Variation in Coagulation Properties of Milk from Individual Cows,

Milk samples from 50 Holstein cows were tested monthly for 10 mo for total protein, casein, fat, somatic cells, and pH. A Formagraph was used to measure chymosin coagulation properties. Significant variations in coagulation time and curd firmness were observed in relation to period of lactation, individual cows, and milk pH. A high negative correlation coefficient (?.86) was observed between coagulation time and curd firmness measured 30 min after addition of chymosin. The mean coagulation time generally increased as lactation progressed and milk yield decreased. Curd firmness was generally greatest in midlactation samples.Milk from 38% of the cows did not coagulate in 30 min 1 mo prior to their dry periods. The frequency of failure to coagulate was 68% in winter and 32% in fall. Milk pH was the most significant factor that affected coagulation time and curd firmness. Simulated cheese making procedures were utilized to estimate recovery of fat and protein in curd. Curd yield calculated from the recovery data ranged from 5.4 to 14% with a mean of 9.2%.     

Estimation of Residual Pepsin and Chymosin Activity in Curd,

Pasteurized milk (225 g) adjusted to pH 6.2 was set with 3.5 milk clotting units of chymosin (EC 3.4.23.4). The same amount of milk at pH 5.8 was set with 3.5 milk clotting units of porcine pepsin (EC 3.4.23.1). Fifteen minutes after clotting, the curd was broken, and curd and whey were separated by centrifugation at 3500 × g for 20 min. The curd (30 g) was extracted at pH 6.8 in 450 ml water or at pH 6.2 (chymosin) or 5.8 (pepsin) in 450 ml 1 M sodium chloride.Chymosin was completely released from the curd and accounted for by both methods of extraction. Pepsin was completely released and accounted for after extraction in 1 M sodium chloride at pH 5.8 but was partly inactivated during extraction at pH 6.8.Assay of curd extracts and whey by a linear agar diffusion test accounted for 102 ± 6% of the pepsin activity added to milk when the curd was extracted in 1 M sodium chloride. Extraction at pH 6.8 allowed recovery of only 63% of the activity. Chymosin recovery was 100 ± 5% by both methods of curd extraction.     

Genetic analysis of coagulation properties,curd firming modeling,milk yield,composition, and acidity in Sarda dairy sheep

Sheep milk is an important source of food, especially in Mediterranean countries, and is used in large part for cheese production. Milk technological traits are important for the sheep dairy industry, but research is lacking into the genetic variation of such traits. Therefore the aim of this study was to estimate the heritability of traditional milk coagulation properties and curd firmness modeled on time t (CF_t) parameters, and their genetic relationships with test-day milk yield, composition (fat, protein, and casein content), and acidity in Sarda dairy sheep. Milk samples from 1,121 Sarda ewes from 23 flocks were analyzed for 5 traditional coagulation properties by lactodynamographic tests conducted for up to 60 min: rennet coagulation time (min), curd-firming time (k₂₀, min), and 3 measures of curd firmness (a₃₀, a₄₅, and a₆₀, mm). The 240 curd firmness observations (1 every 15 s) from each milk sample were recorded, and 4 parameters for each individual sample equation were estimated: rennet coagulation time estimated from the equation (RCT_eq), the asymptotic potential curd firmness (CF_P), the curd firming instant rate constant (k_CF), and the syneresis instant rate constant (k_SR). Two other derived traits were also calculated (CF_max, the maximum curd firmness value; and t_max, the attainment time). Multivariate analyses using Bayesian methodology were performed to estimate the genetic relationships of milk coagulation properties and CF_t with the other traits; statistical inference was based on the marginal posterior distributions of the parameters of concern. The marginal posterior distribution of heritability estimates of milk yield (0.16 ± 0.07) and composition (0.21 ± 0.11 to 0.28 ± 0.10) of Sarda ewes was similar to those often obtained for bovine species. The heritability of rennet coagulation time as a single point trait was also similar to that frequently obtained for cow milk (0.19 ± 0.09), whereas the same trait calculated as an individual equation parameter exhibited larger genetic variation and a higher heritability estimate (0.32 ± 0.11). The other curd firming and syneresis traits, whether as traditional single point observations or as individual equation parameters and derived traits, were characterized by heritability estimates lower than for coagulation time and for the corresponding bovine milk traits (0.06 to 0.14). Phenotypic and additive genetic correlations among the 11 technological traits contribute to describing the interdependencies and meanings of different traits. The additive genetic relationships of these technological traits with the single test-day milk yield and composition were variable and showed milk yield to have unfavorable effects on all measures of curd firmness (a₃₀, a₄₅, a₆₀, CF_P, and CF_max) and t_max, but favorable effects on both instant rate constants (k_CF and k_SR). Milk fat content had a positive effect on curd firmness traits, especially on those obtained from CF_t equations, whereas the negative effects on both coagulation time traits were attributed to the milk protein and casein contents. Finally, in view of the estimated heritabilities and additive genetic correlations, enhancement of technological traits of sheep milk through selective breeding could be feasible in this population.     

