Capillary electrophoresis using a hydrophilically coated capillary and a low pH buffer containing urea has been used to follow the proteolytic action of plasmin and chymosin on isolated casein fractions, whole casein and individual milk samples selected on the basis of their genetic variants. Several of the main casein breakdown products were identified. These included, among others, γ1-casein (CN) A1, γ1-CN A2, γ1-CN B, γ1-CN C, γ2-CN A, γ1-CN A3, γ2-CN B, γ3-CN A and γ3-CN B, as well as proteose peptones, arising from the action of plasmin on the different genetic variants of β-CN. αs1-I-CN and αs1-CN f(1-23) from αs1-CN, and para-κ-CN and caseinomacropeptide from κ-CN produced by chymosin action were also separated. The knowledge of their migration times provided information on the extent and origin of casein hydrolysis in both milk and cheese, as found in samples of proteolysed milk or fresh cheese.

In dairy cows the period of transition from late gestation to early lactation is recognized as inducing considerable metabolic adaptation. The aim of this study was to analyse modifications in serum protein values occurring during the dry and the transition period and during lactation in a group of five Holstein cows of high average milk production. For all subjects, selected on the basis of their pregnancy status, blood samples were collected at different physiological phases: dry period (−60, −30 d to calving), transition period (almost 7 d to calving, 7 d after calving), and lactation (weeks 2, 5 and 15 after calving), for a total of eight blood samples for each cow. On each blood sample total proteins and electrophoresis analysis were performed. On the data obtained, normally distributed (P<0·05, Kolmogorov-Smirnov's test), one-way Repeated Measure Analysis of Variance (ANOVA), was applied to evaluate the influence of different stages of gestation and lactation on the considered parameters. Results showed a significant effect on total proteins, α1-globulins, β-globulins, γ-globulins and albumin/globulin ratio. Most of the detected modifications were related to the transition from gestation to lactation, indicating that it is a period of great metabolic stress for cows. On the basis of the obtained results we can affirm that the pattern of serum protein fraction rn could give information about dehydration, plasma volume expansion and hepatic function occurring during the peripartum period in dairy cows.

The main compounds responsible for the aroma of bovine, ovine, caprine and water buffalo freshly secreted milk have been identified by means of a gas chromatography–olfactometry technique. Of the fourteen odour-active volatile compounds detected, eight were present in all milks studied. Ethylbutanoate and ethylhexanoate (fruit-like aroma), among the neutral odorants, were the major contributors to the odour of bovine, ovine and caprine milk. The aroma of water buffalo milk was less dependent on ethylhexanoate and was also due to l-octen-3-ol (aroma of raw mushrooms), nonanal (freshly cut grass), indole (stable, animals) and an unidentified constituent, characterized by a retention index of 828, with a typical odour of warm milk and/or smoked cheese. Other aroma components were specific for some types of milk, contributing to the complexity and richness of flavour.

The objective was to compare the prevalence of subclinical mastitis (SM) and of udder pathogens in 60 Swiss organic (OP) and 60 conventional production systems (CP). Cows (n=970) were studied for SM prevalence and udder pathogens at median 31 d and 102 d post partum. Cows showing a [ges ]1+ positive California Mastitis Test (CMT) in at least one quarter were considered to have SM. Cow-level prevalences of SM for visits at 31 d and 102 d post partum (39% and 40% in OP and 34% and 35% in CP) were similar, but quarter-level prevalences of SM were higher (P<0·02) in OP than CP (15% and 18% in OP and 12% and 15% in CP). Median somatic cell counts in milk at 31 d post partum were higher (P<0·05) in OP than CP cows (43000 and 28000 cells/ml, respectively), but were similar at 102 d post partum in OP and CP cows (45000 and 38000 cells/ml, respectively). In milk samples from quarters showing a CMT reaction [ges ]2+ the prevalences of coagulase negative staphylococci were lower (P<0·05) at 102 d post partum, whereas prevalences of non-agalactiae streptococci were higher (P<0·05) in OP than in CP cows at 31 d and 102 d post partum. In conclusion, under Swiss conditions, subclinical mastitis is a greater problem in organic than in conventional production systems, but differences are not marked.

