Merike Henno

A comparison of the methods for determination of the rennet coagulation properties of milk

Abstract The objective of the present study was to compare two methods for measuring the rennet coagulation properties of milk: the Formagraph, and the Optigraph. The parameters used to describe rennet coagulation of 81 milk samples were rennet coagulation time (RCT, Formagraph, and R, Optigraph) and curd firmness 30 min after the addition of rennet (E30, Formagraph, and A30, Optigraph). New Optigraph system recipes were found: R slope = 1.784 and R offset = − 2.303. Using the above Optigraph system recipes, no statistically significant differences were found in rennet coagulation time measurements in either of the studied methods. Regarding curd firmness, the best approximation to describe the relationship between the methods was polynomial: E30= − 0.0357*A30 2+2.8795*A30 −5.2991 (R 2 =0.925). Such strong correlations indicate that the two techniques for detecting milk coagulation properties are comparable and that the data obtained by these methods allow for data interpretation in studies using either method.

Comparison of the metabolic profiles of noncoagulating and coagulating bovine milk

We hypothesize that, through milk composition and different milk metabolites, it is possible to characterize the technological properties (e.g., coagulation) of milk. In this research, liquid chromatography mass spectrometry was used to obtain profiles of low molecular weight organic compounds in 143 milk samples. The metabolic profiles of milk from cows were correlated with their coagulation properties. Using multivariate data analysis methods, we demonstrated that the metabolic profiles of the milk were correlated with coagulation ability. Several marker ions responsible for differential coagulation were found. Although not all affected metabolites could be identified, the most significant differences were found for carnitine and oligosaccharides. Exploitation of these results may increase the use of biomarkers to assess the coagulation ability of milk. This study represents the first large-scale metabolomic profiling of noncoagulating and coagulating bovine milk samples in Estonia.

Relationships between milk coagulation property traits analyzed with different methodologies.

Milk coagulation properties (MCP) analysis is performed using a wide range of methodologies in different countries and laboratories, using different instruments, coagulant activity in the milk, and type of coagulant. This makes it difficult to compare results and data from different research. The aims of this study were to propose a method for the transformation of values of rennet coagulation time (RCT) and curd firmness (a(30)) and to predict the noncoagulation (NC) probability of milk samples analyzed using different methodologies. Individual milk samples were collected during the morning milking in October 2010 from each of 165 Holstein-Friesian dairy cows in 2 freestall barns in Italy, and sent to 3 laboratories for MCP analysis. For each laboratory, MCP analysis was performed using a different methodology: A, with a computerized renneting meter instrument using 0.051 international milk clotting units (IMCU)/mL of coagulant activity; B, with a Lattodinamografo (Foss-Italia, Padova, Italy) using 0.051 IMCU/mL of coagulant activity; and C, with an Optigraph (Ysebaert, Frépillon, France) using 0.120 IMCU/mL of coagulant activity. The relationships between MCP traits were analyzed with correlation and regression analyses for each pair of methodologies. For each MCP trait, 2 regression models were applied: model 1 was a single regression model, where the dependent and independent variables were the same MCP trait determined by 2 different methodologies; in model 2, both a(30) and RCT were included as independent variables. The NC probabilities for laboratories with the highest number of NC samples were predicted based on the RCT and a(30) values measured in the laboratories with lower number of NC samples using logistic regression and receiver operating characteristic analysis. The percentages of NC samples were 4.2, 11.5, and 0.6% for A, B, and C, respectively. The transformation of MCP traits was more precise with model 1 for RCT (R(2): 0.77-0.82) than for a(30) (R(2): 0.28-0.63). The application of model 2 was needed when the C measurements were transformed into the other scales. The analyses of NC probabilities of milk samples showed that NC samples from one methodology were well distinguishable (with an accuracy of 0.972-0.996) based on the rennet coagulation time measured with the other methodology. A standard definition for MCP traits analysis is needed to enable reliable comparisons between MCP traits recorded in different laboratories and in different animal populations and breeds.

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.

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.