Effect of pH on the gelation properties of skim milk gels made from plant coagulants and chymosin

The effect of three milk pH values, 6.0, 6.3 and 6.7, on gelation properties was monitored by small amplitude oscillatory rheology as well as a large deformation (yield) test for gels induced by the plant coagulants, Cynara cardunculus L. and Cynara humilis L., and chymosin. Gel microstructure was studied using confocal scanning laser microscopy. For each coagulant, a decrease in pH of milk resulted in a decrease in gelation time (tg), and an increase in the rate of increase in storage modulus (G'). At pH 6.0 and 6.3, plant coagulant-induced gels reached a maximum value in G' (G'max) followed by a decrease in G' values during the rest of the experiment, reflecting higher proteolytic activity of plant coagulants towards caseins as determined by SDS-PAGE. Gels produced at pH 6.0 and 6.3, exhibited a minimum in loss tangent (tan delta) followed by slight increase in tan delta during gel aging, that may have been associated with faster rearrangements of the gel network structure. In gels aged for approximately 6 h, the values for G', tan delta at low frequency (0.006 Hz) and yield stress were higher for chymosin than for plant-induced gels. For gels with the same pH value, no major differences were observed in microstructure between coagulants. Gels made at low pH values (6.3 and 6.0) appeared to have a denser or more interconnected structure than gels made at pH 6.7. Our results suggest that, at a low pH, the type of coagulant used in gelation is likely to have a considerably impact on gel/cheese structure.     

Influence of denaturation and aggregation of β-lactoglobulin on rennet coagulation properties of skim milk and ultrafiltered milk

Skim milk and ultrafiltered milk were heated at temperatures in the range 80–140 °C for 4s in a UHT plate heat exchanger. The extent of denaturation of β-lactoglobulin and its association with the casein micelles increased with increase in heating temperature; the extent of association was markedly less than the amount that denatured. There was no noticeable difference in the extent of denaturation and association of β-lactoglobulin between skim milk and the corresponding ultrafiltered milk. The coagulation properties of renneted heated milks were measured by dynamic oscillation rheometry. Gelation times and storage modulus (G′) could be related to the extent of denaturation of β-lactoglobulin and its association with the casein micelles. Denaturation of β-lactoglobulin up to 60% had no effect on the gelation time, but additional denaturation markedly increased the gelation times. In contrast, G' decreased gradually with increase in the extent of denaturation of β-lactoglobulin and its association with casein micelles. The effects of β-lactoglobulin denaturation and association on coagulation properties were less pronounced in ultrafiltered milk. G' values for ultrafiltered milk were markedly higher than the corresponding skim milks under all heating conditions.     

Growth of lactic acid bacteria in highly concentrated ultrafiltered skim milk retentates

Buffer capacity of ultrafiltered skim milk retentates at various protein concentrations and growth of direct set, frozen concentrated lactic starter cultures in such retentates were studied. Maximum buffering occurred at approximately pH 5.1 to 5.3. An average .48% lactic acid concentration was required to reduce the pH of plain skim milk to 4.6 compared with 1.01% for skim milk retentates concentrated 2.3:1 and 1.14% for skim milk retentate concentrated 2.6:1. Skim milk retentates concentrated 4.3:1 and 5.8:1 were unable to attain pH 4.6 even when titratable acid was greater than 1.8%. Lactic acid required to reduce pH to 4.6 for the two lower concentrated retentates (2.3:1 and 2.6:1) were 1.85 and 2.45%. Time to attain pH 4.6 was a function of the bacterial cell concentration of the cultures and the total protein level of retentates. Starter organism growth was unaffected by high total solids or ash of retentates. Growth rate and lactose metabolism decreased markedly below pH 5.2 at which point bacterial population was 10(9) cfu/ml.     

