The primary structures of ovine α>s1-casein variants A, C and D (formerly called Welsh variant) were determined. Separation of variants from whole casein was achieved using a fast and reliable reversed-phase HPLC method. Extended structural characterization of the purified proteins using electrospray mass spectrometry, automated Edman degradation and peptide mapping by means of HPLC-fast atom bombardment-mass spectrometry demonstrated that the mature protein was a mixture of two molecular species that differed in the deletion of residues 141–148 and were therefore 199 and 191 residues long respectively. The 199 residue peptide chain, which accounted for ∼ 80% of the entire translated αsl-casein, was as long as its caprine and bovine counterparts, and had a 98 and 89 % degree of identity with those two proteins respectively. Nine serine residues (positions 12, 44, 46, 64 to 68 and 75) were fully phosphorylated in αsl-casein A, whereas Ser115 and Ser41 were phosphorylated by ∼ 50 and ∼ 20% respectively. The differences between the three genetic variants A, C and D were simple silent substitutions, which however involved the degree to which the protein was phosphorylated. Variant C differed from variant A in the substitution Ser13 →> Pro13 which determined the loss of the phosphate group on site 12 of the protein chain, SerP12→>Ser12. A further substitution, SerP68 →> Asn68 caused the disappearance of both phosphate groups in the phosphorylated residues Ser64 and Ser66 in variant D; in this last casein variant there was no evidence of phosphorylation at Ser41.

Yogurts are important dairy products that have known a rapid market growth over the past few decades. Industrial yogurt manufacture involves different processing steps. Among them, protein fortification of the milk base is elemental. It greatly enhances yogurt nutritional and functional properties and prevents syneresis, an undesirable yogurt textural defect. Protein enrichment can be achieved by either concentration process (evaporation under vacuum and membrane processing: reverse osmosis and/or ultrafiltration) or by addition of dairy ingredients. Traditionally, skim milk powder (SMP) is used to enrich the milk base before fermentation. However, increased quality and availability of other dairy ingredients such as milk protein isolates (MPI), milk protein concentrates (MPC) whey protein isolates (WPI) and concentrates (WPC), micellar casein (MC) and caseinates have promoted their use as alternatives to SMP. Substituting different dry ingredients for skim milk powder in yogurt making affects the yogurt mix protein composition and subsequent textural and sensorial properties. This review focuses on various type of milk protein used for fortification purposes and their influence on these properties.

The milk-clotting and proteolytic activities of rennet, bovine pepsin and porcine pepsin were compared. The milk-clotting activity of porcine pepsin was extremely pH-dependent around pH 6·6 and coagulation did not occur above pH 6·68. The clotting activity of bovine pepsin was slightly more dependent on pH than that of rennet but no rapid drop-off in activity occurred as with porcine pepsin. Temperature influenced the clotting activity of rennet and bovine pepsin similarly but the behaviour of porcine pepsin was markedly different.

For equal milk-clotting activity, the proteolytic activities of rennet and bovine pepsin were approximately equal and substantially lower than that of porcine pepsin. Electrophoretic examination showed that the proteolysis products of rennet and bovine pepsin were similar and quite different from those produced by porcine pepsin.

The suitability of bovine pepsin as a rennet substitute is discussed.

A new sampling method for the collection of fore and hind milk from the sow has been developed which resembles normal milk removal by the piglet, yet overcomes the difficulties of collecting milk that is available for only a short time (10–20 s) at each let-down. Samples of hind milk were collected immediately after the completion of a successful sucking, while the fore milk was collected at the beginning of the next natural let-down. Modification of existing assays for fat, protein and lactose was required to provide rapid analysis of the small volumes (< 0·5 ml) of milk collected using this procedure, and these methods were validated by comparison with reference methods. Total solids and the concentration of fat in whole milk, and protein and lactose in skim milk, were measured in fore and hind milk collected from 4, 20, 12 and 12 sows respectively, 6–11 d post partum. For fore milk, the results (mean ± SD (n)) were: total solids, 199·9±9·9 g/1 (8); fat, 96·7 ± 13·9 g/1 (41), protein, 47·7±4·5 g/1 (27) and lactose, 58·3 + 4±0 g/1 (27). In hind milk, there was a significant increase in the concentration of fat (15·3 g/1, p < 0·001, n = 41) which was reflected by a significant increase in total solids (14·7 g/1, P < 0·001, n = 8) and calculated energy (511 kj/l, P < 0·001, n = 27), but there was no significant difference in the concentration of either protein or lactose. This increase in milk fat during milk letdown is in contrast to the results of most previous studies in the sow and is discussed in terms of the possible mechanisms that may cause the concentration of fat to increase as milk is removed from the mammary gland.