Effects of pH, calcium chloride, and chymosin concentration on coagulation properties of abnormal and normal milk

Individual Holstein cow milk samples were selected for good and poor chymosin-coagulation characteristics. The effect of pH adjustment, addition of .02% calcium chloride, and variation in chymosin concentration on coagulation properties of good and poor coagulating samples was evaluated. Pooling 50% good and 50% poor coagulating samples did not improve the average coagulation properties of the poor samples. Reducing milk pH to 6.3 caused a significant decrease in coagulation time but a less marked increase in curd firmness. The greatest increase in curd firmness was obtained by a combination of reducing milk pH, addition of .02% calcium chloride, and reducing chymosin concentration. High-chymosin concentration at reduced pH decreased coagulation time without substantially increasing curd firmness. Curd disintegration was more apparent at high-chymosin concentration in the poor coagulating samples.     

Effects of Somatic Cell Counts and Milk Composition on the Coagulating Properties of Milk

Forty-two Holstein cows were selected to provide monthly milk samples with varying SCC for 1 yr. Coagulating properties of samples measured as rennet clotting time, rate of curd firming, and curd firmness at cutting were determined by a formagraph. Milk samples were analyzed for fat, protein, lactose, total solids, casein, individual caseins, urea, SCC, and pH. Least squares analyses of data, after adjustments were made for the effect of milk composition, indicated that elevated SCC were associated with a significant increase in rennet clotting time and slower rate of curd firming. An increase of SCC from 100,000 to 500,000 SCC/ml resulted in an increase of approximately 2.1 and 2.2% in RCT and K20, respectively. A further increase of SCC to above 1,000,000/ml resulted in an overall increase of 20.7 and 13.84% in RCT and K20, respectively. Regression analyses indicated that K20 was decreased by 5.42 min and curd firmness at cutting was increased by 12.92 mm for every percentage in milk casein. Rennet clotting time, rate of curd firming, and curd firmness at cutting were increased by 3.52, 3.41, min and decreased by 9.45 mm, respectively, for every unit increase in milk pH.     

Breed of cow and herd productivity affect milk composition and modeling of coagulation,curd firming,and syneresis

