Original combination of real time PCR and NIRM for the detection and the speciation of animal particles

  • Fumière, O. , Marien, A. , Berben, G. , Baeten, V. & Fernández Pierna, J.A. (2007). Original combination of real time PCR and NIRM for the detection and the speciation of animal particles. CRA-W. Proceedings in: FEED SAFETY International Conference 2007: Methods and Challenges, Namur - Belgique, 27-28/11/2007, 66-67.
Type Conference Proceedings
Year of conference 2007
Title Original combination of real time PCR and NIRM for the detection and the speciation of animal particles
Editor CRA-W
Conference name FEED SAFETY International Conference 2007: Methods and Challenges
Conference location Namur - Belgique
Recnumber 1098
Pages 66-67
Label U15-1098
conference Date 27-28/11/2007
Endnote Keywords PAPs|feed ban|BSE|real-time|NIRM|
Abstract The bovine outbreak of spongiform encephalopathy (BSE) has led the European Union to take several decisions in order to avoid the transmission through the food chain of its causal agent. The Regulation (EC) 999/2001 prohibits explicitly the feeding of mammalian processed animal proteins (PAPs) to ruminants. A temporary MBM ban for all farmed animals (to avoid intra-species recycling) was established in 2001 and changed into a permanent MBM ban by amending the annex of Regulation (EC) 999/2001 through Regulation (EC) 1234/2003. The animal by-product (ABP) regulation EC 1774/2002 prohibits feeding of animals with proteins from the same species and established three categories of ABP's which reflect different levels of food safety including the risk due to Transmissible Spongiform Encephalopathy (TSE). Presently, classical optical microscopy is the only official method in the European Union used to detect constituents of animal origin in compound feeds or in their ingredients. Successfully validated, the method is nevertheless limited to the differentiation of large taxonomic groups i.e. terrestrial animal or fish. There is a tremendous need for techniques enabling to detect more routinely these highly processed animal by-products throughout the feed chain but also to identify their animal origin at the species level. Among the alternative techniques developed for the detection of meat and bone meals, near-infrared microscopy (NIRM) and real time PCR have been used by CRA-W to develop and validate original solutions. In the framework of the Belgian research project FARIMAL (contract nr S-6168) both methods were combined in an original procedure to develop authenticated species specific spectral databases. One thousand and one hundred (1100) particles coming from 24 pure species animal meals (6 cattle, 1 sheep, 7 pig, 2 chicken and 8 fish meals respectively) were analysed by both techniques (PCR was just used to check the species origin of each particle) and discriminant models were constructed in order to predict with its spectral characteristics the species origin of a particle. Based on these libraries, the first results show that 80 % of the cattle particles and 90 % of the fish particles can be correctly assigned by NIRM to their species of origin. It should be stressed that, the NIRM prediction can be confirmed afterwards by real time PCR thanks to a particular protocol allowing the realisation of 5 PCR with the DNA extracted from a single particle.
Author address Fumière Olivier, Quality Department of Agro-food Products, Walloon Agricultural Research Centre (CRA-W), Chaussée de Namur, 24, B-5030 Gembloux, fumiere@cra.wallonie.be
Caption U15-1098-fumiere-2007.pdf
Lien http://safeedpap.feedsafety.org/fs2007/posters/fumiere.php
Authors Fumière, O., Marien, A., Berben, G., Baeten, V., Fernández Pierna, J.A.

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