The folate in cow's milk was strongly and specifically bound to a minor whey protein (FP), forming a complex of primary M of about 38000, but exhibiting concentration-dependent reversible aggregation. The binding protein was present in excess, and the milk had the capacity to bind about 50 μg added folic acid/l. An enriched concentrate of FP was prepared by ammonium sulphate fractionation—FP was precipitated at between 50 and 60% saturation—and further purified by chromatography in DEAE-cellulose and filtration in Sephadex gel G150. Its identity as a distinct minor whey protein was confirmed by comparative starch gel electrophoresis at various pH values.

Some properties of the protein are described, and its physiological significance discussed.

Casein breakdown and free fatty acids formation were studied in ten batches of La Serena cheese throughout a 60 d ripening period. A rapid degradation of αs1-casein occurred, with only 18·6% residual αs1-casein being detected in 60 d-old cheese, whereas 49·5% β-casein remained unattacked. High pH values, low NaCl-in-moisture concentrations and high moisture contents enhanced proteolysis. Lac-tobacilli and yeasts were the only microbial groups which significantly influenced αs1-casein breakdown, and only yeasts affected β-casein hydrolysis. No significant effect of chemical characteristics or microbial groups on FFA levels was detected, an average level of 12·7 mequiv./100 g fat being recorded for 60 d cheese. Rheological characteristics of the cheese reached their minimum values after 15–30 d of ripening and were dependent on αs1- and β-casein breakdown, pH value, NaCl-in-moisture concentration and moisture content.

Measurements have been made of the coagulation times (CT) of concentrated and diluted milks when treated with chymosin. After dilution, milks showed a large increase in CT: conversely, concentrated milks showed a similar CT to unconcentrated milk. The results are discussed in terms of a mechanism which reduces to a form similar to that proposed by Holter (1932).

The streptococci occurring in raw and pasteurized milks of varying purity have been investigated. The organisms were placed in five main groups by testing for the hydrolysis of arginine, growth at 45 and at 8–12° C, survival in milk at 63° C. for 30 min. (measured by a semi-quantitative plating method), action on litmus milk, and the ability to form CO2 from glucose (tested by a cultural method). The groups thus distinguished are:

(1) Organisms of bovine mastitis.

(2) Str. bovis and Str. thermophilus. A clear-cut separation of these species was not obtained by tests for growth at 50° C, the hydrolysis of aesculin, or the fermentation of maltose or salicin; but, with suitable precautions, action on both maltose and salicin provides a differentiation. Certain strains of Str. bovis differ from the typical form of that species in their ability to proliferate over a wider range of temperature and in higher concentrations of salt and methylene blue and in their failure to split aesculin.

(3) Str. lactis and Str. cremoris, which were differentiated by tests for the hydrolysis of arginine and ability to grow at 40° C.

(4) Str.faecalis and its varieties. Many of the strains are non-haemolytic but otherwise indistinguishable from Str. durans. The formation of acetoin from citrate and from glucose are common but not universal properties in the enterococcus group.

(5) Heterofermentative streptococci. The majority of the strains were identified as Str. kefir. Str. citrovorus, which was isolated only occasionally, was distinguished by its low maximum temperature (under 37° C), weak resistance to heat and by fermentation reactions.

Raw milk was found to contain Str. kefir, Str. lactis and the mastitis organisms more frequently than the other species. Str. thermophilus and Str. bovis were the dominant forms in freshly pasteurized milk. Pasteurized milk held at 10–22° C. until it became tainted yielded Str. kefir and Str. faecalis and, at the higher temperatures, Str. bovis and Str. thermophilus.

