Antioxidant nature of bovine milk beta-lactoglobulin

Heating is necessary for processing milk in the dairy industry, which evidently produces a conformational change in β-lactoglobulin (β-LG). β-Lactoglobulin, a major protein that accounts for approximately 10 to 15% of total milk proteins, is a globular protein consisting of 162 AA with a relative molecular mass of 18.4 kDa. The purpose of the present study was to determine the antioxidant role of β-LG in milk and the possible mechanism involved. We showed that β-LG is a mild antioxidant whose potency is less than that of vitamin E and probucol (the latter being an antioxidant used for clinical therapy). The conversion of the β-LG monomer to dimer was responsible, in part, for the mode of action in protecting low-density lipoproteins against copper-induced oxidation. Cross-linking the free thiol groups of β-LG by heating (100°C for 2 min), or chemically modifying the β-LG by carboxymethylation to block the thiol groups resulted in a substantial loss of antioxidant activity. The data suggest that Cys-121 plays an essential role in the antioxidant nature of β-LG. By using an anti-LG antibody affinity column to deplete the β-LG from milk, we observed from the lost antioxidant activity that β-LG contributes approximately 50% of the total activity. Because β-LG is extremely sensitive to thermal denaturation, to maintain its antioxidant nature, dairy products consumed daily should not be overheated in order to maintain its antioxidant nature.     

Effect of kappa-casein polymorphism on milk composition in the Orobica goat

The aim of this work was to study the effects of isoelectrofocusing (IEF) milk protein variants on milk composition in the Italian Orobica goat breed, which is characterized by a rather high frequency of the κ-casein (CSN3) B^IEF allele. Significant associations were found between the IEF phenotype and protein and casein percentages. A favorable effect of the CSN3 B^IEF variant was found for both protein and casein percentages, with a codominance trend for the 3 phenotypes: BB > AB > AA. Depending on the selection purpose, emphasis could be given to different κ-casein variants in breeding. The high frequency of B^IEF could be exploited in breeding strategies to improve the protein and casein percentages when cheese making is a selection objective.     

Physiological and pathological factors influencing bovine alpha-lactalbumin and beta-lactoglobulin concentrations in milk

Bovine alpha-lactalbumin and beta-lactoglobulin concentrations were determined by radial immunodiffusion in 354 milk samples from uninfected and 98 samples from infected quarters from 42 Holstein-Friesian cows taken at 30, 150, and 270 days of lactation. alpha-Lactalbumin and beta-lactoglobulin concentrations were not affected by quarter location. The alpha-lactalbumin decreased at the end of lactation and in samples collected beyond second lactation. The beta-lactoglobulin concentration increased with stage of lactation. There was a positive correlation between alpha-lactalbumin and beta lactoglobulin (r = .12). Milk from uninfected quarters had mean alpha-lactalbumin and beta-lactoglobulin concentrations of 1.47 and 4.6 mg/ml, respectively. Milk from quarters infected by major pathogens or Corynebacterium bovis had less alpha-lactalbumin. Milk from quarters infected by minor pathogens had less beta-lactoglobulin. There was a negative correlation between alpha-lactalbumin concentration and somatic cell count (r = .31), which was amplified by infection status of quarters. No correlation was noted between somatic cell count and beta-lactoglobulin concentration when considered over the whole sampling period, but the correlation became negative in quarters infected by major pathogens.     

