. Roche, University College Dublin, Department of Animal Husbandry and Production (IE)
• * J. Jarp, Department of Epidemiology, National Veterinary Institute,Oslo (NO)
• * T. Baron, Laboratoire de Pathologie Bovine, Centre National d'Etudes Vtrinaires et Alimentaires, Lyon (FR)
• * M.Y. Boscher, Laboratoire d'Analyses Gntiques pour les Esp ces Animales, Jouy En Josas, (FR)
• * P. Economides, Central veterinary Laboratory, Department of Veterinary Services, Government Services, Nicosia (CY)
Differences in sheep PrP amino acid 136, 154 and 171 codons are clearly associated with susceptibility to scrapie in most outbreaks and this has opened the opportunity to breed for resistance to scrapie. However, scrapie has been described in few animals of genotypes expected to be resistant and, on the other hand , animals expected to be highly susceptible have remained healthy. It is thus important to understand why these cases occur in order to effectively control disease outbreaks and avoid any chance of scrapie appearing in flocks bred for resistance. Previous studies of experimental scrapie have suggested that there may be different strains of scrapie which target different PrP genotypes and the same may be true for natural scrapie. Strain differences can be revealed by transmission to mice and by the biochemical characteristics of PrPsc extracted from affected animals.

 The objectives are firstly to create a network of sheep PrP geneticists throughout Europe and to carry out genotyping at the three most important codons in sheep from different countries and breeds to those which have been tested previously. In order to underpin this effort, improved genotyping techniques will be developed. Strain typing transmission studies (to mice) will be set up from selected outbreaks of scrapie in different countries and comparisons made with previous transmissions using scrapie and BSE. In addition PrPsc biochemical strain typing methods will be improved and differences in brain areas damaged by disease will be assessed. The potential for sheep to act as carriers of infection will be investigated using similar techniques. Sheep deviating from the expected response to scrapie will be investigated for any influence of other PrP gene polymorphic variants and genes other than the PrP gene in disease incidence. 

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PL973315 : Development of a novel diagnostic to assist quality assurance procedures in European meat production (DIAQAM)

Coordinator: H. Schroeder, Institut fuer Physiologische Chemie, Johannes Gutenberg-Universitaet Mainz (DE)
Participants:
 
• * J. Forrest, Cellular and Environmental Physiology, Scottish Crop Research Institute, Dundee (UK)
• * B. Solomon, Department of Molecular Microbiology and Biotechnology, Tel-Aviv University (IL)
• * D. Barber, Veterinary Services, The Scottish Agricultural Services, Edinburgh (UK)
• * S.J. Holmes, ADGEN Diagnostic Systems,Auchincruive (UK)
• * T. Olsson, Reserach and development, Boule Diagnostics AB (SE)
The objectives of the project are:
 
• 1. Development of a blood test for detecting pre-clinical BSE in live cattle.
• 2. Development of a test using tonsils or eyes and associated nervous tissue for detecting preclinical BSE in newly slaughtered cattle.
• 3. Provision of a kit for further testing throughout Europe. Diagnoses of TSEs, including scrapie of sheep and BSE are currently confirmed posthumously either by histological examination to reveal the presence of spongiform areas of the brain or by immunohistochemical detection of the abnormal prion protein PrPsc, a conformational isomer of the normal PrPc. Conformational change renders it insoluble and partially resistant to degradation by proteinase K. Since the two isoforms cannot be distinguished serologically, discrimination depends on treatment of brain tissue with proteinase K, and subsequent recognition of the residual Pr27-30 fragment with a specific antibody.
Recent experiments with sheep, however, have indicated that scrapie may be detected preclinically by identifying PrPsc in tonsils biopsies instead of brain. Tonsillar tissue may also prove to be of diagnostic value in cattle, but obtaining samples from live animals is less convenient than obtaining blood, and the best solution would be to sample tonsils from newly slaughtered animals. instead of tonsils, which might prove to be free of PrPsc in cattle, the eyes and associated nervous tissue may be used for a test on carcasses. It may also be possible to diagnose BSE from the presence of PrPsc or some new marker in blood. The peptide conformation may have an important role in determining prion toxicity. The increased b-sheet structure was found to correlate with enhanced neurotoxic activity. Neurotoxicity of the prion proteins, extracted from biological samples from sick and suspected animals, on the neural cell culture may be an additional diagnostic tool for detectionof PrPsc in BSE. Identification of the regions on the prion molecules where pathological conformational changes occur may facilitate the design and preparation of antibodies or small compounds that might prevent such transition and may delay or prevent the disease. The consortium consists of six partners (1 DE, 3 GB, 1 IL, 1 SE). The inclusion of two SMEs gives the capability of producing prototype kits which may be used to diagnose BSE throughout Europe. 

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PL973308 : Separation, identification and characterization of the normal and abnormal isoforms of prion protein from normal and experimentally infected fish

Coordinator: C.G. Bolis, Institute of Pharmacological Sciences, Faculty of Pharmacy, University of Milan (IT)
Participants:
 
• * S. Ronchi, Istituto di Fisiologia Veterinaria e Biochimica, Faculty of Veterinary Medicine, University of Milan (IT)
• * B. Lahlou, Laboratoire de Physiologie Cellulaire et Molculaire UMR CNRS, Universit de Nice Sophia Antipolis, Nice (FR)
• * A. Figueras, Instituto de Ciencias Marinas, Consejo Superior de Investigaciones Cintificas, Vigo (ES)
• * M. Pocchiari, Laboratory of Virology, Istituto Superiore di Sanita, Rome (IT)
• * P. Pietta, Istituto di Tecnologie Biomediche Avanzate, Consiglio Nazionale delle Ricerche, Segrate (IT)
It was recently described the presence of PrP normal isoform in the brain of salmon. This is phylogenetically very important since it was the first time identified in aquatic vertebrate. This finding will be important to contribute to the better understanding of the functions of PrP normal isoform proteins and the interspecies relationship. The presence of normal isoform was very well described including molecular characterization in several species of terrestrial vertebrates. It was, in addition, described in Drosophila (the fruit fly). The main purpose of the study is to separate, to identify and to perform the molecular characterization of the normal isoform of prion protein from normal and experimentally infected fish. 

