Entwicklung einer PCR-Methode zum tierartspezifischen Nachweis des Erhitzungsgrades bei Futtermitteln tierischer Herkunft

• Forschungsstelle: BLV
• Projektnummer: 1.00.16
• Projekttitel Englisch: Development of a PCR-Based Test for Species-Specific Determination of Heat Treatment Conditions for Animal Meals as an Effective Prophylactic Method for Bovine Spongiform Encephalopathy

Schlüsselwörter (Deutsch)

BSE, ELISA, PCR, Tiermehl

Schlüsselwörter (Englisch)

BSE, ELISA, PCR, Animal meals

Kurzbeschreibung (Englisch)

To ensure the exclusion of Bovine Spongiform Encephalopathy agent (BSE) and other pathogens from animal feeds the European Union (EU) specifies specific treatment conditions in Decision 96/449 EC (Anon. 1996). As regards imported animal feedstuffs, the Swiss (Anon. 1993) and EU prescriptions (Anon. 1997) provided to rendering companies procedures for heat treatment of animal wastes from slaughterhouses which guarantee the inactivation of all pathogenic materials. In order to guarantee the denaturation of hazardous agents in animal waste products and be considered as BSE safe in particular the meat and bone (MBM) meal manufacturers must heat these products at 133°C for 20 min. at a pressure of 3 bar.
To verify the application of the above legal mentioned prescriptions, the imported and homemade animal feedstuffs have to pass an ELISA based test (Anonymous, 1994). This test involves the detection of non denatured pork proteins in meat meal and MBM and has been evaluated in a collaborative study including 21 laboratories from 12 European countries (Unglaub et al. 1999).
The ultimate aim of the heat treatment conditions is the elimination of possible BSE agents from animal waste materials. One can easily imagine that an animal feedstuff made exclusively out of bovine material and which has not been treated according to the prescribed conditions could pass the pork-based antibody ELISA-test.
Recently a rapid and simple polymerase chain reaction (PCR) method for species specific identification of meat species from cooked meat and meat products has been described by Matsunaga et al. (1999). By means of a simple or multiplex PCR this method is able to specifically detect a targeted species in a mixture of cooked animal meat materials. All these species-specific test methods were focused on meat and meat products for commercial, economic, ethnic or religious reasons and not for prophylactic concerns.
We would like to use this simple and rapid PCR-based method for the verification of an effective heat treatment in order to confirm the inactivation of possible BSE agents from feedingstuffs.

Projektziele (Englisch)

PCR-based test for species-specific determination of heat treatment conditions for animal meals as an effec-tive prophylactic method for bovine spongiform encephalopathy.

Abstract (Englisch)

Both PCR and ELISA assays were able to detect poorly heat-treated feedingstuffs if there was enough pork-based material present. The proposed species-specific PCR test, however, showed a higher sensitivity and specificity as it specifically detected bovine material. Nevertheless, the PCR assay will not detec bovine material in properly heat-treated feeds as the DNA is too fragmented. It is, however, very useful as a rapid, sensitive and sprcific method for the routine screening of animal meals with regard to prophylaxis of BSE.

Publikationen / Ergebnisse (Englisch)

Bin Kingombe, C.I., Lüthi, E., Schlosser, H., Howald, D., Kuhn, M., Jemmi Th. (2001): A PCR-based test for species-specific determination of heat treatment conditions of animal meals as an effective prophylactic method for bovine spongiform encephalopathy. Meat Science 57, 35-41.

Bin Kingombe, C.I., Lüthi, E., Schlosser, H., Howald, D., Kuhn, M., Jemmi Th. (2001): A PCR-based test for species-specific determination of heat treatment conditions of animal meals as an effective prophylactic method for bovine spongiform encephalopathy. Meat Science 57, 35-41.