Effects of milk protein variants on the protein composition of bovine milk

The effects of β-lactoglobulin (β-LG), β-casein (β-CN), and κ-CN variants and β-κ-CN haplotypes on the relative concentrations of the major milk proteins α-lactalbumin (α-LA), β-LG, α_S1-CN, α_S2-CN, β-CN, and κ-CN and milk production traits were estimated in the milk of 1,912 Dutch Holstein-Friesian cows. We show that in the Dutch Holstein-Friesian population, the allele frequencies have changed in the past 16 years. In addition, genetic variants and casein haplotypes have a major impact on the protein composition of milk and explain a considerable part of the genetic variation in milk protein composition. The β-LG genotype was associated with the relative concentrations of β-LG (A » B) and of α-LA, α_S1-CN, α_S2-CN, β-CN, and κ-CN (B > A) but not with any milk production trait. The β-CN genotype was associated with the relative concentrations of β-CN and α_S2-CN (A² > A¹) and of α_S1-CN and κ-CN (A¹ > A²) and with protein yield (A² > A¹). The κ-CN genotype was associated with the relative concentrations of κ-CN (B > E > A), α_S2-CN (B > A), α-LA, and α_S1-CN (A > B) and with protein percentage (B > A). Comparing the effects of casein haplotypes with the effects of single casein variants can provide better insight into what really underlies the effect of a variant on protein composition. We conclude that selection for both the β-LG genotype B and the β-κ-CN haplotype A²B will result in cows that produce milk that is more suitable for cheese production.     

Effects of administration of bovine somatotropin on milk yield and composition

Thirty-two Holstein cows were assigned to four treatment groups in a continuous lactation trial to evaluate effects of daily subcutaneous injections of 0 (A), 5.15 (B), 10.3 (C), and 20.6 mg (D) of recombinantly derived bovine somatotropin monomer. Responses were milk yield and composition, component yields, net energy intake, body weight, and somatic cell counts. Treatments began in fall, 28 to 35 d postpartum, and continued for 266 d. Base ration was corn silage and concentrate. Least squares means of milk and constituent yields and gross efficiency of milk production differed among treatments, but percentage fat and protein, energy intake, body weights, and somatic cell counts did not. Least squares means of 3.5% FCM were 21.3 (A), 25.0 (B), 26.2 (C), and 28.5 kg/d (D); fat and protein yields showed similar response to treatment. Milk yields adjusted for pretreatment yields were 21.1 (A), 25.5 (B), 26.5 (C), and 29.3 kg/d (D). Orthogonal contrasts showed lower milk and constituent yields and net energy intake for control (A) than for hormone-injected cows (B, C, D), but no differences were detected between treatments B, C, and D. Gross efficiency of production (3.5% FCM/feed net energy intake) was greater for hormone-injected cows.     

Short communication: Effect of stearoyl-coenzyme A desaturase polymorphism on fatty acid composition of milk

The effect of the stearoyl-CoA desaturase (SCD) gene on milk fatty acid composition was tested. Cows of 3 breeds of northern Italy, Piedmontese, Valdostana, and Jersey, were genotyped at exon 5 of the SCD gene. This has been suggested as a primary candidate gene to change the proportion of saturated vs. unsaturated fatty acids in milk, wherein a single nucleotide polymorphism (C/T) gives rise to a different AA codon. It was possible to ascribe a reduced desaturase activity to the T allele only in the case of caproleic and myristoleic fatty acids. In contrast with the findings of SCD effects on carcass fat, it was not possible to confirm the higher desaturation activity of this single nucleotide polymorphism on long-chain fatty acids, due to the different pathways that originate milk fatty acids of different carbon length; long-chain fatty acids are highly influenced by the complex metabolic events that affect the ingested nutrients during their transfer to milk fat.     

Effects of pasture allowance on the yield and composition of milk from cows of different beta-lactoglobulin phenotypes