An experiment was conducted to study the seasonal changes in the fatty acid profile of milk from yaks (Bos grunniens) when kept at altitudes of 3000 m above sea level (a.s.l.) and higher. Data and samples were collected in summer (July), autumn (September), winter (November) and spring (March) from ten lactating yaks (four in spring). The yaks grazed pastures adjacent to the farm building throughout the year. In spring only they received 0·6 kg crop by-products per day (dry matter basis). Fresh alpine grasses, available in summer and autumn, showed high concentrations of α-linolenic acid (46–51 g/100 g lipids) compared with the dry, yellow vegetation of winter and spring (16 g/100 g lipids). In autumn and summer, the milk fat had higher concentrations of polyunsaturated fatty acids than in winter. These polyunsaturated fatty acids were comprised of vaccenic acid, rumenic acid and α-linolenic acid, which are all considered beneficial to human health. The rare fatty acid, γ-linolenic acid, was also detected in yak milk, especially in the milk obtained in spring. The results suggest that yak milk, which is the most important basic food of the Tibetan herders, has the most favourable fatty acid profile when yaks grazed green pasture, which also corresponds to the period of highest milk production.

The cheesemaking properties of recombinant chymosin from Escherichia coli were compared with those of standard rennet in parallel trials with three single strain starters. For each pair of cheeses the cheesemaking characteristics, recoveries of milk solids in the curds and compositions were similar. The rates and types of proteolysis and texture development during ripening between cheeses made with the two coagulants were not significantly different.

1. The sensitivity to calcium of the caseinate-phosphate complex of milk heated at 120°C., increases to an early maximum and thereafter steadily declines, apparently in consequence of parallel changes in the sensitivity of its caseinate constituent.

2. A simultaneously developed capacity of the heated complex to bind additional colloidal phosphate appears to be unrelated to these changes in casein sensitivity.

3. Colloidal phosphate content is confirmed as a factor in the heat coagulation of milk.

4. Heat-developed acidity contributes to the heat coagulation of milk primarily by increasing hydrogen-ion concentration and only slightly, if at all, through release of calcium ions from insoluble combinations.

5. Neither serum proteins nor protein-lactose combinations play any significant part in the heat coagulation of milk.

6. The bearing of these results on the heat coagulation of milk in general and on the operation of certain high-temperature stabilization treatments is discussed.

Two types of artificial neural networks, multilayer perceptron (MLP) and self-organizing feature map (SOM) were used to detect mastitis by automatic milking systems (AMS) using a new mastitis indicator that combined two previously reported indicators based on higher electrical conductivity (EC) and lower quarter yield (QY). Four MLPs with four combinations of inputs were developed to detect infected quarters. One input combination involved principal components (PC) adopted for addressing multi-collinearity in the data. The PC-based MLP model was superior to other non-PC-based models in terms of less complexity and higher predictive accuracy. The overall correct classification rate (CCR), sensitivity and specificity of this model were 90·74%, 86·90% and 91·36%, respectively. The SOM detected the stage of progression of mastitis in a quarter within the mastitis spectrum and revealed that quarters form three clusters: healthy, moderately ill and severely ill. The clusters were validated using k-means clustering, ANOVA and least significant difference. Clusters reflected the characteristics of healthy and subclinical and clinical mastitis, respectively. We conclude that the PC based model based on EC and QY can be used in AMS to detect mastitis with high accuracy and that the SOM model can be used to monitor the health status of the herd for early intervention and possible treatment.

1. Certain starter cultures have been found to be inhibited in the presence of milk peroxidase.

2. A high correlation has been established between the inactivation of milk peroxidase and of the inhibitory action by use of critical values of any of the following: temperature, pH, concentration of either sodium azide or hydrogen peroxide.

3. The inhibitory action can be overcome by the addition of certain reducing substances such as cysteine and sodium hydrosulphite.