The properties of adsorbed films of αs1-, β- and κ-casein and sodium caseinate are compared at the planar oil–water interface, at the surface of emulsion droplets, and at the surface of polystyrene latex particles. Attention is directed towards understanding (i) the relationship between the molecular characteristics of the caseins and their surface activities and surface viscosities at the oil–water interface, (ii) the nature of competitive adsorption between the caseins (particularly αs1- and β-), and (iii) the effect of Na and Ca ion concentrations on electrophoretic mobilities and flocculation behaviour.

Ten-day periods of water restriction followed by 2 h ad lib. water caused radical changes in camel milk. The changes were different from those reported in other mammals. At the end of each dehydration period there were increased water percentages in the milk when compared with milk of hydrated camels. The content of fat, lactose and protein declined. Concentrations of Na, K, phosphate and chloride in milk increased while Ca and Mg concentration declined. The urea concentration of the milk was unchanged. The young suckling calves showed no signs of stress during the dehydration periods. It was concluded that the milk of dehydrated camels is an excellent human food in arid areas, as the water and salt content are high while the nutritive value remains good.

Micelles of milk heated at 95 °C for 10 min or autoclaved at 121·7 °C for 15 min were observed by electron microscopy to bear filamentous appendages, whereas micelles from raw milk had relatively smooth contours with no appendages. Experiments suggested that the appendages were composed of denatured β-lactoglobulin and showed that their development was sensitive to sulphydryl blocking agents. Firmness and syneresis of yoghurt were associated with the extent to which micelles coalesced during fermentation; the appendages of micelles in heated milk appeared to inhibit coalescence giving rise to a firmer curd with a lower tendency to syneresis.

The aim of this work was to study the spin–spin (T2) relaxation components of one hard cheese and three soft cheeses to characterize fat and water states. NMR signals were measured at 6 °C with a 0·47 T NMR device. The transverse relaxation decay was fitted using the Marquardt method. The T2 relaxometric behaviour of the cheeses under consideration was characterized by four relaxation components. To understand the chemical composition of each NMR component, we studied anhydrous milk fat extracted from each cheese analysed. At 6 °C, the fat was 60% crystalline. In cheese, the solid fat was found mainly in the shorter relaxation component with a T2 of 17 μs. The intensity of the NMR relaxation with a T2 > 1 ms was explained by the amount of water, liquid fat and proteins, and the associated relaxation time varied as a function of the process used. The composition of each relaxation component was confirmed by the temperature effect and the influence of the fat content on the NMR cheese signal. NMR relaxometry was able to provide information on water behaviour (i.e. the quantity and level of interactions with proteins) and on the solid[ratio ]liquid ratio of anhydrous milk fat in the cheese.

Cheese of the Swiss (Emmenthaler, Gruyère, Danish-Swiss) type is popular on the continent of Europe and in America, but severe losses in ripening are caused by contamination with anaerobic sporeformers. Active gas production by the clostridia during ripening causes distension and spoiling of the cheese which becomes unmarketable.

Groups of lactating cows and heifers were milked four times daily in two diagonally opposed glands for 4 weeks, and the effects on milk yield studied relative to twice-daily milked glands as controls. Mammary enzyme activities, in vitro synthesis rates of milk constituents and histological scoring were determined in mammary biopsy samples obtained at the end of this period. These were used for assessment of mammary function. Frequent milking increased milk yield only in the treated glands, the contralateral control glands continuing to decline in yield at ~ 2%/week. There was no significant difference in response between cows and heifers; the mean increase in yield was 10·4%. The rate of decline in milk yield tended to decrease with frequent milking, to ~ 1%/week. Consequently the yield of the treated glands continued to be elevated above that of the controls for some time after reversion to overall twice daily milking. Milk protein content was increased slightly by frequent milking. Mammary enzyme activities were ~ 18% higher in the treated glands than in the controls. Synthesis rates of lactose, casein and total protein were unaffected by milking frequency, but were all lower in the gland selected for the second biopsy, reflecting the reduction in milk yield caused by the first biopsy. DNA synthesis was increased by milking frequency, as were the size and number of epithelial cells in histological sections.