Milk coagulation properties (MCP) have been widely investigated in the past using milk collected from different cattle breeds and herds. However, to our knowledge, no previous studies have assessed MCP in individual milk samples from several multi-breed herds characterized by either high or low milk productivity, thereby allowing the effects of herd and cow breed to be evaluated independently. Multi-breed herds (n = 41) were classified into 2 categories based on milk productivity (high vs. low), defined according to the average milk net energy yielded daily by lactating cows. Milk samples were taken from 1,508 cows of 6 different breeds: 3 specialized dairy (Holstein-Friesian, Brown Swiss, Jersey) and 3 dual-purpose (Simmental, Rendena, Alpine Grey) breeds, and analyzed in duplicate (3,016 tests) using 2 lactodynamographs to obtain 240 curd firming (CF) measurements over 60 min (1 every 15 s) for each duplicate. The 5 traditional single-point MCP (RCT, k₂₀, a₃₀, a₄₅, and a₆₀) were yielded directly by the instrument from the available CF measures. All 240 CF measures of each replicate were also used to estimate 4 individual equation parameters: RCT estimated according to curd firm change over time modeling (RCT_eq), asymptotic potential curd firmness (CF_P), curd firming instant rate constant (k_CF), and syneresis instant rate constant (k_SR) and 2 derived traits: maximum curd firmness achieved within 45 min (CF_max) and time at achievement of CF_max (t_max) by curvilinear regression using a nonlinear procedure. Results showed that the effect of herd-date on traditional and modeled MCP was modest, ranging from 6.1% of total variance for k₂₀ to 10.7% for RCT, whereas individual animal variance was the highest, ranging from 32.0% for t_max to 82.5% for RCT_eq. The repeatability of MCP was high (>80%) for all traits except those associated with the last part of the lactodynamographic curve (i.e., a₆₀, k_SR, k_CF, and t_max: 57 to 71%). Reproducibility, taking into account the effect of instrument, was equal to or slightly lower than repeatability. Milk samples collected in farms characterized by high productivity exhibited delayed coagulation (RCT_eq: 18.6 vs. 16.3 min) but greater potential curd firmness (CF_P: 76.8 vs. 71.9 mm) compared with milk samples collected from low-productivity herds. Parity and days in milk influenced almost all MCP. Large differences in all MCP traits were observed among breeds, both between specialized and dual-purpose breeds and within these 2 groups of breeds, even after adjusting for milk quality and yield. Milk quality and MCP of samples from Jersey cows, and coagulation time of samples from Rendena cows were better than in milk from Holstein-Friesian cows, and intermediate results were found with the other breeds of Alpine origin. The results of this study, taking into account the intrinsic limitation of this technique, show that the effects of breed on traditional and modeled MCP are much greater than the effects of herd productivity class, parity, and DIM. Moreover, the variance in individual animals is much greater than the variance in individual herds within herd productivity class. It seems that improvement in MCP depends more on genetics (e.g., breed, selection) than on environmental and management factors.     

Effect of genetic polymorphism of milk proteins on rheology of chymosin-induced milk gels

Individual milk samples from 121 cows in mid lactation of the Swedish Red and Swedish Holstein breeds with known protein genotypes of β- and κ-casein and β-lactoglobulin were analysed. Chromatographically pure chymosin was added to skim milk and rheological properties of the gels were measured using a Bohlin VOR Rheometer. Coagulation time (CT) and curd firmness after 25 min (G′₂₅) were registered for each sample. The B allele of κ-casein was associated with improved coagulating properties of milk, whereas the E allele showed a negative effect on these traits. The β-casein A²A² genotype was associated with inferior milk coagulation characteristics. Total protein concentration of milk was positively associated with curd firmness, but showed no association with milk coagulation time.     

A Naturally Buffered Bulk Retentate Starter from Ultrafiltered Milk

Whole milk was ultrafiltered to approximately 4:1 protein concentration, heated to 85°C for 30 min, and cooled to 22°C. It was inoculated with a commercial frozen concentrated lactic starter to give approximately 10⁷ cfu/ml and incubated at 22°C for 12 h. A commercial phage inhibitory medium and 11% nonfat dry milk were used as controls. After 12 h, retentate had significantly higher colony forming units per milliliter (3.2 × 10⁹) and pH (5.21) than phage inhibitory medium (2.5 × 10⁹ and pH 5.02) and nonfat dry milk (2.4 × 10⁹ and pH 4.58). Retentate starter and phage inhibitory medium starter had equal activity in skim milk (.3% developed acidity in 4 h at 32°C) whereas nonfat dry milk starter had significantly lower activity (.26% developed acidity). After a further 8 h incubation at 22°C, retentate starter had the highest pH (4.95) compared with phage inhibitory medium (4.76) and nonfat dry milk (4.51). At this time retentate starter activity was higher (.3%) than phage inhibitory medium (.27%) and nonfat dry milk (.19%). In highly concentrated retentates (3.5:1 and 5:1), retentate starter lowered pH considerably quicker than nonfat dry milk starter.     