4. The inhibitory action is exhibited by the corresponding separated milk and whey.

5. It is suggested that this inhibition may be due to the formation of a specific inhibitory oxidation product having a quinonoid structure.

6. The addition of 1–2% of pasteurized milk or whey to autoclaved milk caused total inhibition, whereas such additions to milk heated to 190° F. showed no such effect. This is attributed to the presence of SH groups in the latter milk capable of reducing the oxidation product.

7. It is suggested that the inhibition due to peroxidase is identical with that ascribed to lactenin 2, and that the inhibition described in our previous paper as due to ‘agglutinin’ is identical with that ascribed to lactenin 1.

Two experiments were conducted to investigate possible changes of milking-related oxytocin release (Expt 1) and of intramammary pressure and milking characteristics (Expt 2) throughout entire lactations in German Braunvieh dairy cows. Mean oxytocin concentrations after stimulation at onset of milking increased from 18·3 ± 15·9 to 30·7 ± 24·1 pg/ml in Expt 1 and decreased from 23·9 ± 17·6 to 15·4 ± 9·1 pg/ml in Expt 2, respectively, but remained above the level necessary to elicit complete milk ejection in both trials. Premilking baseline intramammary pressure had its maximum in early lactation until about month 4 and then decreased to ∼50% of its initial level. Ejection pressure followed a similar pattern, but dropped only to ∼75% of its maximum. This was due to the constant elevation of pressure increase, reaching its highest level in late lactation. Time from commencement of stimulation until maximum pressure exceeded 1 min in almost all instances even in early lactation and increased throughout lactation. Despite the normal decrease of milk yield average milk flow fell only slightly while maximum flow rate remained almost constant. Pressure increase, milk yield and milk flow were not different after 1 min and after extended stimulation. Thus there were no indications of a decreasing sensitivity of the milk ejection reflex during lactation, and milking characteristics were positively affected by intense teat stimulation. Suggestions for practical dairying are made.

For 17 phages active against Streptococcus cremoris, Str. lactis and Str. lactis subsp. diacetylactis, the killing efficiency of pasteurization (log No/N) at 72°C for 15 s in skim-milk showed large variations from > 6 to 0; the efficienty of killing during spray-drying ranged from 3.7 to 0.2 and phages survived well storage of milk powder at room temperature. Destruction in a heat exchanger was found to be greater than that calculated from biphasic curves obtained by heating phages in sealed ampoules. No relationship was established between lytic classification of phages and their heat resistance.

The aim was to select potentially probiotic lactobacilli from 88 strains isolated from unpasteurized milk and cheese products, and to incorporate these bacteria in a viable state into a soft cheese, without changing its quality. The survival of these bacteria was assessed in acidic and bile conditions, after freezing at −80 °C. Four strains from unpasteurized Camembert – two Lactobacillus plantarum strains and two Lb. paracasei/casei strains – were identified and typed by PCR and PFGE and were found to display potentially probiotic characteristics in addition to resistance to low pH and bile. These characteristics were resistance to lysozyme, adhesion to CACO-2 cells, antimicrobial effects against common foodborne pathogens (Listeria monocytogenes, Staphylococcus aureus, Salmonella spp., Escherichia coli), innocuity following the ingestion of high doses by mice and appropriate antibiotic susceptibility profiles. The potential of Lb. plantarum strain UCMA 3037 for incorporation into a soft cheese (Pont-l'Eveque registered designation of origin (RDO)) was investigated. This strain grew well and survived in sufficient numbers (more than 107 cfu/g throughout the shelf-life of the product) in the cheese. This strain did not change the quality score of the product until the best before date (75 days after manufacture). Thus, unpasteurized Camembert is a natural source of potentially probiotic lactobacilli, which could be used as an additive in the development of potentially probiotic soft cheeses. Further work is required to demonstrate the persistence and efficacy of these strains in the human host upon ingestion.