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Agricultural research Projects adopted following the FAIR specific call for proposals on TSEs PL973311 : Analysis of molecular factors affecting variability in BSE and scrapie susceptibility Coordinator: J. Williams, Roslin Institute (Edinburgh), Division of Molecular Biology (UK) Participants : * L. Ferretti, Consiglio Nazionale delle Ricerche, Istituto per la Difesa e la Valorizzazione del Germoplasma Animale (IT) * I. Olsaker, Norwegian College of Veterinary Medicine Department of Morphology, Genetics and Aquatic Biology (NO) * P. Kitchin, Rosgen Ltd., Limited Company registered in Scotland (UK) This project will examine the genetic factors involved in the susceptibility of sheep to scrapie and whether there are genetic factors involved in the susceptibility of cattle to BSE. Much work has been carried out in sheep on the relationship between particular forms of the PrP gene and prevalence of natural scrapie, with particular forms in defined breeds being associated with high disease incidence (Hoinville et al Vet Rec 136 312). Studies so far have focused on variations in regions of the gene which code for the protein , and in particular on a specific part of the gene - the octapeptide repeat region. No information is available regarding the way the different forms of the gene affect susceptibility to disease or on regions of the gene which regulate its'expression. In addition although breeding experiments have pointed to PrP as the major factor governing progression of scrapie in sheep, there has been no work to formally exclude the involvement of other genes. In cattle despite the seriousness and extent of the BSE epidemic in the UK, little work has been done to examine the genetics of infection and progression of the disease. The coding region of PrP has been sequenced twice and three polymorphisms have been found (Goldmann et al 1991 J. Gen. Virol. 72 201). The study of the effect of the different forms of PrP on BSE susceptibility has proved inconclusive (Neiburgs et al Anim Genet 25 313) while examination of incidence of disease in calves of BSE affected and unaffected cows, is suggestive of a genetic variation in susceptibility to disease (Hoinville et al 1995 Vet. Rec. 136 312). The work that will be undertaken in this project will add to the knowledge of genetic factors involved in the development of scrapie in sheep and will test whether there are genetic factors affecting the susceptibility of cattle to BSE. The study will focus at several levels: 1. The complete PrP gene including flanking regulatory regions will be sequenced from both sheep and cattle. This information will be used to identify the majority of polymorphisms in the gene for both species. The frequencies of the polymorphisms found will be measured accross different breeds and populations. Selected polymorphisms, focusing on regulatory and conserved elements of the gene, will then be tested for a role in the development of disease in sheep and cattle by linkage analysis. 2. The expression levels of the PrP gene in affected vs unaffected individuals will be examined to see whether the level of PrP expression is associated with pre-disposition to disease. 3. The chromosomal region neighborouring the PrP gene will be examined for other genes that may be involved in disease. This search will initially use of genetic markers, selected from genetic maps, which will be tested for involvement in disease by segregation analysis. In addition novel expressed sequences near to the PrP gene, which may confer differing susceptibility to disease, will be sought through exon trapping using large fragment yeast artificial chromosome (YAC) clones covering the chromosomal region containing the PrP gene. The putative role of the expressed sequences identified in development of scrapie will be tested by examining expression levels in affected vs healthy sheep. 4. Finally the whole genome will be examined for additional loci that are involved in development of BSE in cattle. Using microsatellite markers selected to cover the whole genome at roughly 20 cM intervals (1/150th of the genome) the inheritance of chromosomal regions will be tracted from selected sires into affected and unaffected progeny. Distorted segregation of markers between affected and unaffected progeny will reveal loci that may be involved in the disease. This work will complete the basic molecular information required for studying the disease. Return to top of page PL973314 : Relationship between conformation of PrP, infectivity and pathogenicity of bovine spongiform encephalopathy (BSE) as a basis for diagnosis Coordinator : D. Riesnev, Heinrich-Heine-Universitt Dsseldorf, Institut fr Physikalische Biologie (DE) Participants : * R. Jackman, Veterinary Laboratory Agency, Central Veterinary Laboratory (CVL) Addlestone (UK) * S. Weiss, Ludwig-Maximilians-Universitt Munchen, Institut fur Biochemie -Genzentrum der LMU Mnchen (DE) * D. Dormont, Commissariat  l'Energie Atomique, Direction des Sciences du Vivant Dpartement de Recherche Mdicale, Service de Neurovirologie (FR) * H. Kretzschmar, Georg-August-Universtitt, Institut fr Neuropathologie (DE) * J. Rossier, Centre National de la Recherche Scientifique, Ile de France - Secteur Paris B, Unit de Recherche Associe 2054, Neurobiologie de la Diversit Cellulaire (FR) * C. Reiter, Connex Gesellschaft zur Optimierung von Forschung und Entwicklung Mbh, Connex Gmbh (DE) According to the prion-model, BSE is transmitted by a proteinaceous infectious agent (prion); the major component is a host protein, the so-called prion-protein, which is present in the cellular form, PrPc and after infection in an abnormal form PrPBSE. The abnormal isoform of PrP triggers the transformation of the cellular into the abnormal form thereby multiplying the agent. The knowledge of the basic biological, biochemical, and biophysical features of PrPBSE is regarded as a prerequisite to develop new techniques for diagnosis. The molecular properties of PrPc, PrPBSE and PrP-peptides of the bovine system will be determined and compared with those of scrapie and CJD to differentiate between general principals of prion diseases and particular BSE-features like the so-called BSE-strain of the prion agent. Methods from molecular biology, biochemistry and biophysics will be applied. As an alternative way of a fast and reliable diagnosis of BSE, a cell culture susceptible to BSE infection will be established by transfection of the bovine PrP. Specified antibodies and other PrP-interacting ligands will be studied systematically to differentiate between PrPc and PrPBSE. The mechanism of neurodegeneration and the appearance of early markers after BSE infection will be studied in neuronal cell cultures. With the progress of the project the partners will study the relation between BSE infection, PrP accumulation, PrP conformation, interacting ligands, appearance of early markers and develop an optimal concept for a sensitive diagnosis. Return to top of page PL973304: Generation of bovine and ovine PrP transgenic mice for the development of improved bioassays for BSE and scrapie agent detection Coordinator: M.H. Groschup, Federal Research for Virus Diseases of Animals , Institute for Vaccines, Federal Research Centre for Virus Diseases of Animals (DE) Participants : * M. Dawson, Veterinary Laboratories Agency, Virology Department, Research and Development Division, Addleston (UK) * J.L. Vilotte, Institut National de la Recherche Agronomique - Btiment des Biotechnologies (FR) The need for more sensitive and cost efficient bioassay models for BSE and scrapie infectivity than currently exist is evident. The aim of this project is to generate transgenic mice which highly overexpress bovine and ovine prion protein. All transgenic mice will be analyzed immunochemically with respect to PrPc expression, tissue specificity of expression and histopathologically for spontaneous neuro- and myodegeneration in aged animals. According to the prion theory similarity in the amino acid sequence of the prion proteins of a donor in comparison to the receptor species largely determines the efficiency of transmission of an infectious agent. Accordingly transgenic mice expressing ruminant prion proteins should be more susceptible to BSE or scrapie than conventional mice and therefore useful for the development of highly sensitive bioassays for agent detection. Bovine and ovine prion protein transgenic mice will therefore be test inoculated with brain homogenates of BSE and scrapie diseased animals in order to determine their susceptibility. If animals come down with disease they will be analyzed using histopathology, immunocytochemestry and immunoblot for detection of spongiform brain lesions and accumulation of pathological prion protein. Transgenic mice will eventually be challenged with BSE brain homogenates, of which the inherent infectivity has already been determined by cattle inoculation experiments. Return to top of page PL973306 : New approaches to the diagnosis and control of transmissible spongiform encephalopathies Coordinator: M. Rogers, University College Dublin, University Dept. of Zoology and Biotechnology Centre (IE) Partipicants : * J.M. S!nchez-Vizca"no, Instituto Nacional de Investigaci#n y Tecnolog"a Agraria y Alimentaria, Centro de Investigaci#n en Sanidad Animal (CISA) (ES) * B. Adair, The Queen's University of Belfast, Department of Veterinary Sciences (UK) * T. Linne, Swedish University of Agricultural Sciences, Department of Veterinary Microbiology, Section of Virology - Unit of Molecular Virology (SE) * M. Merza, Svanova Biotech (SE) This project takes a comprehensive approach to the development of diagnostic tests for BSE and other TSEs. Specifically, the proposal seeks: * to develop diagnostic tests for BSE and other TSEs that can be used to detect animals incubating the disease but which do not display clinical symptoms both in a live (field) test and in the tissues of animal carcasses, in an abattoir test. A preclinical test for BSE will either identify PrPsc or some other disease specific marker. Initially, the best candidate marker is PrPsc though this protein is poorly immunogenic and no antibodies are available which can discriminate between the normal and disease specific isoforms. The proposal sets out novel methods that will allow the isolation of PrPsc monoclonal antibodies. Once appropiate antibodies against PrPsc or other marker proteins have been identified, assays using these reagents will be developed and optimised. * to develop transgenic mice expressing PrP constructs from bovine and porcine species that provide rapid methods for assessing the levels of infectious agent in animal tissues or their products. Such animals should provide sensitive and reasonably rapid tests for BSE in