The molecular composition of milk is influenced by various genetic and environmental factors. Time is one important factor, and the fact that certain milk components change over the course of lactation is widely accepted. Untargeted global metabolomics is an approach to study hundreds of low molecular weight compounds simultaneously. In this study, mass spectrometry-based global metabolomics was used to follow the course of changes in milk (n=133) and blood plasma (n=133) during the early stage of lactation. Little correlation was found between the molecular composition of blood plasma and milk. Blood showed a higher dependence on animal individuality than did milk, in which common evolutions in time resolved. Citrate and lactose had the greatest effect on these changes; however, the most significant changes in milk during the first months of lactation were associated with phosphorylated saccharide levels, whereas the most significant changes in blood plasma were associated with levels of polyunsaturated fatty acids containing phosphatidylcholine. In conclusion, a new systemic approach was used to search for minor metabolites whose concentrations were significantly altered in milk and blood during the first months of lactation.
Adipose tissue plays an important role in a cow's ability to adapt to the metabolic demands of lactation, because of its central involvement in energy metabolism and immunity. High adiposity and adipose tissue resistance to insulin are associated with excessive lipid mobilization. We hypothesized that the response to a glucose challenge differs between cows of different body condition 21 d before and after calving and that the responses are explainable by gene expression in subcutaneous adipose tissue (SAT). In addition, we aimed to investigate insulin resistance with gene expression in SAT and lipid mobilization around parturition. Multiparous Holstein cows were grouped according to body conditions score (BCS) 4 wk before calving, as follows: BCS ≤ 3.0 = thin (T, n = 14); BCS 3.25 to 3.5 = optimal (O, n = 14); BCS ≥ 3.75 = over-conditioned (OC, n = 14). We collected SAT on d -21 and d 21 relative to calving. A reverse-transcriptase quantitative (RT-q)PCR was used to measure gene expression related to lipid metabolism. One hour after the collection of adipose tissue, an intravenous glucose tolerance test was carried out, with administration of 0.15 g of glucose per kg of body weight (with a 40% glucose solution). Once weekly from the first week before calving to the third week after calving, a blood sample was taken. The transition to lactation was associated with intensified release of energy stored in adipose tissue, a decrease in the lipogenic genes lipoprotein lipase (LPL) and diacylglycerol O-acyltransferase 2 (DGAT2), and an increase in the lipolytic gene hormone-sensitive lipase (LIPE). On d -21, compared with T cows, OC cows had lower mRNA abundance of LPL and DGAT2, and the latency of fatty acid response after glucose infusion was also longer (8.5 vs. 23.3 min) in OC cows. Cows with higher insulin area under the curve on d -21 had concurrently lower LPL and DGAT2 gene expression and greater concentration of fatty acids on d -7, d 7, and d 14. In conclusion, high adiposity prepartum lowers the whole-body lipid metabolism response to insulin and causes reduced expression of lipogenic genes in SAT 3 weeks before calving. In addition, more pronounced insulin release after glucose infusion on d -21 is related to higher lipid mobilization around calving, indicating an insulin-resistant state, and is associated with lower expression of lipogenic genes in SAT.
Milk composition has been known to change during lactation. To help understand the changes in metabolic profile throughout the whole lactation, liquid chromatography mass-spectrometry was used to analyze 306 milk samples from 82 primi- and multiparous dairy cows. Changes in metabolic profile common to all cows throughout lactation were ascertained based on principal component and general linear model analysis. Sets of specific markers; for instance, 225, 397, and 641-642 m/z (positive mode), and 186, 241, and 601-604 (negative mode), with at least a 1.5-fold higher intensity during the first 60 d compared with the last 60 d of lactation were observed. The metabolome was affected by parity and milking time. Markers, identified as peptides differentiating parity, were observed. A significant increase for citrate was observed in evening milk. Milk coagulation traits were strongly animal specific. The curd firmness values were influenced by milking time. Sets of markers were associated with curd firmness in positive (197 m/z) and negative (612, 737, 835, 836, 902, 1000, 1038, and 1079 m/z) ion mode.