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Worldwide growth and performance-enhancing substances are used in cattle husbandry to increase productivity. In certain countries however e.g., in the EU, these practices are forbidden to prevent the consumers from potential health risks of substance residues in food. To maximize economic profit, ‘black sheep‘ among farmers might circumvent the detection methods used in routine controls, which highlights the need for an innovative and reliable detection method. Transcriptomics is a promising new approach in the discovery of veterinary medicine biomarkers and also a missing puzzle piece, as up to date, metabolomics and proteomics are paramount. Due to increased stability and easy sampling, circulating extracellular small RNAs (smexRNAs) in bovine plasma were small RNA-sequenced and their potential to serve as biomarker candidates was evaluated using multivariate data analysis tools.After running the data evaluation pipeline, the proportion of miRNAs (microRNAs) and piRNAs (PIWI-interacting small non-coding RNAs) on the total sequenced reads was calculated. Additionally, top 10 signatures were compared which revealed that the readcount data sets were highly affected by the most abundant miRNA and piRNA profiles. To evaluate the discriminative power of multivariate data analyses to identify animals after veterinary drug application on the basis of smexRNAs, OPLS-DA was performed. In summary, the quality of miRNA models using all mapped reads for both treatment groups (animals treated with steroid hormones or the β-agonist clenbuterol) is predominant to those generated with combined data sets or piRNAs alone. Using multivariate projection methodologies like OPLS-DA have proven the best potential to generate discriminative miRNA models, supported by small RNA-Seq data. Based on the presented comparative OPLS-DA, miRNAs are the favorable smexRNA biomarker candidates in the research field of veterinary drug abuse.

Veterinary drug abuse is a common committed crime in food-producing animal husbandry with far-reaching impact on animal´s health, human nutrition and thereby competitive sports. To prevent consumers from possible health risks by consumption of contaminated meat, a reliable and fraud-resistant surveillance strategy need to be established. The potential of the circulating small RNA transcriptome as innovative source of biomarker candidates was evaluated using the state-of-the-art holistic fingerprinting technology called Next-Generation Sequencing (NGS). In der Fleischproduktion werden in vielen Ländern der Erde hormonelle und pharmazeutische Wachstums- und Leistungsförderer appliziert. In der Europäischen Union ist es verboten anabole Substanzen in der Rindermast einzusetzen. Dadurch werden Konsumenten vor potentiell gesundheitsschädlichen Rückständen im Fleisch geschützt und unabsichtliches Doping im Leistungssport über den Verzehr von kontaminiertem Fleisch verhindert. Um den illegalen Einsatz von anabolen Agenzien zuverlässig und betrugssicher nachweisen zu können, ist ein neuer und innovativer Weg zu beschreiten. Mithilfe von molekularem Profiling, dem Next-Generation Sequencing (NGS), können Genexpressionsveränderungen auf der Transkriptomebene höchst sensibel detektiert werden. So konnte das Potenzial des small RNA Transkriptoms validiert und neue Biomarker-Kandidaten gefunden werden.