k-Casein is the main glycoprotein of cow's milk. Its polysaccharide part is O-glycosidically linked to threonine residue 133. It contains only 3 different sugars (Gal, GalNAc, NeuNAc), but a microheterogeneity has been detected at the sugar level. Two main polysaccharides have so far been characterized. The structure of the trisaccharide is NeuNAc α → 3 Gal β1 →3 GalNAc; the tetrasaccharide contains one additional sialic acid. The polysaccharide part of ovine k-casein resembles that of bovine k-casein, but contains also N-glycolyl neuraminic acid. Human k-casein contains 3 times more carbohydrate than bovine k-casein with 2 additional sugars, GlcNAc and Fuc. The various polysaccharide parts isolated from bovine colostrum k-caseinoglycopeptide are much more complex than those obtained from the normal glycopeptide, indicating an evolution of the sugar part as a function of time after parturition. Some aspects of the secondary structure of k-casein and the role of the sugar part are discussed. The carbohydrate moiety of another milk protein, human lactotransferrin, is also discussed briefly. It is comprised of 2 identical glycan groups, N-glycosidically linked to the protein, and quite different from the k-casein carbohydrate moiety.

A fraction derived from the membrane of milk fat globules (MFGM) was isolated from fresh raw cream. The adsorption of MFGM at the interface of oil-in-water emulsions was studied as well as the stability of the emulsions under different conditions (pH, temperature treatment) and the surface potential. The emulsions prepared with MFGM isolates were stable at neutral pH and were destabilized at low pH. By analysis of the protein adsorbed at the interface by polyacrylamide gel electrophoresis, we also determined that the MFGM that covered the surface of the oil-in-water emulsion could not be displaced by surfactants such as Tweens and Triton X-100 and was not affected by the presence of other proteins such as caseins and β-lactoglobulin added to the emulsion. A strong molecular interaction between the adsorbed MFGM, composed of phospholipid and protein, may exist at the interface, rendering the newly formed membrane very little affected by the presence of other surfactants. This MFGM-stabilized emulsion was characterized by a behaviour very different from that of emulsions stabilized by other milk proteins.

Cell-free extracts of both Lactobacillus bulgaricus and L. acidophilus demonstrated threonine aldolase activity, the end product of which was acetaldehyde, the major flavour compound of yoghurt. L. acidophilus also possessed an alcohol dehydrogenase activity capable of reducing acetaldehyde so that little yoghurt flavour was present in milks fermentation with this organism. Addition of threonine to fortified milk before fermentation with L. acidophilus increased acetaldehyde production and resulted in a well flavoured product similar to that of yoghurt made with L. bulgaricus. The contribution of these 2 enzymes to flavour production is discussed.

The effects of decreasing pH and micellar calcium concentrations of reconstituted skim milk and caseinate solution were studied by 1H and 17O NMR spectroscopy. The proton transverse relaxation rate 1/T2 of skim milk decreased as the pH decreased, reaching a minimum at pH 5·3. However, as the pH fell sodium caseinate solution showed a continuous increase in 1/T2, with no minimum. Analysis of proton relaxation as a function of the interpulse time in the CPMG (Carr-Purcell-Meiboom-Gill) sequence demonstrated that both the proton exchange mechanism and ‘bound’ water contributed to proton relaxation in skim milk. The study of 17O relaxation rate as a function of pH confirmed the change in protein hydration upon acidification. Increasing the amount of EDTA showed that the proton transverse relaxation rate of skim milk decreased until a plateau was reached when the micellar calcium was totally solubilized. With excess EDTA the relaxation rates of skim milk and caseinate solution were identical. A strong correlation was also found between the pH dependent relaxation rate and the solubilization of micellar phosphorus as detected by 31P NMR. Together, these results suggested that aggregation of caseins by calcium and colloidal calcium phosphate is mainly responsible for the excess hydration in skim milk micelles.

An objective test has been developed to help in assessing the value, as a method for determining the heat stability of milk, of the subjective test described by Davies & White (1966a). The objective test showed that coagulation time, as determined by the subjective test, is a reasonably accurate measure of the induction period that precedes the onset of initial rapid coagulation, but that coagulation proceeds in 2 different ways depending primarily on whether the coagulation of the milk is good (initial clots large) or poor (initial clots small). The latter finding confirms the view that in studying heat stability these 2 categories of milk should be examined separately.