Effects of hydrodynamic cavitation and temperature on nanofiltration performance for concentrating ultrafiltered skim milk

The performance of nanofiltration (NF) as influenced by hydrodynamic cavitation (HC) and filtration temperature during the concentration of milk protein concentrate (MPC) was investigated. Pasteurised skim milk was concentrated using ultrafiltration (UF) to prepare UF retentate (MPC80, 20% total solids, TS), which was then further concentrated using NF at 22°C and 50°C with or without HC treatments until permeate flux declined to 0.1 L/m²/h. Results showed that UF retentate can be concentrated to higher TS (up to 31.5%) at higher filtration temperature or by applying HC, with synergistic effect in combination of both treatments, during NF.     

Antagonism between Listeria monocytogenes and lactococci during fermentation of products from ultrafiltered skim milk.

Tyndallized samples of unfiltered skim milk and retentate (concentrated fivefold or twofold by volume) and permeate from UF skim milk were inoculated with 5.5 x 10(3) to 1.5 x 10(5) cfu/ml of Listeria monocytogenes strains California or V7 together with 4 x 10(7) to 2.3 x 10(8) cfu/ml of mesophilic lactic acid bacteria. Numbers of L. monocytogenes (McBride Listeria agar) and lactic acid bacteria (all purpose Tween agar) were determined after 0, 6, 12, 24, 30, and 36 h of incubation at 30 degrees C. Lactic acid bacteria significantly inhibited or inactivated L. monocytogenes in all three products. Inactivation was greater in permeate (6.77 orders of magnitude) than in unfiltered skim milk (3.67 orders of magnitude) or in retentate (4.21 orders of magnitude). Degree of inactivation in retentate was related to the extent of concentration. Inactivation was not complete, and L. monocytogenes survived in these products during fermentation for up to 36 h. When fermented products were refrigerated (4 degrees C), L. monocytogenes survived for 4 to 6 wk in skim milk, 3 to 5 wk in retentate, and 1 wk in permeate. At refrigeration temperature, length of survival was dependent on type of product and strain of the pathogen.     

Characterisation of potential milk coagulants from Calotropis gigantea plant parts and their hydrolytic pattern of bovine casein

The steady increase in world cheese demand keeps the search for appropriate coagulating enzymes as rennet substitutes in the scientific focus. This work reports the milk-clotting activities of aqueous crude enzyme (CE) from different parts of Calotropis gigantea using skim milk as substrate. CE from latex exhibited high caseinolytic (86.445 ± 1.055 U/ml) as well as milk-clotting activity (450 U/ml) when compared to other parts. Significant milk clotting index (MCI) was presented by crude enzyme of latex followed by stem, flower and leaf in the decreasing order (p < 0.05). CE from all parts showed an optimum activity at 37 °C, pH 5.5 and 10 mM calcium chloride concentration with excellent pH (4.5–6.5) and thermal stability (30–60 °C). Hydrolysis pattern of casein components by CE during 1 h incubation, as assessed by Tricine–SDS-PAGE, and subsequent peak analysis revealed CE from stem to be closely similar to that of rennin. However, extensive casein hydrolysis was observed when incubated with crude latex enzyme. Casein bands of CE and rennin tended to disappear as incubation progressed to 24 h. Proteolytic degradation was found to be advanced and resulted in complete breakdown of casein fractions unravelling the importance of incubation time as a possible essential requisite for controlled and appropriate casein hydrolysis. The study provides evidence of rennin-like activity associated with C. gigantea plant parts which may serve as a promising source of vegetable milk coagulant.     

Effect of bovine skim milk and whey on monocyte function

Previous studies have documented the ability of bovine milk to inhibit lymphocyte proliferation in response to mitogens. It is not known whether inhibition of lymphocyte proliferation is mediated through the action of monocytes. To address this question, we examined the ability of bovine skim milk and whey to affect monocyte function with emphasis on expression of major histocompatibility class II antigens and production of interleukin-1 by monocytes. Data showed that expression of major histocompatibility complex class II molecules and production of interleukin-1 by monocytes were not altered when monocytes were cultured in the presence of bovine skim milk or whey. Thus, it is unlikely that the suppressive effect of milk on lymphocyte proliferation could be mediated through alterations in the expression of major histocompatibility class II molecules or in production of interleukin-1 by monocytes. The role of other monocyte antigens or secretory products, however, should also be evaluated.     

