Research project CP/58 (Research action CP)
Context
Food safety is a very important issue all around the world. Due to the immense economical implications concerned with global food safety, preventive actions are much needed: the development of a sustainable detection method for use in routine screenings is a first and indispensable step. The screening of food products (and their production sites and processing units) to detect possible pathogenic bacteria is an essential part of a solid preventive policy. Due to the specific characteristics and the high level of variability among bacterial species, the development of functional methods to detect the most important pathogens is both a national and international issue. Developing new detection methods will allow public authorities to compose a new preventive policy in order to improve food safety.
Project description
Objectives
The project aims at developing a detection methodology for all VTEC (verocytotoxin producing Escherichia coli) pathogens, including non-sorbitol fermenting E. coli O157. At this moment, there is no effective method available to detect these hazardous organisms. In Belgium, no detection of these extreme severe pathogens (except typical E. coli O157) is performed in food and farm related samples. Consequently, no information exists about the prevalence and the contamination sources. Nevertheless, human clinical samples collected in Belgium indicate that non-O157 VTEC cause 80% of hospitalised VTEC infections and 23% of the extreme life-threatening forms of the disease. This project aims at consolidating the Belgian scientific potential in this field of research. Besides a detection methodology also primary information about the prevalence of the pathogens and the sampling efficiency at the farm level will be available at the end of the project. These results will form the basis for the decision makers to undertake action for setting up a combative prevention programme. The project generates data which will allow public authorities, represented by an extended users committee, to develop tools that will provide a reliable scientific basis for working out an appropriate strategy for prevention of human infections with pathogenic VTEC.
Methodology
The detection and isolation methodology for non-O157:H7 VTEC will be based on recent available scientific information concerning biochemical characteristics of the human clinical isolates. These biochemical characteristics will be used to improve differentiation from other Enterobacteriaceae and from non-pathogenic E. coli. In this respect the haemolytic property identified on blood agar, the higher acid tolerance and the specific resistance to certain antibiotics and other chemical substances will be applied.
For the most important serotypes (O26, O111, O103, O145) magnetic beads coated with specific antibodies are recently commercially available. These magnetic beads will be used to improve the isolation efficiency of these serotypes. Identification of non-O157:H7 VTEC isolates, pathogenic for human, will be performed by polymerase chain reaction (PCR) based methodology. The identification will be performed on pure cultures picked from colonies on plates and from enrichment cultures. Identification will also be performed on plates by means of colony hybridisation.
The detection and isolation methodology will be evaluated on milk, raw milk cheese, minced beef, Belgian dry sausage and cattle faeces. Naturally and artificially contaminated samples will be applied for this purpose.
Interaction between the different partners
Both partners in this project constantly exchange information: pathogenic and non-pathogenic strains will be centralized by CLO-DVK. Charcterisation of the strains will be performed by both partners at the same time: different tasks and subtasks will be assigned according to the specific fields of experience. Testing of the detection and isolation methods is carried out on food products by both partners in their specific field of experience. CLO-DVK will perform tests on milk, milk-related foodproducts and farmsamples, while the Faculty of Veterinary Sciences will perform tests on slaughterhouse related samples, meat and meatproducts (Belgian dry sausage, carcasses and minced beef).
Expected results
In this project a routine detection and isolation methodology for non-O157 E. coli VTEC and for sorbitol positive O157 in food products and animal related farm samples will be developed. The project will result in an isolation and detection scheme for non-O157 E. coli VTEC and for sorbitol positive O157 strains in food products such as raw milk, raw milk cheese, minced meat and in farm related samples as faeces and overshoes. This project will offer the molecular biological tools for routine identification and characterization of VTEC strains.
During the developmental process primary information about the prevalence in Belgium of different pathogenic VTEC types in critical food products and on the farm will be collected. Primary information will be transferred to the public authorities in order to allow them to take the first steps in an effective plan to prevent human infections. Knowledge of the prevalence of contaminations at farm level and cross contaminations of pathogenic micro-organisms present in foods, like VTEC, is of strategic importance to prevent human infections.