different tissues and fluids derived from potentially infected animals. * to determine the relative efficiencies of transmission of BSE to and between pigs by the oral and intracerebral routes as part of the more general investigation into TSEs. Part of this work will also utilise a transgenic mouse model expressing the porcine PrP gene analogous to the bovine model described above. In conclusion, this project presents a comprehensive and complete approach to the development of diagnostic tests for BSE and as part of their development and validation, provides sensitive animal models for the relatively rapid testing of beef and beef products for BSE. Return to top of page PL973301 : Measures to reduce contamination of meat and environment with CNS tissue during slaughter and processing of cattle and sheep Coordinator : D. Harbour, Department of Clinical Veterinary Science, Division of Molecular and Cellular Biology, University of Bristol (UK) Participants : * J.P. Frencia, Association pour le Dvelopment de l'Institut de la Viande, Clermont Ferrand(FR) * J. Sheridan, Teagasc, the National Food Centre, Dublin (IE) * D. Tinker, Silsoe Research Institute, Company Ltd, Bedford (UK) * M. Owen, Meat and Livestock Commission, Public Corporation, Milton Keynes (UK). Innovative techniques to assess, directly or indirectly, the level of cross-contamination with CNS material during conventional slaughter, dressing and butchery of cattle and sheep, will be developed. For direct measurement, nucleic acid-based (RT-PCR) and protein antigen-based (ELISA) assays will be developed to detect CNS-specific mRNA and proteins respectively. Furthermore, an innovative technique to detect prion protein (PrP), based on the affinity of PrP for molecular chaperone proteins, will be developed. For indirect measurement of cross-contamination, the use of tracer dyes and Escherichia coli K12 injected into the CNS will be assessed. These direct and indirect assays will be used to monitor cross-contamination of carcasses, equipment and abattoir environment (including atmospheric) with CNS material during conventional slaughter, dressing and butchery of cattle and sheep, in order to identify critical practices. Based on the results of these studies, changes to conventional methods will be devised, and tested under experimental conditions to ascertain wether cross-contamination can be reduced. Concurrently, innovative equipment (particularly saws and vacuum devices) for the safe removal of CNS material will be developed and tested for their ability to reduce or eliminate cross-contamination. Practices or equipment that do decrease the risks under experimental conditions will be assessed under commercial conditions for economic and practical viability. New practices or equipment will be assessed and demonstrated to industry. A set of guidelines for avoidance of CNS cross-contamination in the meat industry will be drafted for presentation to the EC. Return to top of page PL973305 : Improving prospects for scrapie control in sheep and goats by study of host genotypes, TSE isolates and their in vivo and in vitro interaction Coordinator: J.M Elsen, Institut National de la Recherche Agronomique, Station d'Amlioration Gntique des Animaux, Auzeville (FR) Participants : * M. Dawson, Veterinary Laboratories Agency, Virology Department, Research and Development Division, Addlestone (UK) * N. Hunter, Institute for Animal Health, BBSRC and MRC Neuropathogenesis Unit, Edinburgh (UK) * M.A. Smits, DLO Institute for Animal Science and Health, Lelystad (NL) * M. Groschup, Federal Research Centre for Virus Diseases of Animals, Institute for Vaccines at the Federal Research Centre for Virus Diseases of Animals,Tbingen(DE) * A. Palsdottir, Institute of Experimental Pathology, University of Iceland, Keldur (IS) * O. Papadopoulos, Laboratory of Microbiology and Infectious Diseases Faculty of Veterinary Medicine Aristotle University, Thessaloniki (GR) * J. Roche, University College Dublin, Department of Animal Husbandry and Production (IE) * J. Jarp, Department of Epidemiology, National Veterinary Institute,Oslo (NO) * T. Baron, Laboratoire de Pathologie Bovine, Centre National d'Etudes Vtrinaires et Alimentaires, Lyon (FR) * M.Y. Boscher, Laboratoire d'Analyses Gntiques pour les Esp ces Animales, Jouy En Josas, (FR) * P. Economides, Central veterinary Laboratory, Department of Veterinary Services, Government Services, Nicosia (CY) Differences in sheep PrP amino acid 136, 154 and 171 codons are clearly associated with susceptibility to scrapie in most outbreaks and this has opened the opportunity to breed for resistance to scrapie. However, scrapie has been described in few animals of genotypes expected to be resistant and, on the other hand , animals expected to be highly susceptible have remained healthy. It is thus important to understand why these cases occur in order to effectively control disease outbreaks and avoid any chance of scrapie appearing in flocks bred for resistance. Previous studies of experimental scrapie have suggested that there may be different strains of scrapie which target different PrP genotypes and the same may be true for natural scrapie. Strain differences can be revealed by transmission to mice and by the biochemical characteristics of PrPsc extracted from affected animals. The objectives are firstly to create a network of sheep PrP geneticists throughout Europe and to carry out genotyping at the three most important codons in sheep from different countries and breeds to those which have been tested previously. In order to underpin this effort, improved genotyping techniques will be developed. Strain typing transmission studies (to mice) will be set up from selected outbreaks of scrapie in different countries and comparisons made with previous transmissions using scrapie and BSE. In addition PrPsc biochemical strain typing methods will be improved and differences in brain areas damaged by disease will be assessed. The potential for sheep to act as carriers of infection will be investigated using similar techniques. Sheep deviating from the expected response to scrapie will be investigated for any influence of other PrP gene polymorphic variants and genes other than the PrP gene in disease incidence. Return to top of page PL973315 : Development of a novel diagnostic to assist quality assurance procedures in European meat production (DIAQAM) Coordinator: H. Schroeder, Institut fuer Physiologische Chemie, Johannes Gutenberg-Universitaet Mainz (DE) Participants: * J. Forrest, Cellular and Environmental Physiology, Scottish Crop Research Institute, Dundee (UK) * B. Solomon, Department of Molecular Microbiology and Biotechnology, Tel-Aviv University (IL) * D. Barber, Veterinary Services, The Scottish Agricultural Services, Edinburgh (UK) * S.J. Holmes, ADGEN Diagnostic Systems,Auchincruive (UK) * T. Olsson, Reserach and development, Boule Diagnostics AB (SE) The objectives of the project are: 1. Development of a blood test for detecting pre-clinical BSE in live cattle. 2. Development of a test using tonsils or eyes and associated nervous tissue for detecting preclinical BSE in newly slaughtered cattle. 3. Provision of a kit for further testing throughout Europe. Diagnoses of TSEs, including scrapie of sheep and BSE are currently confirmed posthumously either by histological examination to reveal the presence of spongiform areas of the brain or by immunohistochemical detection of the abnormal prion protein PrPsc, a conformational isomer of the normal PrPc. Conformational change renders it insoluble and partially resistant to degradation by proteinase K. Since the two isoforms cannot be distinguished serologically, discrimination depends on treatment of brain tissue with proteinase K, and subsequent recognition of the residual Pr27-30 fragment with a specific antibody. Recent experiments with sheep, however, have indicated that scrapie may be detected preclinically by identifying PrPsc in tonsils biopsies instead of brain. Tonsillar tissue may also prove to be of diagnostic value in cattle, but obtaining samples from live animals is less convenient than obtaining blood, and the best solution would be to sample tonsils from newly slaughtered animals. instead of tonsils, which might prove to be free of PrPsc in cattle, the eyes and associated nervous tissue may be used for a test on carcasses. It may also be possible to diagnose BSE from the presence of PrPsc or some new marker in blood. The peptide conformation may have an important role in determining prion toxicity. The increased b-sheet structure was found to correlate with enhanced neurotoxic activity. Neurotoxicity of the prion proteins, extracted from biological samples from sick and suspected animals, on the neural cell culture may be an additional diagnostic tool for detectionof PrPsc in BSE. Identification of the regions on the prion molecules where pathological conformational changes occur may facilitate the design and preparation of antibodies or small compounds that might prevent such transition and may delay or prevent the disease. The consortium consists of six partners (1 DE, 3 GB, 1 IL, 1 SE). The inclusion of two SMEs gives the capability of producing prototype kits which may be used to diagnose BSE throughout Europe. Return to top of page PL973308 : Separation, identification and characterization of the normal and abnormal isoforms of prion protein from normal and experimentally infected fish Coordinator: C.G. Bolis, Institute of Pharmacological Sciences, Faculty of Pharmacy, University of Milan (IT) Participants: * S. Ronchi, Istituto di Fisiologia Veterinaria e Biochimica, Faculty of Veterinary Medicine, University of Milan (IT) * B. Lahlou, Laboratoire de Physiologie Cellulaire et Molculaire UMR CNRS, Universit de Nice Sophia Antipolis, Nice (FR) * A. Figueras, Instituto de Ciencias Marinas, Consejo Superior de Investigaciones Cintificas, Vigo (ES) * M. Pocchiari, Laboratory of Virology, Istituto Superiore di Sanita, Rome (IT) * P. Pietta, Istituto di Tecnologie Biomediche Avanzate, Consiglio Nazionale delle Ricerche, Segrate (IT) It was recently described the presence of PrP normal isoform in the brain of salmon. This is phylogenetically very important since it was the first time identified in aquatic vertebrate. This finding will be important to contribute to the better understanding of the functions of PrP normal isoform proteins and the interspecies relationship. The presence of normal isoform was very well described including molecular characterization in several species of terrestrial vertebrates. It was, in addition, described in Drosophila (the fruit fly). The main purpose of the study is to separate, to identify and to perform the molecular characterization of the normal isoform of prion protein from normal and experimentally infected fish. 