Prolidase activity from cytoplasm of Lactococcus lactis subsp. cremoris (Streptococcus cremoris) AM2 was partially purified. The enzyme had Mr 42000 and optimum activity between pH 7·35 and 8·25 in citrate, phosphate and borate buffers while in a universal buffer system an optimum pH between 8·3 and 9·0 was observed. The activity was strongly inhibited by the chelating agents E.DTA, 1,10-phenanthroline and 8-hydroxyquinoline. Inhibition was also noted with dithio-threitol, N-ethylmaleimide and bacitracin. The enzyme was active on all amino-acylproline substrates tested except Gly-Pro and Gip-Pro and also showed activity against Pro-Pro. While most prolyl amino acids tested were not hydrolysed, hydrolysis was noted with Pro-Ala and Pro-Val. Km values of 20 mM and 10 mM were obtained with Phe-Pro and Met-Pro respectively; however, substrate inhibition was observed with Ile-Pro and Leu-Pro.

The formation of protein polymers in ultra-high-temperature (UHT)-treated milk during storage at various temperatures was examined by gel filtration. The extent of polymer formation was found to depend on both storage time and temperature. After some months of storage at the higher temperatures of 30 and 37°C, the extent of polymerization of the caseins and whey proteins due to reactions of the Maillard type was several times greater than the heat-induced changes resulting from the UHT processing itself. After storage for 6 months the following proportions of milk proteins were found to exist in the form of covalently bound polymers: 50% at 37°C, 40% at 30°C, 26% at 20°C and 21% at 4°C. In addition, further amounts of polymer were formed by disulphide bonding, the contribution of such polymers diminishing gradually in a temperature-dependent manner during storage due to continuing polymerization reactions. It appeared that αs1-casein may be preferentially involved in these polymerizations with β-casein reacting at a somewhat slower rate. Polymerization and associated reactions modifying molecular charge led to the expected alterations in electrophoretic mobility and a loss of definition in the bands due to the various protein components. Measurements of proteolytic breakdown indicated only very small increases in trichloroacetic acidsoluble fragments, suggesting that proteolysis was of minor significance.

The effect of the proportion of clover in the diet (200, 500 or 800 g/kg total dry matter (DM)) on milk production of cows housed indoors and fed on a mixture of perennial rye-grass and white clover was measured in mid (Expt I) and late (Expt II) lactation. Higher clover contents increased the nutritive value of the diets, resulting in increased energy and protein intakes. DM intakes of cows offered 500 or 800 g clover/kg DM diets ad lib. (Expt I and Expt II, Period 1) were not significantly different but were 11–17% greater (P<0·05) than intakes of cows fed on 200 g clover/kg total DM diets. Cows offered restricted allowances (Expt II, Period 2) had similar intakes irrespective of diet. In Expt I cows fed on 500 or 800 g clover/kg DM diets ad lib. produced 30 or 33% respectively more milk (P<0·05) than cows fed on 200 g clover/kg total DM diets. During Expt II, Period 1, cows fed on 500 or 800 g clover/kg DM diets ad lib. produced 18 or 16% more milk (P<0·05) respectively than cows given 200 g clover/kg total DM diets. In both these experiments the increased milk yields were due to increased intake and the higher nutritive value of the high clover diets. There was no difference in the feed conversion efficiencies of cows if maintenance energy requirements were taken into account. However, cows on restricted allowances (Expt II, Period 2) showed no significant difference in milk yield, indicating that the effect of increased nutritive value was very slight. There were no consistent effects on milk fat, protein or lactose concentrations. Concentrations of blood and milk urea increased as the clover content of the diet increased (Expt 1 only), and this was associated with increased milk non-protein N and a decreased ratio of casein N[ratio ]total N. Both trials indicated an optimum clover content in the diet for milk production of 600–700 g/kg total DM.