Effects of Milk Composition and Genetic Polymorphism on Coagulation Properties of Milk

Milk samples from 31 Holstein cows of different phenotypes for β-casein, κ-casein, and β-lactoglobulin were collected monthly over the entire lactation. These samples were analyzed for total solids, fat, protein, casein fractions, lactose, urea, citric acid, somatic cell count, and pH. Rennet clotting time, rate of firming, and curd firmness as measured by a Formagraph were not significantly influenced by phenotypes for β-casein and κ-casein. Phenotype AA for β-lactoglobulin gave the best clotting time (3.91 min) and firmness of curd (36.30 mm) when compared with AB and BB phenotypes. Relative percentages of the different caseins and α-lactalbumin affected significantly rate of firming and curd firmness at cutting. Amount of κ-casein in milk was the most significant factor that affected curd firmness with a coefficient of regression of 15.96.     

Cheese made from instant infusion pasteurized milk: Rennet coagulation,cheese composition,texture and ripening

Milk subjected to instant infusion pasteurization (IIP) at 72 °C, 100 °C and 120 °C (holding time 0.2 s) exhibited increased rennet coagulation time and decreased curd firming rate for increasing heat treatment temperature, when compared with raw or high temperature short time pasteurized (HTST) milk. However, addition of 4.5 mm or 9.0 mm of calcium restored the impaired rennet coagulation ability. Open texture cheeses produced from IIP milk (100 °C and 120 °C) contained significantly more moisture, had lower pH and shorter texture than similar cheese from IIP at 72 °C and HTST pasteurized milk. Cheese ripening was also affected by heat treatment, and different patterns of casein breakdown and peptide formation resulted from cheeses made from milk treated to IIP at 100 °C and 120 °C compared with cheeses made using IIP at 72 °C or HTST.     

Effect of genetic polymorphism of milk proteins on rheology of acid-induced milk gels

Individual milk samples from 80 cows in mid-lactation of the Swedish Red and Swedish Holstein breeds with known protein genotypes of β- and κ-casein and β-lactoglobulin were analysed for acid coagulation properties. Glucono-δ-lactone (1.5%) was added to defatted, heated (90–95 °C) samples and rheological properties of the gels were measured using a Bohlin VOR Rheometer. Coagulation time (CT) and curd firmness after 4, 8 and 10 h (G₄′, G₈′, and G₁₀′ were registered for each sample. Milk protein composition was analysed by reversed phase HPLC. Concentration of β-lactoglobulin in milk was found to be an important factor for the variation in CT and G′. The A allele of β-lactoglobulin was associated with higher concentrations of β-lactoglobulin in milk compared with B. When no adjustment for β-lactoglobulin concentration was made, there was a significant overall effect of β-lactoglobulin genotype on acid coagulation, where the AA and AB genotypes were associated with better curd firmness compared with BB, whereas at equal β-lactoglobulin concentrations a tendency in the opposite direction was found with a significant and positive effect of BB compared with AB. Lactose concentration of milk had a positive effect on acid coagulation and was shown to improve G′ in milk with low β-lactoglobulin concentrations.     

Coagulation Properties of Abnormal and Normal Milk from Individual Cow Quarters

Individual quarter milk samples from two Jersey and two Holstein herds were tested for chymosin coagulation time and curd firmness in the Formagraph instrument. Normal and abnormal milk samples were identified from somatic cell counts. Conductivity measurements and pH were determined.Coagulation properties of normal and abnormal milk drawn from individual quarters were significantly different. Abnormal milk had longer coagulation times and formed weaker curd than normal milk. Both normal and abnormal Jersey milk had shorter coagulation times and formed firmer curd than normal and abnormal Holstein milk. Coagulation properties of milk samples containing primary or secondary pathogens were not significantly different from each other but were inferior to samples that contained no pathogens.     