The objective of this study was to determine whether the differences in the composition of milk from cows of different beta-lactoglobulin beta-LG) phenotypes are affected by the amount of pasture available and, hence, pasture dry matter intake. Twenty-two Friesian cows of each of the AA and BB variants of the beta-LG phenotype were subjected to ad libitum grazing or restricted grazing in crossover experiments during spring (early lactation, approximately 60 d in milk) and summer (mid to late lactation, approximately 180 d in milk). Milk samples were collected from each cow at the end of each 8-d treatment period and analyzed for composition. Cows of the AA variant of the beta-LG phenotype had higher concentrations of whey protein and beta-LG, but lower concentrations of casein (CN), alpha-CN, kappa-CN (summer only), and BSA, than cows of the BB variant. Compared with cows with a restricted allowance, cows grazing ad libitum had higher milk yields and concentrations of protein, casein, whey protein, and all individual proteins except BSA and immunoglobulin. There were no interactions between effects of pasture allowance and phenotype on milk yield or composition. The data show that having adequate pasture for grazing cows is important not only to maximize milk yield, but also to optimize concentrations of protein and casein, and hence the manufacturing potential of milk. Further, the differences in composition of milk from cows of differing beta-LG phenotypes persisted during short-term restrictions in pasture allowance, and between spring and summer.     

High pressure-induced denaturation of alpha-lactalbumin and beta-lactoglobulin in bovine milk and whey: a possible mechanism

In this study, high pressure (HP)-induced denaturation of alpha-lactalbumin (alpha-la) and beta-lactoglobulin (beta-lg) in dairy systems was examined. In both milk and whey, beta-lg was less baroresistant than alpha-la; both proteins were considerably more resistant to HP-induced denaturation in whey than in milk. HP-induced denaturation of alpha-la and beta-lg increased with increasing proportion of milk in mixtures of milk and whey. Addition of a sulphydryl-oxidising agent, KlO3, to milk or whey increased HP-induced denaturation of beta-lg, but reduced the denaturation of alpha-la. Denaturation of both alpha-la and beta-lg was prevented by adding a sulphydryl-blocking agent, N-ethylmaleimide, to milk or whey prior to HP treatment, highlighting the crucial role of sulphydryl-disulphide interchange reactions in HP-induced denaturation of alpha-la and beta-lg. Removal of colloidal calcium phosphate from milk also reduced HP-induced denaturation of alpha-la and beta-lg significantly. The higher level of HP-induced denaturation of alpha-la and beta-lg in milk than in whey may be the result of the abscence of the casein micelles and colloidal calcium phosphate from whey, which facilitate HP-induced denaturation of alpha-la and beta-lg in milk.     

Effect of beta-lactoglobulin and precipitation of calcium phosphate on the thermal coagulation of milk

The effect of beta-lactoglobulin and heat-induced precipitation of calcium phosphate on the pH dependence and mechanism of thermal coagulation of milk throughout the pH range 6.3-7.3 was studied using serum protein-free milk and sodium caseinate as models for micellar and non-micellar milk protein systems respectively. It appears that the specific effect of beta-lactoglobulin at the pH of maximum stability may be related to its ability to chelate calcium. The effect of beta-lactoglobulin at the pH of minimum stability does not appear to be directly related to heat-induced dissociation of K-casein or micellar integrity but may be due to its ability to sensitize casein micelles to heat-induced precipitation of calcium phosphate, by increasing micellar hydrophobicity. The extent of heat-induced precipitation of calcium phosphate, as a function of pH, is an inverse reflection of the pH dependence of heat stability. Micellar integrity appears to play a critical role in the heat stability of milk but for reasons not previously appreciated.     

Genetic variants of beta-lactoglobulin gene and its association with milk composition traits in riverine buffalo

A study was carried out to determine genetic variants of beta-lactoglobulin gene and to explore associations between these and milk composition traits in riverine buffalo. Single strand conformation polymorphism was employed to detect the genetic variants of the gene. Two fragments of this gene i.e. 119 bp of exon I and 400 bp spanning exon IV and intron IV were included in the study. For 119 bp fragment, three alleles namely, A, B and C were observed in all the buffalo breeds whereas four alleles (A, B, C and D) were detected for 400 bp fragment. The frequency distribution of alleles was different in different breeds of buffaloes for both the fragments. For exon I fragment, the milk composition traits such as total SNF, protein, solid, fat and whey protein yield were found to be significantly (P<0.05) associated with genotypes in Murrah and Bhadawari buffalo whereas in Mehsana breed genotypes were significantly (P<0.05) co-related with total SNF, solid and fat yield. Genotypes of 400 bp fragment, only total fat yield in Mehsana buffalo was found to be significantly (P<0.05) associated with genotypes.     