Relationships between radioimmunoassays of alpha lactalbumin and prolactin in bovine skim milk

A radioimmunoassay developed for alpha-lactalbumin was sensitive between 5 and 140 ng alpha lactalbumin. Addition of increasing volumes of milk to assay tubes progressively decreased binding of iodine-125-labeled alpha-lactalbumin to the antisera in a manner which paralleled the binding curve generated by increasing concentrations of standard alpha-lactalbumin. The addition of 10, 20, 30, or 40 ng of alpha-lactalbumin to diluted milk samples gave 90, 100, 105, and 102% recoveries. Alpha-lactalbumin antisera did not crossreact with 1000 ng of bovine casein, blood serum albumin, beta-lactoglobulin, prolactin, or growth hormone. Milk from each of approximately 100 Holstein Friesian cows at different stages of lactation was sampled monthly for 12 mo. Concentrations of alpha-lactalbumin (1.63 mg/ml) and prolactin (24.9 ng/ml) in samples of skim milk collected in the 1st mo of lactation were greater than those in the remaining months of lactation. Monthly concentrations of alpha-lactalbumin and prolactin in skim milk did not change significantly during seasons of the year. The correlation between concentrations of prolactin and alpha-lactalbumin pooled within subclasses of month of lactation within month of year was .08 for 1125 pairs.     

Effect of mastitis on proteolytic activity in bovine milk

Proteolytic activity of milk was studied before, during, and after experimental-induced mastitis. An inoculum of Streptococcus agalactiae was infused into one quarter of each udder of six cows to elicit an infection. Bacteriological cultures and SCC of milk were used to monitor infection status. Sodium dodecyl sulfate-PAGE was used to measure proteolytic activity of milk. Inhibitor 6-amino-n-hexanoic acid was used to determine the relative proportion of plasmin and nonplasmin proteolytic activity of milk. Somatic cell count, total milk proteolytic activity, and nonplasmin proteolytic activity were higher in infected quarters than in quarters preinfection. After elimination of infections, SCC and nonplasmin proteolytic activity decreased to preinfection amounts. Total proteolytic activity of milk decreased after infections were cured but remained significantly higher than preinfection activity. This postinfection proteolytic activity in milk may be due to an increase in milk plasmin activity. Our data suggest that detrimental effects of mastitis on milk quality can continue after infection has been eliminated and milk SCC have returned to low values.     

Changes in the calcium cluster distribution of ultrafiltered and diafiltered fresh skim milk as observed by Small Angle Neutron Scattering

The effect of ultrafiltration and diafiltration on the distribution of the calcium phosphate clusters of the casein micelle was investigated using Small Angle Neutron Scattering (SANS). In the case of ultrafiltration, fresh skim milk was subjected to concentration using membrane filtration up to 5× its original volume, the retentate was rediluted with its corresponding serum and subsequently dialyzed against reconstituted milk powder dispersed in D2O/H2O (UF 5×(D)). In the case of diafiltered samples, the samples were concentrated adding water (diafiltration) at two different levels (DF 2·5× or DF 5×) and then redispersed with D2O/H2O. In the DF 5× case, the serum components were diluted to less than 1% of their original concentration. For analysis, all samples had the same volume fraction of dispersed casein micelles (φ=0·1), which is that of the control, unprocessed skim milk. A peak in the SANS data was observed in fresh skim milk at a scattering vector, qo, of 0·034 ?-1 (directly proportional to the reciprocal characteristic length), in agreement with previous literature results. Neutron data on the ultrafiltered, UF 5×(D) and diafiltered, DF 2·5× and DF 5× milk samples showed a progressive decrease in the intensity of the peak but invariance in qo. These results, combined with the determination of soluble and insoluble calcium in the samples, suggest a progressive and irreversible removal of calcium from within the micelle during membrane filtration of milk. Using SANS it was possible to clearly show changes in the micellar calcium clusters that may not otherwise be fully determined by only measuring the amount of total and insoluble calcium in milk.     

