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Developement of an enzyme-linked assay (ELISA) for determination of pyrrolizidine alkaloid (PA)-protein adducts as biomarkers for the detection of PA-exposure in livestock
Project
Project code: MRI-PBE-08-1010 ELISA
Contract period: 01.01.2021
- 31.12.2023
Purpose of research: Experimental development
The project is part of the superordinate research project Pyrrolizidine Alkaloids (PA) Safe Feed. PAs are a group of phytotoxins widely present in herbal plants. Many PA-containing plants can cause severe acute and chronic liver injury in both, humans and animals.
Based on new toxicological risk assessments, plants, seeds or parts of plants containing pyrrolizidine alkaloids (PA) have been classified as undesired substances in food and feed by the European Food Safety Authority (EFSA) and some national authorities, including the German Federal Institute for Risk Assessment (BfR). Due to the increase of greening measures and/or extensive cultivation practices in recent years, together with the increasing temporary use of verges and fallow lands for fodder production due to dry summers, there is an increased risk that plants containing PAs, e.g. Senecio vulgaris or Senecio jacobaea may enter the feed chain of livestock by grazing or via hay/silage. This increases the risk of poisoning of food-producing farm animals. To confirm PA-exposure in farm animals, it would be important to have a reliable PA-biomarker. Ideally, a biomarker would not only be useful for an early diagnosis of toxicologically relevant PA-exposure of farm animals, but would also offer the additional possibility to guarantee the quality and safety of food of animal origin with PA-exposure by defining for example a waiting period after exposure. The aim of this project is to develop a specific and sensitive ELISA assay for practical application to detect PA-(pyrrole)-protein adducts in blood samples of farm animals.
The project runs in cooperation with the Technical University Carolo-Wilhelma of Braunschweig. The MRI will test the monoclonal and polyclonal antibodies produced by an external company for their specificity and binding properties in the sandwich ELISA or the competitive immunassay. In further steps, a pyrrolizidine alkaloid liver intoxication rodent model is used to determine whether the antibodies produced / identified (from the previously established ELISA) are able to detect PA-protein adducts in the blood of animals exposed to PA. In addition to the detection of PA-protein adducts, it should also be checked whether the amount of PA-protein adducts formed in the blood correlates with the amount of adducts in the liver and with the extent of the hepatotoxicity. The in-vitro and in-vivo data obtained during the new development and validation of the pyrrolizidine alkaloid ELISA system should used in a second step for its suitability for practical use. Samples are used which have already been obtained from the already started project: PA-SAFE-FEED: Studien zum Transfer von Pyrrolizidinalkaloiden in Nutztieren (Verbundvorhaben), prepared and preserved for this purpose. These samples from dairy cows, dairy sheep and dairy goats, which have received a defined and traceable PA diet, allow a qualification/evaluation of the new ELISA test system under practical conditions.
Section overview
Subjects
- Animal health
- Toxicology
Framework programme
Funding programme
Excutive institution
MRI - Department of Physiology and Biochemistry of Nutrition (MRI-PBE)