Effect of recombinant bovine somatotropin on milk production and composition of cows with Streptococcus uberis mastitis.

The protective effect of bovine somatotropin (bST) during experimental Streptococcus uberis mastitis in cows was studied. The left quarters of 10 cows were infected with 500 cfu of S. uberis O140J. Five cows were subcutaneously treated with 500 mg of recombinant bST 7 d before and after infection, and 5 control cows received the excipient. In the treated cows, total milk production significantly increased after the first and second bST treatments. After infection, milk production decreased 24 and 40% in the infected quarters, 6 and 14% in the uninfected quarters, and 15 and 28% overall for treated and control cows, respectively. In the bST group, milk production was completely restored after 3 wk, but, in the control group, total production and the production of the infected quarters remained lower than preinfection production. The increase in somatic cell count occurred earlier and more rapidly in the control group, and the return to normal values was also more rapid in these cows. The amount of bacteria in milk was higher in the control cows. Changes in milk composition, such as lactose, protein, fat, Na+, K+, and Cl-, were significantly more pronounced in the control cows. Also, clinical symptoms were more prominent in the control cows. Somatotropin protected the mammary gland from excessive production losses and compositional changes during a subsequent episode of experimentally induced Streptococcus uberis mastitis and significantly improved the normalization of production and composition, which indicates a beneficial effect on the restoration of the integrity of the blood-milk barrier.     

Effect of modified dry period lengths and bovine somatotropin on yield and composition of milk from dairy cows

Dry periods of 40 to 60 d have been an industry standard because dry periods <40 d have resulted in reduced milk yields in the subsequent lactation by 10 to 30%. However, recent research has demonstrated no production losses for cows given a 30-d dry period. The current study evaluated milk production effects of shortened or omitted dry periods for cows at mature-equivalent production >12,000 kg of milk and treated with bovine somatotropin (bST). The study used 2 commercial dairies and one university dairy and included 4 treatments. Five multiparous and 5 primiparous cows from each farm were assigned to each treatment: 1) 60-d dry period, label use of bST (60DD); 2) 30-d dry period, label use of bST (30DD); 3) continuous milking, label use of bST (CMLST); and 4) continuous milking with continuous use of bST (CMCST). Per label, bST use started at 57 to 70 d in milk and ended 14 d before drying (60DD and 30DD) or expected calving date (CMLST). In primiparous cows, average milk yields during the first 17 wk of lactation were reduced for cows on treatments 30DD, CMLST, and CMCST vs. the 60DD treatment. (38.3, 35.1, and 37.5 vs. 44.1 +/- 1.3 kg/d, respectively). For multiparous cows, respective milk yields did not differ (46.6, 43.4, 46.5, and 47.7 +/- 2.1 kg/d). Shortened or omitted dry periods may impede mammary growth in primiparous cows, resulting in reduced milk yield in the subsequent lactation. In contrast, a shortened or omitted dry period with either bST protocol did not alter production in multiparous cows treated with bST. Quality aspects of prepartum milk and colostrum require additional characterization. For multiparous cows, milk income generated for short dry periods or for continuous milking might increase their profitability. At 17 wk of the subsequent lactation, estimates of the cumulative net margins of multiparous cows on the 30DD treatment and continuous milking treatments exceeded those of cows on the 60DD treatment by 40 dollars to 60 dollars per cow.     

Relationship between composition and stability of bovine milk lysozyme.

The amino acid analysis, peptide mapping, and heat stability of bovine milk lysozyme are presented. The bovine milk lysozyme molecule contains approximately 154 amino acids and is strikingly different in amino acid content from human milk lysozyme and egg white lysozyme. Tryptic hydrolysis yielded 26 peptides, all of which are unique from tryptic peptides of human milk lysozyme and egg white lysozyme. In addition, bovine milk lysozyme was more heat stable than human milk lysozyme at pH 4.0 but more labile at pH 7.0 and 9.0. Possible explanations for the differences in heat stability are discussed.     