---

Agricultural research
Projects = adopted following the FAIR specific call
for proposals on = TSEs
PL973311 : Analysis of molecular factors affecting variability = in BSE and
scrapie
susceptibility
Coordinator: J. Williams, = Roslin Institute (Edinburgh), Division of
Molecular Biology = (UK)
Participants :
* L. Ferretti, Consiglio Nazionale delle = Ricerche, Istituto per la Difesa e
la Valorizzazione del Germoplasma = Animale (IT)
* I. Olsaker, Norwegian College of Veterinary Medicine = Department of
Morphology, Genetics and Aquatic Biology (NO)
* P. = Kitchin, Rosgen Ltd., Limited Company registered in Scotland (UK) =
This project will examine the genetic factors involved in = the
susceptibility of sheep to scrapie and whether there are genetic = factors
involved in the susceptibility of cattle to BSE.
Much work = has been carried out in sheep on the relationship between
particular = forms of the PrP gene and prevalence of natural scrapie, = with
particular forms in defined breeds being associated with high = disease
incidence (Hoinville et al Vet Rec 136 312). Studies so far = have focused on
variations in regions of the gene which code for the = protein , and in
particular on a specific part of the gene - the = octapeptide repeat region.
No information is available regarding the = way the different forms of the
gene affect susceptibility to disease = or on regions of the gene which
regulate its'expression. In addition = although breeding experiments have
pointed to PrP as the major factor = governing progression of scrapie in
sheep, there has been no work to = formally exclude the involvement of other
genes.
In cattle despite = the seriousness and extent of the BSE epidemic in the UK,
little work = has been done to examine the genetics of infection and
progression of = the disease. The coding region of PrP has been sequenced
twice and = three polymorphisms have been found (Goldmann et al 1991 J. = Gen.
Virol. 72 201). The study of the effect of the different forms = of PrP on
BSE susceptibility has proved inconclusive (Neiburgs et al = Anim Genet 25
313) while examination of incidence of disease in = calves of BSE affected
and unaffected cows, is suggestive of a = genetic variation in susceptibility
to disease (Hoinville et al 1995 = Vet. Rec. 136 312).
The work that will be undertaken in this project = will add to the knowledge
of genetic factors involved in the = development of scrapie in sheep and will
test whether there are = genetic factors affecting the susceptibility of
cattle to BSE. The = study will focus at several levels:
1. The complete PrP gene = including flanking regulatory regions will be
sequenced from both = sheep and cattle. This information will be used to
identify the = majority of polymorphisms in the gene for both species. = The
frequencies of the polymorphisms found will be measured accross = different
breeds and populations. Selected polymorphisms, focusing on = regulatory and
conserved elements of the gene, will then be tested = for a role in the
development of disease in sheep and cattle by = linkage analysis.

2. The expression levels of the PrP gene in = affected vs unaffected
individuals will be examined to see whether = the level of PrP expression is
associated with pre-disposition to = disease.

3. The chromosomal region neighborouring the PrP gene = will be examined for
other genes that may be involved in disease. = This search will initially use
of genetic markers, selected from = genetic maps, which will be tested for
involvement in disease by = segregation analysis. In addition novel expressed
sequences near to = the PrP gene, which may confer differing susceptibility
to disease, = will be sought through exon trapping using large fragment = yeast
artificial chromosome (YAC) clones covering the chromosomal = region
containing the PrP gene. The putative role of the expressed = sequences
identified in development of scrapie will be tested by = examining expression
levels in affected vs healthy sheep.