The project will give indications for the most effective sampling plan for determination of farm status concerning contamination with pathogenic VTEC. This primary information will form the basis for the further development of a decent combative programme at farm level. This programme will decrease the contamination pressure in the primary processing plants and will prevent further spread of the pathogenic strains to other farms and in the environment. The detection methodology for non-O157 VTEC and its availability in Belgium will form the first and necessary step in a programme for prevention of human infection due to contaminated food products.
Partners
Activities
In order to reach the goals of this project, a multidisciplinary team of two different scientific institutions has been composed. Coördinator of this team, Prof. L. De Zutter, is staff member of The University of Ghent (Faculty of Veterinary Sciences). He is mainly specialised in detection methods for tracing pathogens in meat and meat products.
As a partner of the multidisciplinary team, the research group led by Dr. L. Herman and Dr. M. Heyndrickx (CLO-DVK) has plenty of experience in detecting bacteria in milk and milk related products, with the emphasis on molecular detection and identification methods.
Contact information
Co-ordinator
Lieven De Zutter
Universiteit Gent (UGent)
Faculteit Diergeneeskunde - Laboratorium voor Hygiëne en Technologie
Salisburylaan 133
B-9820 Merelbeke
Tel: +32 (0)9 264 74 55
Fax: +32 (0)9 264 74 91
lieven.dezutter@ugent.be
http://allserv.ugent.be/~hdbraban/
Partners
Lieve Herman
Ministerie van de Vlaamse Gemeenschap
Centrum voor Landbouwkundig Onderzoek - Gent (CLO) - Departement voor de Kwaliteit van Dierlijke Producten en Transformatietechnologie (DVK)
Brusselsesteenweg 370
B-9090 Melle
Tel: +32 (0)9 272 30 10
Fax: +32 (0)9 272 30 01
l.herman@clo.fgov.be
http://www.clo.fgov.be/dvk/
Marc Heyndrickx
Ministerie van de Vlaamse Gemeenschap
Centrum voor Landbouwkundig Onderzoek - Gent (CLO) - Departement voor de Kwaliteit van Dierlijke Producten en Transformatietechnologie (DVK)
Brusselsesteenweg 370
B-9090 Melle
Tel: +32 (0)9 272 30 44
Fax: +32 (0)9 272 30 01
m.heyndrickx@clo.fgov.be
http://www.clo.fgov.be/dvk/
Users committee
Pr Dr Denis Piérard
Hôpital universitaire, Vrije Universiteit Brussel
Laboratoire de microbiologie
Dr H. Imberechts
Centre de recherche vétérinaire et agrochimique
Département de bactériologie et d'immunologie
Pr Dr Johan Debevere
Laboratoire de microbiologie alimentaire et de conservation des aliments
Département de technologie alimentaire et de nutrition
Faculté des sciences agricoles et biologiques appliquées
Université de Gand
Pr Georges Daube
Faculté de médecine vétérinaire
Université de Liège
Pr Dr Chris Michiels
Laboratoire de microbiologie alimentaire
Département de technologie alimentaire et microbienne
Faculté des sciences agricoles et biologiques appliquées
Katholieke Universiteit Leuven
Dr Katelijne Dierick, DVM
Institut scientifique de la santé publique - Louis Pasteur
Section alimentaire
M. Eddy Leloup
Boerenbond – service d'étude
Dr Vincent Merken
SPF Economie – administration qualité et sécurité
M. Verhaeghe Alain
Fédération belge des entreprises de distribution (Fedis)
Dr Marie-Thérèse Parmentier
NVS-Boviqual
Mme Sigrid Lauryssen
Test Achats / Test Aankoop
Development of detection method for non-0157-pathogenic-E. coli in food: first step in prevention : final report
De Zutter, Lieven - Possé, Björn - Herman, Lieve ... et al Brussels : Belgian Science Policy, 2006 (SP1641)
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