Effect of milk fat concentration and gel firmness on syneresis during curd stirring in cheese-making

An experiment was undertaken to investigate the effect of milk fat level (0%, 2.5% and 5.0% w/w) and gel firmness level at cutting (5, 35 and 65 Pa) on indices of syneresis, while curd was undergoing stirring. The curd moisture content, yield of whey, fat in whey and casein fines in whey were measured at fixed intervals between 5 and 75 min after cutting the gel. The casein level in milk and clotting conditions was kept constant in all trials. The trials were carried out using recombined whole milk in an 11 L cheese vat. The fat level in milk had a large negative effect on the yield of whey. A clear effect of gel firmness on casein fines was observed. The best overall prediction, in terms of coefficient of determination, was for curd moisture content using milk fat concentration, time after gel cutting and set-to-cut time (R² = 0.95).     

Effect of Ultrasonic Treatment on the Extraction of Chymosin

Improvement of chymosin (rennin) production is limited by a prolonged extraction process. An ultrasonic method of chymosin extraction was developed to increase efficiency. The application of ultrasonic energy significantly reduced time required for the extraction process as a result of abomasum tissue dispersing, destruction of the cells, intensive blending, separation of particles, and increase of the interphase abomasum-extraction medium. The ultrasonic extraction process significantly increased yield of chymosin.Optimal technological parameters for extraction and properties of extraction medium were established: time of ultrasonic treatment (45 min), intensity of ultrasound (3.34 W/cm²), temperature of extraction medium (15°C), NaCl concentration in solution (5%), ratio of abomasum to extracting solution (1:18), NaCl concentration in solution (5%), ratio of abomasum to extracting solution (1:18), pH of solution (4.2 to 4.3), time of chymosin extract activation (7 h), degree of abomasum shredding (strips 5 to 8 mm wide), and time of abomasum swelling (20 min).     

Induction of Lactation by Two Techniques: Success Rate,Milk Composition,Estrogen and Progesterone in Serum and Milk,and Ovarian Effects

Induction of lactation was attempted in 12 heifers and 12 cows with estradiol benzoate (.011 mg/kg body weight per day) subcutaneous for 10 days or that plus progesterone (.1 mg + .25 mg/kg body weight per day) for 7 days. Milking commenced on day 20 for those treated with the mixture and on day 11 for the others. Lactations were induced (minimum of 4.5 kg of milk/day) in five of six heifers and two of six cows by the mixture and in six of six heifers and three of six cows for estradiol benzoate. Milk production was 44% of herdmates in the 16 induced lactations. Cows on the single treatment had lower production than the other three groups. Ovarian status, cycling, cystic, or static, was affected adversely in 5 of 16 animals induced successfully. Two of the 16, both heifers, carried calves to term following induction. The transition to normal composition of milk was slower for single than double treatment. Lactose increased slowly to normal over the 1st wk of milking while protein decreased slowly. Estrogen and progesterone in milk of induced cows were approximately twice as concentrated as in normal post-parturient cows, probably because milk production was halved.     

Development of a fermented goat's milk containing probiotic bacteria

A set-type fermented milk manufactured from goat's milk was developed. Optimal curd tension was achieved by supplementation of milk with skim milk powder and whey protein concentrate (WPC). Milk was fermented employing a commercial probiotic starter culture (ABT-2), which contained Streptococcus thermophilus ST-20Y, Lactobacillus acidophilus LA-5, and Bifidobacterium BB-12. Supplementation of milk with 3% WPC reduced fermentation time by 2 h due to the increase in viable counts of S. thermophilus and Bifidobacterium by 0.3 and 0.7 log units, respectively. Addition of WPC increased the protein content (1%) as well as potassium and magnesium content (0.3 and 0.02 g kg⁻¹, respectively). Increase of the protein content led to an increase in the apparent viscosity and gel firmness of the product, and at the same time whey syneresis was reduced. As a consequence, the product received a high score for appearance, taste, aroma, texture and overall acceptance.     