4. = Finally the whole genome will be examined for additional loci that = are
involved in development of BSE in cattle. Using microsatellite = markers
selected to cover the whole genome at roughly 20 cM intervals = (1/150th of
the genome) the inheritance of chromosomal regions will = be tracted from
selected sires into affected and unaffected progeny. = Distorted segregation
of markers between affected and unaffected = progeny will reveal loci that
may be involved in the disease. =
This work will complete the basic molecular information required = for
studying the disease.
Return to top of page
PL973314 : = Relationship between conformation of PrP, infectivity = and
pathogenicity of bovine spongiform encephalopathy (BSE) as a = basis for
diagnosis
Coordinator : D. Riesnev, = Heinrich-Heine-Universit=E4t D=FCsseldorf, Institut
f=FCr = Physikalische Biologie (DE)
Participants :
* R. Jackman, = Veterinary Laboratory Agency, Central Veterinary Laboratory
(CVL) = Addlestone (UK)
* S. Weiss, Ludwig-Maximilians-Universit=E4t = Munchen, Institut fur Biochemie
-Genzentrum der LMU M=FCnchen (DE) =
* D. Dormont, Commissariat =E0 l'Energie Atomique, Direction des = Sciences du
Vivant D=E9partement de Recherche M=E9dicale, Service de = Neurovirologie (FR)
* H. Kretzschmar, Georg-August-Universtit=E4t, = Institut f=FCr Neuropathologie
(DE)
* J. Rossier, Centre National = de la Recherche Scientifique, Ile de France -
Secteur Paris B, = Unit=E9 de Recherche Associ=E9e 2054, Neurobiologie de la
Diversit=E9 = Cellulaire (FR)
* C. Reiter, Connex Gesellschaft zur Optimierung von = Forschung und
Entwicklung Mbh, Connex Gmbh (DE)
According to the = prion-model, BSE is transmitted by a proteinaceous
infectious agent = (prion); the major component is a host protein, the
so-called = prion-protein, which is present in the cellular form, PrPc and
after = infection in an abnormal form PrPBSE. The abnormal isoform of = PrP
triggers the transformation of the cellular into the abnormal = form thereby
multiplying the agent. The knowledge of the basic = biological, biochemical,
and biophysical features of PrPBSE is = regarded as a prerequisite to develop
new techniques for = diagnosis.
The molecular properties of PrPc, PrPBSE and PrP-peptides = of the bovine
system will be determined and compared with those of = scrapie and CJD to
differentiate between general principals of prion = diseases and particular
BSE-features like the so-called BSE-strain of = the prion agent. Methods from
molecular biology, biochemistry and = biophysics will be applied. As an
alternative way of a fast and = reliable diagnosis of BSE, a cell culture
susceptible to BSE = infection will be established by transfection of the
bovine PrP. = Specified antibodies and other PrP-interacting ligands will = be
studied systematically to differentiate between PrPc and PrPBSE. = The
mechanism of neurodegeneration and the appearance of early = markers after
BSE infection will be studied in neuronal cell = cultures. With the progress
of the project the partners will study = the relation between BSE infection,
PrP accumulation, PrP = conformation, interacting ligands, appearance of
early markers and = develop an optimal concept for a sensitive diagnosis.
Return to top = of page
PL973304: Generation of bovine and ovine PrP transgenic mice = for the
development of improved bioassays for BSE and scrapie agent = detection
Coordinator: M.H. Groschup, Federal Research for Virus = Diseases of Animals
, Institute for Vaccines, Federal Research Centre = for Virus Diseases of
Animals (DE)
Participants :
* M. Dawson, = Veterinary Laboratories Agency, Virology Department, Research
and = Development Division, Addleston (UK)
* J.L. Vilotte, Institut = National de la Recherche Agronomique - B=E2timent
des Biotechnologies = (FR)
The need for more sensitive and cost efficient bioassay models = for BSE and
scrapie infectivity than currently exist is evident. The = aim of this
project is to generate transgenic mice which highly = overexpress bovine and
ovine prion protein. All transgenic mice will = be analyzed immunochemically
with respect to PrPc expression, tissue = specificity of expression and
histopathologically for spontaneous = neuro- and myodegeneration in aged
animals. According to the prion = theory similarity in the amino acid
sequence of the prion proteins of = a donor in comparison to the receptor
species largely determines the = efficiency of transmission of an infectious
agent. Accordingly = transgenic mice expressing ruminant prion proteins
should be more = susceptible to BSE or scrapie than conventional mice and
therefore = useful for the development of highly sensitive bioassays for
agent = detection. Bovine and ovine prion protein transgenic mice = will
therefore be test inoculated with brain homogenates of BSE and = scrapie
diseased animals in order to determine their susceptibility. = If animals
come down with disease they will be analyzed using = histopathology,
immunocytochemestry and immunoblot for detection of = spongiform brain
lesions and accumulation of pathological prion = protein. Transgenic mice
will eventually be challenged with BSE brain = homogenates, of which the
inherent infectivity has already been = determined by cattle inoculation
experiments.
Return to top of = page
PL973306 : New approaches to the diagnosis and control of = transmissible
spongiform encephalopathies
Coordinator: M. Rogers, = University College Dublin, University Dept. of
Zoology and = Biotechnology Centre (IE)
Partipicants :
* J.M. = S=E1nchez-Vizca=EDno, Instituto Nacional de Investigaci=F3n y = Tecnolog=EDa
Agraria y Alimentaria, Centro de Investigaci=F3n en = Sanidad Animal (CISA)
(ES)
* B. Adair, The Queen's University of = Belfast, Department of Veterinary
Sciences (UK)
* T. Linne, = Swedish University of Agricultural Sciences, Department of
Veterinary = Microbiology, Section of Virology - Unit of Molecular Virology
(SE) =
* M. Merza, Svanova Biotech (SE)
This project takes a = comprehensive approach to the development of
diagnostic tests for BSE = and other TSEs. Specifically, the proposal seeks:
* to develop = diagnostic tests for BSE and other TSEs that can be used to
detect = animals incubating the disease but which do not display = clinical
symptoms both in a live (field) test and in the tissues of = animal
carcasses, in an abattoir test. A preclinical test for BSE = will either
identify PrPsc or some other disease specific marker. = Initially, the best
candidate marker is PrPsc though this protein is = poorly immunogenic and no
antibodies are available which can = discriminate between the normal and
disease specific isoforms. The = proposal sets out novel methods that will
allow the isolation of = PrPsc monoclonal antibodies. Once appropiate
antibodies against PrPsc = or other marker proteins have been identified,
assays using these = reagents will be developed and optimised.
* to develop transgenic = mice expressing PrP constructs from bovine and
porcine species that = provide rapid methods for assessing the levels of
infectious agent in = animal tissues or their products. Such animals should
provide = sensitive and reasonably rapid tests for BSE in different tissues
and = fluids derived from potentially infected animals.
* to determine the = relative efficiencies of transmission of BSE to and
between pigs by = the oral and intracerebral routes as part of the more
general = investigation into TSEs. Part of this work will also utilise = a
transgenic mouse model expressing the porcine PrP gene analogous to = the
bovine model described above.
In conclusion, this project = presents a comprehensive and complete approach
to the development of = diagnostic tests for BSE and as part of their
development and = validation, provides sensitive animal models for the
relatively rapid = testing of beef and beef products for BSE.
Return to top of page =
PL973301 : Measures to reduce contamination of meat and environment = with
CNS tissue during slaughter and processing of cattle and = sheep
Coordinator : D. Harbour, Department of Clinical Veterinary = Science,
Division of Molecular and Cellular Biology, University of = Bristol (UK)
Participants :
* J.P. Frencia, Association pour le = D=E9velopment de l'Institut de la Viande,
Clermont Ferrand(FR)
* = J. Sheridan, Teagasc, the National Food Centre, Dublin (IE)
* D. = Tinker, Silsoe Research Institute, Company Ltd, Bedford (UK)
* M. = Owen, Meat and Livestock Commission, Public Corporation, Milton = Keynes
(UK).
Innovative techniques to assess, directly or = indirectly, the level of
cross-contamination with CNS material during = conventional slaughter,
dressing and butchery of cattle and sheep, = will be developed. For direct
measurement, nucleic acid-based = (RT-PCR) and protein antigen-based (ELISA)
assays will be developed = to detect CNS-specific mRNA and proteins
respectively. Furthermore, = an innovative technique to detect prion protein
(PrP), based on the = affinity of PrP for molecular chaperone proteins, will
be developed. = For indirect measurement of cross-contamination, the use of
tracer = dyes and Escherichia coli K12 injected into the CNS will = be
assessed.
These direct and indirect assays will be used to = monitor
cross-contamination of carcasses, equipment and abattoir = environment
(including atmospheric) with CNS material during = conventional slaughter,
dressing and butchery of cattle and sheep, in = order to identify critical
practices. Based on the results of these = studies, changes to conventional
methods will be devised, and tested = under experimental conditions to
ascertain wether cross-contamination = can be reduced. Concurrently,
innovative equipment (particularly saws = and vacuum devices) for the safe
removal of CNS material will be = developed and tested for their ability to
reduce or eliminate = cross-contamination. Practices or equipment that do
decrease the = risks under experimental conditions will be assessed under
commercial = conditions for economic and practical viability.
New practices or = equipment will be assessed and demonstrated to industry. A
set of = guidelines for avoidance of CNS cross-contamination in the = meat
industry will be drafted for presentation to the EC.
Return = to top of page
PL973305 : Improving prospects for scrapie control in = sheep and goats by
study of host genotypes, TSE isolates and their in = vivo and in vitro
interaction
Coordinator: J.M Elsen, Institut = National de la Recherche Agronomique,
Station d'Am=E9lioration = G=E9n=E9tique des Animaux, Auzeville (FR)
Participants :
* M. = Dawson, Veterinary Laboratories Agency, Virology Department, = Research
and Development Division, Addlestone (UK)
* N. Hunter, = Institute for Animal Health, BBSRC and MRC Neuropathogenesis
Unit, = Edinburgh (UK)
* M.A. Smits, DLO Institute for Animal Science and = Health, Lelystad (NL)
* M. Groschup, Federal Research Centre for = Virus Diseases of Animals,
Institute for Vaccines at the Federal = Research Centre for Virus Diseases of
Animals,T=FCbingen(DE)
* A. = Palsdottir, Institute of Experimental Pathology, University = of
Iceland, Keldur (IS)
* O. Papadopoulos, Laboratory of = Microbiology and Infectious Diseases
Faculty of Veterinary Medicine = Aristotle University, Thessaloniki (GR)
* J. Roche, University = College Dublin, Department of Animal Husbandry and
Production (IE) =
* J. Jarp, Department of Epidemiology, National Veterinary = Institute,Oslo
(NO)
* T. Baron, Laboratoire de Pathologie Bovine, = Centre National d'Etudes
V=E9t=E9rinaires et Alimentaires, Lyon (FR) =
* M.Y. Boscher, Laboratoire d'Analyses G=E9n=E9tiques pour les = Esp=E8ces
Animales, Jouy En Josas, (FR)
* P. Economides, Central = veterinary Laboratory, Department of Veterinary
Services, Government = Services, Nicosia (CY)
Differences in sheep PrP amino acid 136, 154 = and 171 codons are clearly
associated with susceptibility to scrapie = in most outbreaks and this has
opened the opportunity to breed for = resistance to scrapie. However, scrapie
has been described in few = animals of genotypes expected to be resistant
and, on the other hand = , animals expected to be highly susceptible have
remained healthy. It = is thus important to understand why these cases occur
in order to = effectively control disease outbreaks and avoid any chance of
scrapie = appearing in flocks bred for resistance. Previous studies = of
experimental scrapie have suggested that there may be different = strains of
scrapie which target different PrP genotypes and the same = may be true for
natural scrapie. Strain differences can be revealed = by transmission to mice
and by the biochemical characteristics of = PrPsc extracted from affected
animals.
The objectives are firstly = to create a network of sheep PrP geneticists
throughout Europe and to = carry out genotyping at the three most important
codons in sheep from = different countries and breeds to those which have
been tested = previously. In order to underpin this effort, improved
genotyping = techniques will be developed. Strain typing transmission studies
(to = mice) will be set up from selected outbreaks of scrapie in = different
countries and comparisons made with previous transmissions = using scrapie
and BSE. In addition PrPsc biochemical strain typing = methods will be
improved and differences in brain areas damaged by = disease will be
assessed. The potential for sheep to act as carriers = of infection will be
investigated using similar techniques. Sheep = deviating from the expected
response to scrapie will be investigated = for any influence of other PrP
gene polymorphic variants and genes = other than the PrP gene in disease
incidence.
Return to top of = page
PL973315 : Development of a novel diagnostic to assist quality = assurance
procedures in European meat production = (DIAQAM)
Coordinator: H. Schroeder, Institut fuer Physiologische = Chemie, Johannes
Gutenberg-Universitaet Mainz = (DE)
Participants:
* J. Forrest, Cellular and Environmental = Physiology, Scottish Crop Research
Institute, Dundee (UK)
* B. = Solomon, Department of Molecular Microbiology and = Biotechnology,
Tel-Aviv University (IL)
* D. Barber, Veterinary = Services, The Scottish Agricultural Services,
Edinburgh (UK)
* = S.J. Holmes, ADGEN Diagnostic Systems,Auchincruive (UK)
* T. Olsson, = Reserach and development, Boule Diagnostics AB (SE)
The objectives = of the project are:
1. Development of a blood test for detecting = pre-clinical BSE in live
cattle.

2. Development of a test = using tonsils or eyes and associated nervous
tissue for detecting = preclinical BSE in newly slaughtered cattle.

