Serum biochemistry analytes were analysed with the Olympus AU400 analyser with following techniques and OSR reagents (Olympus System Reagent)35 (link),36 (link): glucose (HK method), urea (GLDH method), creatinine (Jaffé method), cholesterol (CHOP-PAP-method), triglyceride (GPO-PAP method), total protein (Biuret method), ALT (International Federation of Clinical Chemistry (IFCC) method), AST (IFCC method) and GGT (IFCC method). NEFAs were determined with NEFA-C reagent (Wako Chemicals GmbH, Neuss, Germany). Quality control protocols were based on the daily quantification of two control sera of low and high concentration (Control serum I and Control serum II, Beckman Coulter) Haptoglobin was determined by a colorimetric method (Tridelta, Ireland). Insulin and IGF-I were determined by ELISA (Bovine Insulin ELISA from Mercodia, Uppsala, Sweden and Human IGF-I ELISA E20 from Mediagnost, Reutlingen, Germany). This reagent is suitable for bovine samples, as declared by the manufacturer, but the cross-reactivity was not assessed by the authors.
Intra-assay Coefficients of Variation (CV) were: glucose (1.7%), urea (2.1%), creatinine (1.4%), cholesterol (1.1%), triglycerides (3.0%), total protein (0.9), ALT (1.7%), AST (1.0%), GGT (0.6%), NEFAs (2.7%), Haptoglobin (4.1%), insulin (4.1%) and IGF-1 (4.4%).
Intra-assay Coefficients of Variation (CV) were: glucose (1.7%), urea (2.1%), creatinine (1.4%), cholesterol (1.1%), triglycerides (3.0%), total protein (0.9), ALT (1.7%), AST (1.0%), GGT (0.6%), NEFAs (2.7%), Haptoglobin (4.1%), insulin (4.1%) and IGF-1 (4.4%).