Influence of Protein and Fat Contents of Ultrafiltered Milk on Rheological Properties of Gels Formed by Chymosin

Enzymatically induced gelation of milk and milk concentrated by ultrafiltration was studied with a curd firmness tester which measured transmission of oscillatory deformations through the product. Scott Blair's relationship between pseudo modulus of rigidity (G) and time after clotting (t), G = G∞ e^—_λ/t described the gelation of ultrafiltered retentates satisfactorily. Maximum gel firmness G∞, and _λ, which is the time to reach a firmness of G∞/e, were affected substantially by protein content. The effect of protein content on rates of firming of gels were evaluated at different stages of gel formation. At the onset of gelation, rates increased up to protein content 90 g kg^—1 but decreased at higher protein concentrations. At a later stage, this decrease disappeared and rates correlated with protein content. Those observations are discussed relative to effects of protein content on proteolytic and aggregation phases of milk clotting.Times from clotting to point of cutting curd were affected significantly by slight increases in protein above that in unconcentrated milk, but clotting times were not affected. Variations in fat content had inconsequential effects on test measures.     

Casein Composition of Cow's Milk of Different Chymosin Coagulation Properties

Five good and four poor chymosincoagulating individual cow milk samples were analyzed for casein composition using hydroxyapatite chromatography and polyacrylamide gel electrophoresis to establish possible relationships between casein fractions and differences in coagulation properties.Samples exhibited wide variation in casein composition. Poor chymosincoagulating milk had higher content of γ- and degraded caseins and lower κ- and β-caseins than the good-coagulating milk. One milk sample that did not coagulate 30 min after chymosin addition had low α_S-casein concentration and an additional major casein fragment (not identified). A substantial peak representing unidentified minor caseins was apparent in a poorcoagulating milk sample, which coagulated early but whose coagulum did not become firm in 30 min. Excluding the nonclotting sample, less variability was observed in α_S-casein concentrations than in the other casein components.     

Role of Exogenous Oxytocin in Eliciting Milk Ejection in Dairy Cows

The efficacy of small doses of exogenous oxytocin in eliciting milk ejection was measured in eight lactating Holstein cows. Treatments comprised intravenous administration of .02, .05, .10, and .30 IU of oxytocin. The smallest dose of oxytocin that induced milk ejection in all cows was .10 IU. Administration of .10 and .30 IU produced greater peak rates of milk flow and shorter milking times. Ranges for these two variables were 2.9 to 3.9 kg/min and 4.8 to 5.6 min. Variations in milk yield (8.2 to 9.1 kg/milking) were not significant, but more fat was harvested after administration of the two higher doses. Differences in peak concentrations of serum oxytocin were 17.7 to 21.2 μU/ml. Differences in latent time before peak concentration was reached were 1.3 to 4.0 min.     

Manufacture of Cheddar cheese from high-pressure-treated whole milk

The cheese-making characteristics of high-pressure (HP)-treated milk were examined. The rennet coagulation time of pasteurised milk decreased after HP treatment at 400 MPa but increased after treatment at 600 MPa. The L-value (whiteness) of milk decreased directly after HP treatment but, over the course of coagulation, whiteness of HP-treated milk increased to the same level as in the control. Cheddar cheese was then manufactured from raw whole milk or whole milk treated by high-pressure (HP) at 400 MPa (HP400) or 600 MPa (HP600) for 10 min at 20 °C. HP treatment of raw milk at 600 MPa resulted in a 3.66 log reduction in the initial counts of non-starter lactic acid bacteria (NSLAB), decreased protein and fat content, as well as a lower pH compared to the control. Furthermore, higher treatment pressures resulted in increased incorporation of β-lactoglobulin into the cheese curd, with parallel increases in yield by 1.23% and 7.78% for HP400 and HP600 cheeses, respectively. Overall, this study showed that the effects of HP treatment on milk proteins increased rennet coagulation times and changes in cheese composition at day 1.Industrial relevanceHigh-pressure treatment is a novel technology which has been applied to a number of commercial food products. In this study, HP-induced changes in milk proteins resulted in increased cheese yields and increased cheese whiteness. In addition, HP treatment significantly reduced the microflora of raw milk cheese. Those attributes could be of interest for both industry and consumer.