3. Provision of a = kit for further testing throughout Europe.
Diagnoses of TSEs, = including scrapie of sheep and BSE are currently
confirmed = posthumously either by histological examination to reveal = the
presence of spongiform areas of the brain or by = immunohistochemical
detection of the abnormal prion protein PrPsc, a = conformational isomer of
the normal PrPc. Conformational change = renders it insoluble and partially
resistant to degradation by = proteinase K. Since the two isoforms cannot be
distinguished = serologically, discrimination depends on treatment of brain
tissue = with proteinase K, and subsequent recognition of the residual
Pr27-30 = fragment with a specific antibody.
Recent experiments with sheep, = however, have indicated that scrapie may be
detected preclinically by = identifying PrPsc in tonsils biopsies instead of
brain. Tonsillar = tissue may also prove to be of diagnostic value in cattle,
but = obtaining samples from live animals is less convenient than = obtaining
blood, and the best solution would be to sample tonsils = from newly
slaughtered animals. instead of tonsils, which might prove = to be free of
PrPsc in cattle, the eyes and associated nervous tissue = may be used for a
test on carcasses. It may also be possible to = diagnose BSE from the
presence of PrPsc or some new marker in = blood.
The peptide conformation may have an important role in = determining prion
toxicity. The increased =DF-sheet structure was = found to correlate with
enhanced neurotoxic activity. Neurotoxicity = of the prion proteins,
extracted from biological samples from sick = and suspected animals, on the
neural cell culture may be an = additional diagnostic tool for detectionof
PrPsc in BSE. = Identification of the regions on the prion molecules = where
pathological conformational changes occur may facilitate the = design and
preparation of antibodies or small compounds that might = prevent such
transition and may delay or prevent the disease.
The = consortium consists of six partners (1 DE, 3 GB, 1 IL, 1 SE). = The
inclusion of two SMEs gives the capability of producing prototype = kits
which may be used to diagnose BSE throughout Europe.
Return = to top of page
PL973308 : Separation, identification and = characterization of the normal
and abnormal isoforms of prion protein = from normal and experimentally
infected fish
Coordinator: C.G. = Bolis, Institute of Pharmacological Sciences, Faculty of
Pharmacy, = University of Milan (IT)
Participants:
* S. Ronchi, Istituto di = Fisiologia Veterinaria e Biochimica, Faculty of
Veterinary Medicine, = University of Milan (IT)
* B. Lahlou, Laboratoire de Physiologie = Cellulaire et Mol=E9culaire UMR CNRS,
Universit=E9 de Nice Sophia = Antipolis, Nice (FR)
* A. Figueras, Instituto de Ciencias Marinas, = Consejo Superior de
Investigaciones Ci=E9ntificas, Vigo (ES)
* M. = Pocchiari, Laboratory of Virology, Istituto Superiore di Sanita, = Rome
(IT)
* P. Pietta, Istituto di Tecnologie Biomediche = Avanzate, Consiglio
Nazionale delle Ricerche, Segrate (IT)
It was recently described the presence of PrP normal isoform in = the brain
of salmon. This is phylogenetically very important since it = was the first
time identified in aquatic vertebrate.
This finding = will be important to contribute to the better understanding of
the = functions of PrP normal isoform proteins and the = interspecies
relationship.
The presence of normal isoform was very = well described including molecular
characterization in several = species of terrestrial vertebrates. It was, in
addition, described in = Drosophila (the fruit fly).
The main purpose of the study is to = separate, to identify and to perform
the molecular characterization = of the normal isoform of prion protein from
normal and experimentally = infected fish.
 
 
 
 

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Multidisciplinary Scientific Committee / = Scientific Steering Committee (MDSC/SSC)

Outcome of discussions

Summary Minutes - Meeting of 16 October = 1997

1. Welcome, apologies, introductory remarks

The chairman Professor F.Kemper welcomed the participants and = opened the meeting at 10h00.

The list of participants is attached as annex 1. Apologies were = received from: Prof. D.Dormont, Prof.M.Pocchiari, Prof.G.Vicari, and = Prof.M.Wierup.

2. Approval of the agenda and modifications

The agenda was approved with the following modifications:

• Point I.3. (Tallow and tallow derivatives: examination of newly = received information and requests for clarification on the opinion = adopted on 8.09.97.). It was agreed not to discuss this point as such, = but to refer it to a special working group. The discussion on the = mandate and composition of the working group would be discussed under = point I.3.
• Point III.3. (Report by the services of the Commission). The = following item was included: "report by the Joint Research Centre on the = validation study of methods for identifying animal proteins in MBM and = for verifying the temperatures which were applied when producing the = MBM."
The adopted agenda is attached as annex 2.

3. Approval of the minutes of the meeting of 8 September 1997

The minutes of the meeting of 8 September 1997 were adopted with = minor modifications.

Part I: Matters related to BSE

I.1. Specified Risk Materials: further discussion and possibly = adoption of the proposed opinion which was redrafted on the basis of the = discussions of 8 September 1997 (This point includes the discussion = on the question whether the intestine should be considered to be a high = risk tissue)

Following the MDSC/SSC meeting of 8.09.97, all MDSC/SSC members had = been consulted in writing (This written consultation was held between = 24.09. and 15.10.97) on a draft opinion re-written on the basis of the = elements of the discussions of 8.09.97. The comments resulting from this = consultation were at their turn used to prepare an updated version of = that draft opinion. This latest draft served as the basis of the = discussions of 16.10.97.

From the in-depth discussions it became clear that issues such as the = definition ad listing/categorising of Specified Risk Materials, and more = generally the issue of product safety (tallow, meat and bone meal, = gelatine, etc.), probably needed to be addressed differently depending = upon:

• the material itself (part of the animal; tissue, organ or fluid; = .....);
• the animal species (in practice: bovines; ovines and / or caprines); =
• the origin of the material (sourcing and TSE status of a country or = herd, .....);
• the age of the animal,
• the production process to which the materials will be = submitted;
and possibly exploited differently depending upon:
• the use that will be made of the final product: feed and food; = technical or industrial; pharmaceutical (oral intake); pharmaceutical = (other ways of application); intermediate product used as raw material = for another production process; etc.
The preceding principle asks for a methodological approach which is = different from the approach followed so far by the MDSC and other = Scientific Committees when addressing BSE related issues.

The issue was therefore referred for further detailed study to a = working group, to be chaired and reported on by Prof.James. The working = group is expected to submit, before 10 November 1997, a draft report to = Scientific Steering Committee which will discuss it during its plenary = session of 21 November 1997.

The following names were proposed as possible members of the working = group: Prof.P.James (chairperson and rapporteur), Prof.P.Willeberg, = Prof. R.G.Will, Dr. R.Bradley, Prof.A.Somogyi, Prof.D.Dormont, = Dr.Dieringer, Prof.G.Vicari. Prof.P.James accepted the chairmanship.

I.2. Meat and Bone Meal: discussion and possibly adoption of an = opinion on the minimum production process conditions required for = inactivating / eliminating the BSE infective agent.

From the above written consultation (see previous item I.1.), is = became also clear that issues such as the safety of tallow, MBM, = gelatine, etc. may have to be addressed differently depending upon the = use that will be made of the final product, the source of the raw = material, the quality of the raw material, the animal species, the age = of the animal, the production process, etc.

Also, the final report of the 1991-1997 Rendering Study, referred to = in many discussions and frequently used as the scientific basis for the = '133=B0C/20'/3 bar' norm, had become available just a few days before = the meeting and was first distributed in the beginning of the meeting. = Although the essential results remain the same, the report provides more = detail on the alternative methods tested, etc.;

It was therefore decided, and in spite of the fact that a draft = opinion was available which integrated the comments received during the = written consultation procedure, to refer also this issue to a = specialised working group.

Also this working group is expected to submit before 21 November 1997 = a detailed draft report, including a completed matrix representing the = "expected degree of remaining infectivity and/or infectious risk of the = final product (Please note that a raw input material in a given = production process can be a final product from another process) as a = function of use / source (May include: geographical source; herd of = origin; age of the animal; animal species; ....) of the raw material / = quality of the raw material (can be expressed in various ways, for = example "Specified Risk Material belonging to infectivity class X"; or = "material with a high, medium, low and zero level of infectivity". etc.) = / production process". The Scientific Steering Committee will discuss = the report during its plenary session of 21 November 1997.

The following names were proposed as possible members of the working = group: Prof.M.Vanbelle (chairperson and rapporteur), Prof.G.Piva, = Prof.D.Dormont, Prof.M.Taylor, Prof. M.Wierup. Prof.Vanbelle accepted = the chairmanship.

Note: The above working group will also address the tallow = issue (see item I.3 hereafter).

I.3. Tallow and tallow derivatives: examination of newly = received information and requests for clarification on the opinion = adopted on 8.09.97.

The chairman informed the meeting on the following new elements, = which occurred since the MDSC/SSC meeting of 8 September 1997:

a) A number of industry associations and third countries submitted = dossiers defending the thesis that the normal industrial tallow = production processes - even the ones using the lowest time/temperature = combinations - and corresponding research have shown to result in a = product which is free from detectable TSE infectivity, even if the = source material was highly infective. The explanation given is that = because of the proteinaceous nature of the TSE agents they would tend to = remain with the cellular residues of meat and bone meal during = extraction process, rather than be extracted with the lipids of tallow. = (The corresponding documents are: MDSC/DGXXIV/116, 117, 118, 119, = 120,136.)

b) Meanwhile, also the detailed final report on the 1991-1997 = rendering study became available on 6 October and was distributed to all = members. This part of the results of this study which concern tallow are = often used as a justification why tallow can be considered safe even if = it had been submitted during the production process to much less harsh = conditions.

c) From the above written consultation (see item I.1), is became also = clear that issues such as the safety of tallow, MBM, gelatine, etc. may = have to be addressed differently depending upon the use that will be = made of the final product, the source of the raw material, etc.