Effect of lactose content on dielectric properties of whole milk and skim milk

Dielectric properties (dielectric constant ε′ and dielectric loss factor ε′′) of whole milk and skim milk with the lactose content of 4.56–6.44% and 4.57–6.47%, respectively, were measured over the frequency range of 20–4500 MHz at 25 °C using a vector network analyzer. The results showed that ε′ decreased with increasing frequency, and ε′′ changed with V shape and its minimum was noted at about 2000 MHz. Whole milk had lower dielectric properties than skim milk at almost same lactose content and a given frequency. ε′ had weak positive linear relationship with lactose content for whole milk, but had negative linear relationship for skim milk. No matter for which milk, ε′′ had very good negative linear relationship with lactose content below 1000 MHz and had good positive linear relationship above 2300 MHz. The study provides information on developing rapid lactose detector for milk in future.     

Lipolytic activity with the membrane fraction of bovine skim milk.

Colloidal phosphate-free skim milk was subjected to gel filtration on Sepharose 4B. Lipolytic activity was observed in the membrane material eluted in the void volume fraction and in the protein fraction representing a broad range of molecular weights.     

Physico-chemical properties of skim milk retentates from microfiltration.

Physico-chemical properties of retentates obtained from selective concentration of skim milk up to 8 times its original weight using a microfiltration system were studied. The effects of process variables, namely concentration (8.6 to 27 wt.%), temperature (20 to 50 degrees C) and pH (6.0, 6.3, and 6.5) on density (rho), apparent viscosity (mu(a)), consistency coefficient (K), flow behavior index (n), and activation energy (Ea) of the retentates were examined. Depending on pH, retentates showed significant increase in apparent viscosity, deviated from classical Newtonian behavior and exhibited shear-thinning between 11 to 17% solids concentration. No yield stress was detected in the range of concentration studied. The power law parameters (n and K) followed a similar trend. An Arrhenius-type equation described well the effect of temperature on apparent viscosity. Although activation energy increased 120 to 130% for a threefold increase in solids concentration, it was not significantly different from that of other types of concentrated milk at approximately the same concentration. Increasing solids were responsible for change in flow properties with concentration, while the effect of pH was attributed to differential protein (primarily casein) retention and the change in solvation properties and voluminosity of casein micelles. Models relating concentration, temperature, and pH to retentate apparent viscosity and consistency coefficient were identified. Skim milk microfiltration with in-process pH adjustment produces retentates depleted in whey proteins and calcium with significantly altered properties.     

Effects of selected properties of ultrafiltered spray-dried milk powders on some properties of chocolate

The effects of selected properties of spray-dried milk fat powders on chocolate were determined. Milk powders produced from control or ultrafiltered (UF) milks with various levels of fat were blended with skim milk powder to give a standard 26 g fat 100 g⁻¹ powder. Particle size of the chocolate mixes after refining decreased as the fat content and free-fat content of the powders increased. Despite this, increasing fat and free-fat contents of powders reduced the Casson viscosity of the subsequent molten chocolates. Casson viscosities using powders from control or UF milks were similar, but decreased as the particle size of powders increased and particle size after refining the chocolate mix decreased. Casson yield value and hardness decreased as fat content of powders increased. Casson yield value increased with vacuole volume of powders. It is possible to alter important properties of chocolates using milk powders of varying fat contents, free-fat contents and particle sizes.     