A new polymerase chain reaction (PCR)-based method was developed to detect cows' milk in goat cheese. This method is based on mitochondrial DNA (mtDNA) control region sequence variations. DNA extractions from 150 mg of cheese were carried out using a spin column-based method. Subsequent PCR amplifications of DNA were performed with cow specific primers, demonstrating the ability to detect cows' milk in a variety of cheeses. This simple approach provides high quality DNA, and is shown to be very sensitive, with a detection limit of less than 0·1% of cows' milk. Analysis of an agarose gel digital image allows a rough estimation of the percentage of cows' milk used in adulteration.

Yeast extract was fractionated on Sephadex G-25 into 7 fractions. The fraction most stimulatory to the growth of Streptococcus lactis C10 contained over 70% of the amino N present in yeast extract and consisted of a wide variety of free amino acids and a small amount of peptide material. Examination of possible replacement factors for this fraction revealed that the amino-acid material present was largely responsible for the stimulation of Str. lactis C10. Purine and pyrimidine bases and inorganic constituents also contributed to the stimulation. In addition, yeast extract contained a component which decomposed H2O2, a metabolite which accumulates in the growth medium under aerobic conditions and inhibits growth. The nature of the stimulation was studied by isolating slow and fast acid-producing colonies of Str. lactis C10. It appeared that yeast extract and other amino-acid supplements prevented an observed inhibition of the growth of the slow variants below pH 6·0, apparently by satisfying a nutritional deficiency caused by a drop in pH.

Reduction of the level of colloidal calcium phosphate (CCP) progressively increased the heat stability of milk at pH values <~7·0 and increased the pH of maximum stability. Removal of 40% CCP also stabilized the system at the pH of minimum stability, but removal of ≥60% CCP rendered milk very unstable at pH values >7·2, an effect not offset by a 4-fold increase in κ-casein concentration. Doubling CCP had a slight destabilizing effect in the pH range 6·5–7·5.

Addition of β-lactoglobulin to serum protein-free casein micelles had a marked destabilizing effect at pH values > ~6·8, but increased stability in the pH range 6·4–6·8. β-Lactoglobulin had a similar and more apparent effect on the heat stability of Na caseinate dissolved in milk diffusate.

It is suggested that rather than being a stabilizing factor responsible for the maximum in the heat stability-pH curve, the true effect of β-lactoglobulin is to shift the curve to more acid pH values (reason unknown) and to sensitize the caseinate system to heat-induced Ca phosphate precipitation at pH values > ~7·0. Low stability at ~pH 7·0 introduces an apparent maximum in the heat stability-pH curve at ~pH 6·8, but this has no independent existence. At pH values >7·2, increased protein charge more than off-sets the influence of heat-precipitated CCP and stability again increases in micellar but not in soluble casein systems.

Experiments were undertaken to validate a method (using adrenaline injection) for determination of the size of cisternal and alveolar compartments in the udder, to use this method to determine the pattern of milk accumulation in the udder over time and to determine the relationship between the size of the alveolar and cisternal compartments and tolerance of once daily milking. Cows received intrajugular injections of adrenaline (3 mg) immediately before milking, to block milk ejection and allow harvesting of the cisternal milk fraction. This was followed by removal of the alveolar fraction 30 min later after intrajugular oxytocin (5 i.u.) injection. Results obtained were similar to those obtained by catheter drainage. The alveolar compartment was 90% full at 16 h post milking while the cisternal compartment filled more slowly and was only 70% full at 24 h post milking. At full capacity (measured at 40 h), the volumes of milk contained in the cisternal and alveolar compartments were similar. In a further experiment involving identical twin cows, it was shown that the greater the degree of filling of the cisternal compartment at 24 h, the lower was the production loss on once daily milking. This suggests that the freedom of the alveoli to drain was an important factor in the production loss on once daily milking. Although there were significant correlations within twin sets for milk yield and the size of udder compartments, the relationship within twin sets for yield loss on once daily milking was not statistically significant.