The chairman therefore proposed that item I.3. "Examination of = newly received information and requests for clarification on the opinion = adopted on 8.09.97" be handled in detail by a subgroup of = specialists. This proposal was accepted and, keeping in mind the = discussions on specified risk materials (see item I.1.) and meat and = bone meal (see item I.2.), it was proposed that its exact mandate would = be phrased in a general way similar to the meat and bone meal one and = taken up by the same group of specialists addressing also the meat and = bone meal issue. The working group is expected to submit a draft report = to Scientific Steering Committee which will discuss it during its = plenary session of 21 November 1997.

I.4.Maternal transmission of BSE: additional questions on the = follow up of the opinion of 8.09.97; planning and approach to prepare an = opinion on these questions.

In the light of the scientific opinions of the Scientific = Veterinary Committee (ScVC) of 17 February 1997 and of the MDSC/SSC of = 16.05.97 that '(....) Bovine milk from healthy cows and products = derived there from which contain no animal derived additives can be = safely consumed in any form by any species. There is no evidence that = milk transmits BSE and the Committee concludes any risk from milk to = be negligible'(....), In the light of the fact that previous opinions = were, amongst several other arguments, based on a conclusion that = (paragraph 3, page 9 of report VI/8197/96 Version J (Final) (This report = corresponds with Document MDSC/DGXXIV/97/014) "There is firm no = evidence to support the occurrence of maternal transmission or the = involvement of milk in the transmission of TME or TSE in other species = of Bovidae or Felidae".

In the light of the scientific opinion of 8.09.97 that the = MDSC/SSC ' (...) supported the general conclusion (...) that a low = level of maternal risk enhancement is likely to exist. The overall = magnitude is estimated at 5 to 15% (95% confidence limits), but with an = increased risk in the months before and after the date of clinical onset = in the dam. This effect is likely due to some form of direct maternal = transmission, although differences in genetic susceptibility cannot be = excluded. (...)',

the opinion of the Multidisciplinary Scientific Committee / = Scientific Steering Committee was requested on:

a) whether or not the discussion on possible risks of consuming milk, = colostrum and milk products, should be re-opened.

b) other possible routes of infection to explain the maternal = transmission of BSE;

c) a risk assessment for these routes;

d) recommendations for options to mitigate the risk from these = routes;

e) recommendations regarding possible activities to be undertaken by = (a) Scientific Committee(s) or other bodies, regarding possible = additional work, further research to be undertaken, encouraged or = funded; regarding further data analyses, etc.

Regarding question a), the MDSC/SSC agreed that the adoption of the = opinion on maternal transmission on 8.09.97, did not have as consequence = that discussion on the safety of milk should be re-opened. The ScVC's = opinion was indeed based on several other major arguments apart from the = one regarding maternal transmission.

Regarding the questions b) through d), Prof.Willeberg highlighted = that the evidence for maternal transmission of BSE used for the opinion = of 8.09.97 comes from two independent data-sets (1) the BSE maternal = cohort study and (2) the BSE case data of the Weybridge epidemiology = group. Both data-sets were statistically analysed but the importance of = the analysis was not in the first place to decide whether or not there = is maternal transmission but more to estimate the extend of maternal = risk enhancement. In terms of population dynamics of BSE in cattle the = effect of maternal risk enhancement is small (the epidemic is expected = to fade out anyway) even when this maternal risk enhancement would = entirely maternal transmission. The possible implications for human = health without an understanding of the mechanisms underlying this = maternal risk enhancement cannot easily be understood.

What precedes implies that questions b), c), d) and e) can not easily = be answered as the mechanisms underlying this maternal risk enhancement = are not (yet fully) known. Answering them would require additional = study, possibly additional research. These questions should therefore be = further discussed by the ad hoc group on BSE of the Scientific Steering = Committee, to be created as soon as possible after the installation of = the latter one, probably towards end November or in the beginning of = December 1997.

I.5. Review of the situation concerning di-calcium phosphate and = amino acids and peptides produced from animal origin;

a) Di-calcium phosphate. Prof.D.Dormont being apologised, = the progress report was delivered by the chairman. In addition to = previous efforts, the MDSC/SSC secretariat sought contact with 12 = companies in Germany, UK, France, Italy, the Netherlands and Belgium. 3 = constructive responses were received, including the documentation = MDSC/DGXXIV/97/104 provided by the association GME - Gelatine = Manufactures of Europe. GME had stated that this documentation was = in fact representative for all production processes in Europe: all = producers of bi-calcium phosphate except one seem indeed to be member of = GME. (The exception is Agfa-Gevaert, that produces its own gelatine for = photographic products). If this is correct, then the problem raised = during the preceding meeting (namely: the available documentation was = not representative for all production processes in all producing Member = States) could be considered as solved. The MDSC/SSC therefore requested = its secretariat to ask GME for written confirmation and evidence that = its members respected the process description provided in the GME = documentation. Such confirmation and evidence could consist of written = declarations from the members and, if appropriate, include a description = how and to what extend local production processes possibly deviate from = the "representative" one described in the GME document).

b) Amino acids and peptides produced from animal origin and = derived products. Prof.G.Pascal, rapporteur, reported on the = progress made. With the help of the MDSC/SSC secretariat he finished the = compilation of a documentation set which would permit him to draft an = opinion on the safety of amino acids and peptides, for discussion and = possible adoption by the SSC. The draft would be made available before = next meeting.

I.6. Information on the Export Certified Herd Scheme and on the = BSE status of Australia, New-Zealand, Norway and the USA: reporting by = the Scientific Veterinary Committee.

Prof. Del Real, chairman of the Scientific Veterinary Committee, = reported on the activities of the BSE subgroup of the ScVC which = analysed the above dossiers and on the plenary session of 16-17.09.97 of = the ScVC during which the subgroup reports were discussed, amended and = approved. Further details can be found in:

• Document N=B0 MDSC/DGXXIV/97/126. CEC, 1997. Report of the = meeting of 16-17.09.97 of the Scientific Veterinary Committee on the = Revised UK Certified Herd Scheme.
• Document N=B0 MDSC/DGXXIV/97/127. CEC, 1997. Report of the = meeting of 16-17.09.97 of the Scientific Veterinary Committee on the TSE = Status of Australia and USA and the Scrapie Eradication Programme in = Norway.
I.7. Possible link between Bovine Spongiform Encephalopathy and = the new variant Creutzfeldt-Jakob Disease (nvCJD): brief discussion.

The chairman briefly introduced the two articles published in = Nature of 2 October 1997. They had been made available to the MDSC/SSC = members as documents N=B0 Document N=B0 MDSC/DGXXIV/97/143, Document = N=B0 MDSC/DGXXIV/97/144 and Document N=B0 MDSC/DGXXIV/97/145.

PART II: Any other business

II.1. Date of the next meeting.

The next meeting has been scheduled for 21 November 1997.

II.2. Miscellaneous:

The chairman reported on an offer received from the industry, to = present to the MDSC/SSC the protocol of a planned new validation study = on the removal/inactivation capacity of the gelatine manufacturing = process with respect to the BSE agent (see document MDSC/DGXXIV/97/106). = The offer was not discussed in detail, given the fact that this was the = last meeting of the MDSC/SSC in its present configuration and with its = present mandate. However, the principle of accepting such offers will = have to be discussed by the Scientific Steering Committee when = establishing its working procedures.

PART III: Matters related to co-ordination

III.1. Establishment of the 8 new Scientific Committees: the = state of affairs;

Mr.Carsin reported on the state of affairs in the selection = procedure for the members of the 8 new Scientific Committees. More then = 800 submissions were received. This corresponded to more than 1000 = expressions of interest, for one could express an interest for more then = one Committee. On 16.10.97, the technical and scientific evaluation of = the candidates had finished and a written procedure was about to be = started to prepare the Commission Decision nominating the members. It = was expected that this procedure would be finalised by the end of = October 1997 and that the Committees may have their first meeting in the = first half of November.

III.2. Reports of the chairs of the at present existing scientific = committees;

Dr. Del Real reported on the activities of the Scientific = Veterinary Committee. During the plenary session of 16-17 September, = 11 reports were adopted, covering the following fields:

• Rabies and movement of animals in the European Union;
• The use of marker vaccines in the control of infectious diseases in = particular, Classical Swine Fever;
• General criteria for Member States eradicating the fish diseases = Viral Haemorrhagic Septicaemia and Infectious Hematopoietic Necrosis; =
• The risk of transferring live fish, their eggs and gametes from = approved farms and minizones in non-approved zones into approved zones; =
• Mechanically recovered meat, health rules applicable to the = production and use of mechanically recovered meat;
• Verocytotoxin-producing Escherichia Coli (VTEC);
• The "Revised UK Certified Herds Scheme";
• The TSE status of Australia and the USA and the Scrapie eradication = programme in Norway;
• The Welfare of Intensively Kept Pigs
• The Killing of Animals for Disease Control Purposes
For information on the Export Certified Herd Scheme and on the BSE = status of Australia, New-Zealand, Norway and the USA, he referred to = item I.7. on the agenda.

