ABSTRACT: This study was undertaken to identify metabolite differences in plasma of dairy cows with a normal or high calving body condition score (CBCS), using untargeted liquid chromatography-mass spectrometry (LC-MS) metabolomics. Sixteen multiparous dairy cows were assigned to one of two groups based on CBCS (0 to 5 scale): Normal group (NBCS, 3.25 ? BCS ? 3.5, n = 8), and high BCS group (HBCS, BCS ? 4, n = 8). Plasma samples were collected for metabolomics analysis and evaluation of biomarkers of lipid metabolism (nonesterified fatty acid (NEFA) and ?-hydroxybutyrate (BHB)), and cytokines (leptin, adiponectin, tumor necrosis factor-? (TNF-?) and interleukin 6 (IL-6)). A total of 23 differential metabolites were identified, and functional analyses were performed using the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. Among these metabolites, the concentrations of six lysophosphatidylcholines and one phosphatidylethanolamine, were lower in the HBCS group than in the NBCS group (p < 0.01). Furthermore, these metabolites were involved in these four pathways, among others: glycerophospholipid metabolism, retrograde endocannabinoid signaling, autophagy, and glycosylphosphatidylinositol (GPI)-anchor biosynthesis (p < 0.05). In addition, plasma concentrations of leptin (p = 0.06) and TNF-? (p = 0.08) tended to be greater while adiponectin (p = 0.09) lower in HBCS cows than in NBCS cows. The concentrations of NEFA, BHB, or IL-6 did not differ between NBCS and HBCS groups. More importantly, based on the results of the Spearman's correlation analysis, the seven important metabolites were negatively correlated with indices of lipid metabolisms, proinflammatory cytokines, and leptin, but positively correlated with adiponectin. These results demonstrate that CBCS has a measurable impact on the plasma metabolic profile, even when NEFA and BHB are not different. In addition, the identified differential metabolites were significantly correlated to lipid metabolism and inflammation in the over-conditioned fresh cows, which are expected to render a metabolic basis for the diseases associated with over-conditioned dry cows.
