Meelis Ots

Alterations in milk and blood metabolomes during the first months of lactation in dairy cows.

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.

Alterations in milk metabolome and coagulation ability during the lactation of dairy cows

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.

CRUDE GLYCEROL AS GLYCOGENIC PRECURSOR IN FEED, EFFECTS ON MILK COAGULATION PROPERTIES AND METABOLIC PROFILES OF DAIRY COWS

As grain prices rise, the search for alternative glycogenic precursors in animal feed becomes increasingly important, and this study was conducted to determine if the replacement of starch with glycerol, as an alternative glycogenic precursor, affects the milk metabolic profile and milk coagulation ability, and therefore the quality of the milk. Eight primiparous mid-lactation Holstein cows were fed during a replicated 4 × 4 Latin square trial with four different isoenergetic rations: (1) control (T0) fed a total mixed ration (TMR) with barley meal; (2) group T1, decreased barley content, replaced isoenergetically with 1 kg crude glycerol; (3) group T2, the barley meal was replaced with 2 kg of crude glycerol; and (4) group T3 the barley meal was replaced with 3 kg of crude glycerol. Rumen, blood and milk samples were collected at the end of every 21-d treatment period. Rumen samples were analysed for proportion of total volatile fatty acid (VFA), blood samples for insulin and glucose, and milk for metabolites (e.g. citric-acid cycle compounds). The change in glycogenic precursors had a positive effect on rumen VFA proportions; the proportion of propionic acid increased (P < 0.001). Milk protein (P < 0.001) and curd firmness (P < 0.001) both increased. The increase in milk protein concentration may have been due to an increase in microbial protein. Regarding the milk metabolic profiles, different signals were positively associated with coagulation ability and change in the diet. Based on this study, changing the glycogenic precursor in animal diet in this way is possible, and may have no immediate deleterious consequences on milk quality or cow health. Indeed, there is evidence for benefits from this substitution.

Trans Fatty Acid Contents in Selected Dietary Fats in the Estonian Market

UNLABELLED In response to public concern, this study assessed the fatty acid (FA) composition of blended spreads, margarines and shortenings in the Estonian retail market in 2011. Special attention was paid to the trans fatty acids (TFA) composition. The changes in these characteristics of selected dietary fats in the market over recent years are also presented. Twenty-six edible fat brands, available in the Estonian retail market in 2011, were purchased and FA compositions were analyzed by chromatography. Saturated fatty acids (SFA) were the dominant group of FAs for all blended spreads (49.6 to 65.8%), and for the majority of shortenings (from 21.1 to 54.6%). Cis monounsaturated fatty acids (MUFA) were the dominant group of FAs for the majority of margarines, ranging from 25.3% to 50.5%. The total TFA for blended spreads varied from 1.18% to 9.08%, for margarines from 0.04% to 34.96% and for shortenings from 0.14% to 39.50%. Octadecenoic (C18:1) isomers were the main TFA found. Compared to 2008/2009, the industrially produced TFA (IP-TFA) content in several of the dietary fat brands was much reduced in 2011. This voluntary reformulation was probably a response to consumer demand associated with a public health campaign directed against IP-TFA in Estonian foods, and were mainly achieved by replacing TFA with SFA C12:0-C16:0. PRACTICAL APPLICATION Present paper is directed toward public health related institutions and food industries producing foods with potentially high contents of trans fatty acids (TFA). According to the public concern TFA content in domestic blended spreads has declined significantly over the past 3 y in Estonia. The reduction in the TFA content was achieved by replacing TFA with saturated fatty acids (SFA) (C12:0-C16:0). To shift food composition toward healthier product formulations, mandatory labeling of the sum of IP-TFA and SFA (C12:0-C16:0) was recommended.

Adipose tissue insulin receptor and glucose transporter 4 expression, and blood glucose and insulin responses during glucose tolerance tests in transition Holstein cows with different body condition

Glucose uptake in tissues is mediated by insulin receptor (INSR) and glucose transporter 4 (GLUT4). The aim of this study was to examine the effect of body condition during the dry period on adipose tissue mRNA and protein expression of INSR and GLUT4, and on the dynamics of glucose and insulin following the i.v. glucose tolerance test in Holstein cows 21 d before (d -21) and after (d 21) calving. Cows were grouped as body condition score (BCS) ≤3.0 (thin, T; n = 14), BCS = 3.25 to 3.5 (optimal, O; n = 14), and BCS ≥3.75 (overconditioned, OC; n = 14). Blood was analyzed for glucose, insulin, fatty acids, and β-hydroxybutyrate concentrations. Adipose tissue was analyzed for INSR and GLUT4 mRNA and protein concentrations. During the glucose tolerance test 0.15 g/kg of body weight glucose was infused; blood was collected at -5, 5, 10, 20, 30, 40, 50, and 60 min, and analyzed for glucose and insulin. On d -21 the area under the curve (AUC) of glucose was smallest in group T (1,512 ± 33.9 mg/dL × min) and largest in group OC (1,783 ± 33.9 mg/dL × min), and different between all groups. Basal insulin on d -21 was lowest in group T (13.9 ± 2.32 µU/mL), which was different from group OC (24.9 ± 2.32 µU/mL. On d -21 the smallest AUC 5-60 of insulin in group T (5,308 ± 1,214 µU/mL × min) differed from the largest AUC in group OC (10,867 ± 1,215 µU/mL × min). Time to reach basal concentration of insulin in group OC (113 ± 14.1 min) was longer compared with group T (45 ± 14.1). The INSR mRNA abundance on d 21 was higher compared with d -21 in groups T (d -21: 3.3 ± 0.44; d 21: 5.9 ± 0.44) and O (d -21: 3.7 ± 0.45; d 21: 4.7 ± 0.45). The extent of INSR protein expression on d -21 was highest in group T (7.3 ± 0.74 ng/mL), differing from group O (4.6 ± 0.73 ng/mL), which had the lowest expression. The amount of GLUT4 protein on d -21 was lowest in group OC (1.2 ± 0.14 ng/mL), different from group O (1.8 ± 0.14 ng/mL), which had the highest amount, and from group T (1.5 ± 0.14 ng/mL). From d -21 to 21, a decrease occurred in the GLUT4 protein levels in both groups T (d -21: 1.5 ± 0.14 ng/mL; d 21: 0.8 ± 0.14 ng/mL) and O (d -21: 1.8 ± 0.14 ng/mL; d 21: 0.8 ± 0.14 ng/mL). These results demonstrate that in obese cows adipose tissue insulin resistance develops prepartum and is related to reduced GLUT4 protein synthesis. Regarding glucose metabolism, body condition did not affect adipose tissue insulin resistance postpartum.