Prof. Jouany (Chair, Scientific Committee on Pesticides) = reported on the opinion provided by the Committee on the safety of maize = genetically modified in such way it has become both herbicide resistant = and insect resistant. He stated that given the wide-spread impact of = herbicide resistant plants obtained by genetic modification, the = Scientific Committee for Pesticides considered that each application for = marketing such a plant needs to be evaluated on a case by case = basis.

Prof.Wennig reported on the Scientific Committee = Toxicity/Ecotoxicity of Chemical Compounds and on the opinion on the = possible risk related to the Nickel content of the Euro coins. Results = of tests for nickel release from samples of coins of 2 Euro are now = being circulated. These results indicate that the nickel release from = the Euro coins is in the same range as that from current coins with the = same composition. Pending written comments from the members of the = Committee, Mr Wennig considered that the opinion could thus be = maintained unchanged.

Prof.Vanbelle, chairman of the Scientific Committee Animal = Nutrition briefly reported on the following 9 reports that were = adopted during the 110th plenary meeting of the SCAN on 25-26 September = 1997:

• the use of Corynebacterium melassecola and Brevibacterium = lactofermento as feeding stuffs
• halofuginone for chickens reared for laying
• maduramycin for Turkeys
• formaldehyde
• salinomycin for chickens reared for laying
• diclazuril for turkeys for fattening
• diclazuril for chickens reared for laying
• avilamycin for turkeys for fattening
• the clause of safeguard of Germany against the use of = ronidazole
Prof.Pascal reported on the activities of the Scientific Committee = on Food since the last meeting of the MDSC/SSC (8.09.97). Some of = the major areas covered are:
• the consequences of the maximum pesticide residue levels for the = health of infants, babies and weaning babies.
• the labelling of products containing genetically modified organisms = or derived from it. (Prof. Pascal signalled that this issue is not = exclusively scientific but has some major administrative and legislative = aspects.)
• the maximum acceptable residue levels of Aflatoxin-B1 in food in = relation to the risk for liver cancer.
Prof.Kemper, also chairman of the Scientific Committee = Cosmetology reported on the activities of this Committee in the = following fields: preservatives, sun tans, hydrochinones and ethical = considerations concerning human trials. The discussions on the latter = aspect should eventually lead to the establishment of standards in human = trials.

III.3. Reporting by other Commission services on matters related = to consumer health.

Dr.E.Anklam of the Food & Drug Analysis / Consumer Protection = unit of the Environment Institute of the Joint Research Centre, = reported on the following research, planned to be realised during the = coming months:

• the validation study of methods for identifying bovine DNA in animal = feeds;
• verification of the temperatures which had been applied when = producing animal meal. This permits to determine whether animal meal had = been treated according to the "133=B0C / 20' / 3 bar" = regulation.
A short note was handed out to all participants. The MDSC/SSC raised = the question whether this research would also cover the ELISA test = currently used in the UK. Dr.Anklam confirmed that she intended to = verify this and that she would contact UK researchers on this matter. As = this research is of high interest for its activities, the MDSC/SSC = expressed the wish to be kept informed about the results of this = research. Mrs.Anklam will take the initiative to contact the SSC, once = research results are available.

Dr.Rohte, Directorate General III - Industry, reported on 2 = scientific co-operation projects which were presently being implemented = under Directive 93/5. One of them concerns the establishment of an = inventory of the data available in Member States on allergens in food = and on their authorisation; the second project covers the establishment = of an inventory of actual knowledge in the Member States on the state of = bacteriological contamination in different types of food.

Mrs.Peutz reported on activities of the Directorate General = VI-Agriculture related to BSE. The items presented were: the Meat = and Bone Meal Decision which had been voted on 8.10.97 with a qualified = majority by the Standing Veterinary Committee; the need for reinforcing = veterinary controls, not only in the E.U., but also at the level of the = whole of continental Europe; several aspects of the UK ban (gelatine, = pet food, life saving drugs, semen, tallow derivatives, ....). Mrs.Peutz = further informed the MDSC that DGVI was working on a proposal for TSE = surveillance at the level of the European Union.

Closing of the meeting

In his closing remarks, Dr.H.Reichenbach, Director General of = DGXXIV warmly thanked Prof.F.Kemper for his more then one year = chairmanship of the Multidisciplinary Scientific Committee and for the = support and all personal efforts put into the Committee. The task had = not been easy, as the MDSC was created in the middle of a European-wide = crisis and as its work was watched closely by all parts of the society. = During 13 meetings, Prof.Kemper succeeded to guarantee the objectivity = and excellency of the Committee in a field where, indeed, very little is = still known.

Prof. Kemper thanked the European Commission and its services not = only for all the support it provided, but also for the good working = atmosphere. He expressed his wish that this would go on in the new = Scientific Steering Committee.

Prof. Pascal then thanked Prof. Kemper on behalf of all the members = of the MDSC, whose critical sense and deep concerns for consumer health = did result in fruitful work, but not always in easy meetings.

The chairman closed the meeting at 17h45

Annex 1: List of participants in the MDSC/SSC meeting of = 16.10.97:

Members of the MDSC/SSC:

Prof.G.Del Real

Prof.M.J.Gibney

Prof.Ph.James

Prof.J.M.Jouany

Prof.F.H.Kemper, chairperson

Prof.W.Klein

Prof.R.Kroes

Prof. G.Pascal

Prof.V.Silano

Prof.M.Vanbelle

Prof.R.Wennig

Prof. P.Willeberg,

Apologies were received from: Prof. D.Dormont, Prof.M.Pocchiari, = Prof.G.Vicari and Prof.M.Wierup

Experts:

Prof.A.Somogyi

Participants from the Commission:

O.Rohte, L.Bansil, E.Zbinden DG III

L.Chambaud, DG V

P.Colombo, I.Peutz, H.Winter DG VI

L.Matthiessen, DG XII

A.Matton DG XV

E.Anklam DG-CCR

DG XXIV: H.Reichenbach, B.Carsin, P.Brunko, J.Costa-David, C.Deckart, = M.Dekindt, J.Kreysa, G.Morrison, W.Penning, J.Moynagh, J.J.Rateau, = J.Savio, S. van de Louw, A.Van Elst, R.Vanhoorde, B.Verleysen, = P.Vossen;

Annex 2:

Agenda of the Meeting of 16 October 1997 of the Multidisciplinary = Scientific Committee/Scientific Steering Committee (MDSC/SSC)

____________

1. Welcome, apologies, introductory remarks

2. Approval of the agenda

3. Approval of the minutes of the meeting of 8 September 1997

Part I: Matters related to BSE

I.1. Specified Risk Materials: further discussion and = possibly adoption of the proposed opinion which was redrafted on the = basis of the discussions of 8 September (This point includes the = discussion on the question whether the intestine should be considered to = be a high risk tissue).

I.2. Meat and Bone Meal: discussion and possibly adoption of an = opinion on the minimum production process conditions required for = inactivating / eliminating the BSE infective agent.

I.3. Tallow and tallow derivatives: examination of newly received = information and requests for clarification on the opinion adopted on = 8.09.97.

I.4. Maternal transmission of BSE: additional questions on the follow = up of the opinion of 8.09.97; planning and approach to prepare an = opinion on these questions.

I.5. Review of the situation concerning the following products listed = in Annex 1 to the Commission Decision 95/362/EC of 11 June 1996:

1. Di-calcium phosphate: progress report (Prof.D.Dormont, = rapporteur);
2. Amino acids and peptides produced from animal origin and derived = products: progress report (Prof.G.Pascal, rapporteur);
I.6. Information on the Export Certified Herd Scheme and on the BSE = status of Australia, New-Zealand, Norway and the USA: reporting by the = Scientific Veterinary Committee (Prof. Del Real).

I.7. Possible link between Bovine Spongiform Encephalopathy and the = new variant Creutzfeldt-Jakob Disease (nvCJD): brief discussion.

PART II: Any other business

II.1. Dates of the next meetings

II.2. Miscellaneous

PART III: Matters related to co-ordination

III.1. Establishment of the 8 new Scientific Committees: the = state of affairs;

III.2. Reports of the chairs of the at present existing scientific = committees;

III.3. Reporting by other Commission services on matters related to = consumer health, including a report by the Joint Research Centre on the = validation study of methods for identifying animal proteins in MBM and = for verifying the temperatures which were applied when producing the = MBM.

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