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Proteomic identification of plasma proteins as markers of growth promoter abuse in cattle

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Abstract

Growth-promoting agents are continually misused for increasing animal growth and fraudulent gain in the meat industry, yet detection rates from conventional targeted testing for drug residues do not reflect this. This is because testing currently relies on direct detection of drugs or related metabolites and administrators of such compounds can take adaptive measures to avoid detection through the use of endogenous or unknown drugs, and low dose or combined mixtures. New detection methods are needed which focus on the screening of biological responses of an animal to such growth-promoting agents as it has been demonstrated that genomic, proteomic and metabolomics profiles are altered by xenobiotic intake. Therefore, an untargeted proteomics approach using comparative two-dimensional gel electrophoresis (2DE) was carried out to identify putative proteins altered in plasma after treatment with oestradiol, dexamethasone or prednisolone. Twenty-four male cattle were randomly assigned to four groups (n = 6) for experimental treatment over 40 days, namely a control group of non-treated cattle, and three groups administered 17β-oestradiol-3-benzoate (0.01 mg/kg, intramuscular), dexamethasone sodium phosphate (0.7 mg/day, per os) or prednisolone acetate (15 mg/day, per os), respectively. Plasma collected from each animal at day 25 post study initiation was subjected to proteomic analysis by 2DE for comparison of protein expression between treated and untreated animals. Analysis of acquired gel images revealed 22 plasma proteins which differed in expression by more than 50 % (p < 0.05) in treated animals compared to untreated animals. Proteins of interest underwent identification by LC–MS/MS analysis and were found to have associated roles in transport, blood coagulation, immune response and metabolism pathways. In this way, seven proteins are highlighted as novel biomarker candidates including transthyretin which is shown to be significantly increased in all treatment groups compared to control animals and potentially may find use as global markers of suspect anabolic practice.

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Acknowledgments

This work was financially supported by the UK Government Department for Employment and Learning, Northern Ireland (DELNI) through the Advanced ASSET centre with no conflict of interest imposed and all research was conducted under good laboratory practice.

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Authors and Affiliations

  1. Institute for Global Food Security, School of Biological Sciences, Queen’s University Belfast, 18-30 Malone Road, Belfast, Co. Antrim, BT9 5BN, Northern Ireland

    Ruth A. Kinkead, Christopher T. Elliott & Mark H. Mooney

  2. Department of Animal Pathology, University of Turin, Via L. Da Vinci 44, 10095, Grugliasco, TO, Italy

    Francesca T. Cannizzo & Bartolomeo Biolatti

Authors
  1. Ruth A. Kinkead
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  2. Christopher T. Elliott
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  3. Francesca T. Cannizzo
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  4. Bartolomeo Biolatti
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  5. Mark H. Mooney
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Correspondence to Ruth A. Kinkead.

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Published in the topical collection on Hormone and Veterinary Drug Residue Analysis with guest editors Siska Croubels, Els Daeseleire, Sarah De Saeger, Peter Van Eenoo, and Lynn Vanhaecke.

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Kinkead, R.A., Elliott, C.T., Cannizzo, F.T. et al. Proteomic identification of plasma proteins as markers of growth promoter abuse in cattle. Anal Bioanal Chem 407, 4495–4507 (2015). https://doi.org/10.1007/s00216-015-8651-0

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  • DOI: https://doi.org/10.1007/s00216-015-8651-0

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