Presence of lactoferrin in a bovine skim milk fraction with acid phosphatase activity

The aim of this study was to determine the principal active molecules in the acid phosphatase (AcP) fraction of skim milk origin using immunostaining and AcP staining. The AcP fraction was separated from skim milk at 0.38 m NaCl using carboxymethyl cellulose chromatography at pH 5.2. The molecular mass of the active molecule in AcP fraction was estimated to be 80 kDa by immunostaining and AcP staining. The 80 kDa protein was analyzed by a protein sequencer, using the automated Edman degradation method; the first thirteen N-terminal amino acid sequence obtained were shown to be APRKNVRWXTIXQ. For that amino acid sequence, there was 84% (11/13 residues) homology with the amino acid sequence of bovine lactoferrin (LF). The AcP fraction and commercial LF showed a similar AcP activity profile, having an optimum pH of 4.5 and temperature of 60 °C. Thus, the AcP fraction from bovine skim milk was isolated and the principal active molecule present was tentatively identified as LF.     

Coagulation properties of ultrafiltered milk retentates measured using rheology and diffusing wave spectroscopy

The effect of concentration of milk by ultrafiltration on renneting has been widely studied as it is of great interest in dairy technology. Although a number of reports are available on the texture and microstructure of the milk gels formed at various concentrations, very little is understood on the effect of concentration on the stages preceding aggregation, or how concentration may affect the interactions between micelles. This study aims to investigate the renneting behavior of milk concentrated by ultrafiltration (without diafiltration) to 3× and 5× (v/v) and compare it to that of skim milk. The scattering properties of the casein micelles under quiescent conditions suggest that they deviate from hard-sphere behavior at 5× concentration (micelle volume fraction, ? = 0.5). The release of the caseinomacropeptide during renneting was not significantly different amongst the three different casein concentrations tested (1×, 3×, and 5×). No significant differences were also noted in the rennet coagulation time as detected by both diffusing wave spectroscopy and rheology. Concentrated milk samples formed significantly (p-value < 0.05) stiffer gels than regular milk due to an increased number of bonds in the network. The level of milk concentration also accelerated a change in the spatial distribution and rate of change of turbidity of the micelles because of a decrease in the overall inter-particle distance and increased collision frequencies. This in situ investigation of concentrated milk samples suggested that the changes in rennet coagulation with concentration are merely a cause of crowding effects.     

脱脂乳的不同热处理对其酸奶凝胶流变学性质的影响

研究了复原脱脂乳经不同加热处理后其酸奶凝胶形成过程中流变学特性的变化,复原脱脂乳经７０～９０℃处理１０～２５ｍｉｎ,用Ｄ－葡萄糖－δ－内酯酸化成胶,用流变学方法动态监测凝胶性能。并监测酸化过程和成胶时间,结果显示,不同条件下自理后的脱脂乳在酸化过程中,ｐＨ值并未改变,但产储存模量（Ｇ′）和损失模量（Ｇ″）随受质变程度的增加而增加,只是当加热温度为９０℃时反而随加热时间的延长而降低,在８５℃加热２５     

Factors affecting ability of skim milk to support phagocytosis by bovine polymorphonuclear leukocytes

Milk samples from quarters were collected from 48 Holstein cows at 1.5, 3, 21, and 35 wk postpartum and at 1 wk after drying off. Foremilk samples were obtained for bacteriological examination, somatic cell count of milk, and for assay of phagocytosis. Thirty-two cows in first lactation and 16 in third lactation produced by two criteria of sire selection (high milk versus multiple trait) were studied. In vitro phagocytosis was assayed four times on each original quarter milk sample, yielding 3,840 determinations. Variance components were large for cow-associated variation. Cow, stage X cow, and cow X sample dilution were 18.7, 20.6, and 6.0% of total variance. Phagocytosis was 9% higher in skim milk from third lactations than from first. Phagocytosis was highest in dry period samples, followed by 1.5, 35, 3, and 21-wk postpartum samples. Milk somatic cell count tended to be related more closely to phagocytosis than was current bacteriological status of the quarter. Skim milk from genetically higher producing cows was less conducive to phagocytosis than skim milk from genetically lower producing cows, possibly because of dilution.