Effect of a prolonged-release formation of N-methionyl bovine somatotropin (sometribove) on milk composition.

Forty Holstein cows received bi-weekly injections of prolonged-release formulation of bST, and 39 received bi-weekly injections of excipient, in a study to evaluate the effects of long-term bST administration on milk composition and component production. Injections began at 60 +/- 3 d postpartum. Administration of bST increased production of milk and of all measured components. Concentrations of lactose (4.85 and 4.81%), fat (3.76 and 3.67%), total solids (12.57 and 12.44%), SNF (8.83 and 8.75%), casein (2.56 and 2.53%), and true protein (3.13 and 3.08%) were similar in milks from cows receiving bST and excipient, respectively. Percentages of NPN (times 6.38) and total protein were greater in milk from bST-treated cows (.179% NPN and 3.32% total protein) compared with milk from cows injected with excipient (.172% NPN and 3.24% total protein). Use of bST did not change the relative percentages of alpha s-casein, beta-casein, kappa-casein, beta-lactoglobulin, alpha-lactalbumin, or casein proteolysis products. A cyclical pattern of milk production, component production, and composition within each 14-d injection interval was observed. This suggests that a diminishing amount of bST was delivered during the latter third of each injection interval. There were no effects of bST on milk composition that would be of any practical significance to dairy product manufacturers or consumers.     

Effect of preservatives on milk composition determination

The main objective of our study was to determine the effect of different concentration of four different preservatives, potassium dichromate, Azidiol, Bronopol and Microtabs II, on the results of laboratory analyses of milk using mid-infrared spectrometry. The final concentrations of the preservatives in the raw cows' milk samples were 0.005%, 0.01%, 0.05%, 0.1%, 0.5% and 1% respectively. We analysed unpreserved, fresh bulk raw cows' milk 2 and 24 h after milking. The effect of preservatives on the composition of the milk was studied 24 h after preservation. The experiment was replicated 10 times. Each experiment was carried out after the calibration of the mid-infrared spectrophotometer using calibration samples preserved with 0.02% Bronopol. We found that the concentration of the preserving agent had a significant effect (P < 0.005) on the results of the laboratory analyses; therefore, the proper concentration of preservative has to be used for routine sample preservation.     

Effect of kappa-casein and beta-lactoglobulin loci on milk production traits and reproductive performance of Holstein cows

The effects of kappa-casein (kappa-CN) and beta-lactoglobulin (beta-LG) loci on milk production traits (milk, fat, protein, and lactose yield, fat, protein, and lactose content) and reproductive performance (gestation length, calving interval, age at first and second calving, number of services per conception) was estimated for 278 Holstein cows in the first 2 lactations. Genotypes of kappa-CN and beta-LG were determined by alkaline and acidic polyacrylamide gel electrophoresis. Milk production was recorded daily. Single-trait, mixed, linear models were used for the statistical analysis of the data. Results indicated that kappa-CN genotypes affected significantly protein yield and content (genotype AB > genotype AA). A tendency for increased milk and fat yield of animals having AB kappa-CN genotype was also found. Fat content and lactose yield and content were not affected. In the beta-LG system, significant differences were detected for milk yield (AB > AA), fat yield (BB and AB > AA), fat content (BB > AA and AB), and lactose yield (AB > AA). A tendency for higher protein yield was also observed (AB > AA). The beta-LG locus had no significant effect on protein and lactose content. No associations between polymorphisms at the kappa-CN locus and reproductive performance were found. There was a tendency, however, for cows with AB genotype to have older age at first and second calving. In the beta-LG system, cows with AA genotype had significantly shorter gestation length than did those with AB or BB genotype. No differences were detected between beta-LG polymorphisms for the other reproductive traits.