The importance of elevated concentrations of oxytocin (OT) during the entire milking period was investigated in seven primiparous dairy cows with inherent disturbed milk ejection and in sixteen healthy cows with disturbed milk ejection induced by placing them in an operating theatre. Disturbance of milk removal in both groups has previously been demonstrated to be exclusively due to central blockage of the expected OT release in response to teat stimulation and milking. However, milk ejection can be induced by exogenous OT. OT (0·2 i.u.) was injected i.v. before milking and 49 ± 6% of the total milk was removed. When plasma OT decreased, milk flow stopped. In response to a second and third injection of 0·2 i.u. OT, 30 ± 4 and 7 ± 2% of the milk were removed respectively. The remaining milk was removed with 10 i.u. OT. The lag time from injection of OT to the start of milk flow was inversely correlated with the amount of milk actually removed in response to the OT injection. If 0·2 i.u. OT was injected during intramammary pressure (IMP) recording, IMP immediately increased to its maximum value. After 2·5 ± 0·3 min, IMP decreased to an intermediate IMP (between preinjection and maximum IMP). After two additional injections of 0·2 i.u. OT and after injection of 0·5 i.u. OT, IMP increased to a similar maximum. However, after injection of 0·5 i.u. OT, maximum IMP lasted longer (2·9 ± 0·3 min; P < 0·05) than after injections of 0·2 i.u. If OT was continuously infused (0·15 i.u./min) during milking, milk flow lasted until the udder was completely emptied. IMP increased during OT infusion to a maximum which remained stable until infusion was stopped after 10 min. The same IMP maximum was reached after the first individual OT injection (0·2 i.u.), but when plasma OT decreased towards basal concentrations, milk flow ceased and IMP decreased to an intermediate level. Thus continuously elevated OT concentrations such as those during infusion or during normal milking are necessary for complete milk removal.

The development of structure in Cheddar cheese was followed using electron microscopy, of both thin sections and freeze-etched preparations. The casein micelles, at first separate, were seen to aggregate into a network, coalesce and finally form a granular mass. The fat globules, also separate at first, were gradually forced into clumps as a result of shrinkage of the casein network and incomplete fusion took place during ripening. Starter bacteria were seen trapped in the casein near the fat–casein interface, which was shown to be the region of highest water content in the mature cheese.

The gross energy content of the milk of the cow, sow, ewe, goat and of human milk has been determined by bomb calorimeter, and compared with results obtained using conversion factors and chemical analysis.

Correction for non-protein nitrogen content led to values agreeing within 0·5–1·0% for the milk of all species. For human colostrum other factors may be involved, and a closer estimate of gross energy value of human milk was obtained by treating all nitrogenous constituents as being protein.

Nutritional interventions are now recommended as strategies to delay Alzheimer's disease (AD) progression. The present study evaluated the neuroprotective effect (anti-inflammation) of lactic acid bacteria (either Lactobacillus fermentum LAB9 or L. casei LABPC) fermented cow's milk (CM) against lipopolysaccharide (LPS)-activated microglial BV2 cells in vitro. The ability of CM-LAB in attenuating memory deficit in LPS-induced mice was also investigated. ICR mice were orally administered with CM-LAB for 28 d before induction of neuroinflammation by LPS. Learning and memory behaviour were assessed using the Morris Water Maze Test. Brain tissues were homogenised for measurement of acetylcholinesterase (AChE), antioxidative, lipid peroxidation (malondialdehyde (MDA)) and nitrosative stress (NO) parameters. Serum was collected for cytokine analysis. CM-LAB9 and CM-LABPC significantly (P < 0·05) decreased NO level but did not affect CD40 expression in vitro. CM-LAB attenuated LPS-induced memory deficit in mice. This was accompanied by significant (P < 0·05) increment of antioxidants (SOD, GSH, GPx) and reduction of MDA, AChE and also pro-inflammatory cytokines. Unfermented cow's milk (UCM) yielded greater cytokine lowering effect than CM-LAB. The present findings suggest that attenuation of LPS-induced neuroinflamation and memory deficit by CM-LAB could be mediated via anti-inflammation through inhibition of AChE and antioxidative activities.

The modulating effect of bovine milk casein components and their digests on the proliferative responses of mouse spleen lymphocytes and rabbit Peyer's patch cells induced or not induced by mitogens has been studied with a colorimetric assay using 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide. All the casein components and their digests tested had little mitogenic effect on the proliferative responses of mouse spleen lymphocytes and rabbit Peyer's patch cells. Intact κ-casein significantly inhibited the proliferative responses of mouse spleen lymphocytes and Peyer's patch cells induced by mitogens such as lipopolysaccharide from Salmonella typhimurium, concanavalin A, phytohaemagglutinin and pokeweed mitogen. In contrast, intact αsl-casein and β-casein had little effect. κ-Casein had an inhibitory effect after digesti on by pancreatin or trypsin, but not after pepsin or chymotrypsin digestion. Both pancreatin and trypsin digests of αsl-casein and -casein significantly inhibited the proliferative responses of mouse spleen lymphocytes and rabbit Peyer's patch cells induced by mitogens, whereas pepsin and chymotrypsin digests of both caseins were without effect. Moreover, the trypsin digest of each casein component had an inhibitory effect on mouse spleen lymphocyte proliferation in the absence of mitogen. Since trypsin is a major proteinase in pancreatin, the substrate specificity of trypsin seems to be important for the formation of the inhibitory peptides from casein components. These observations suggest that intact κ-casein and some peptides formed from milk casein components by the action of trypsin may suppress the immune responsiveness of neonates.

Transglutaminase (TGase) is an enzyme that cross-links many proteins, including milk proteins. In this study, the effects of TGase on some physico-chemical properties of milk were studied. TGase-treated milk was not coagulable by rennet, which was due to failure of the primary (enzymic) stage of rennet action rather than the non-enzymic secondary phase. Dissociation of TGase-treated casein micelles by urea or sodium citrate or removal of colloidal calcium phosphate by acidification and dialysis was reduced, presumably due to the formation of cross-links between the caseins. Casein micelles in TGase-treated milks were also resistant to high pressure treatment and to hydrolysis by plasmin. Results of the present study show that milk proteins are fundamentally modified by the action of TGase, which may have applications in the manufacture of functional proteins for use as novel food ingredients.

The rheological properties of rennet-induced skim milk gels at 30 °C are compared with casein gels formed by acidification to pH 4·6 at 2 °C and subsequent heating to 30 °C. Both types of gels are viscoelastic. However, the gels formed by rennet action are relatively more viscous over time scales longer than 1 s. This implies that on average the protein-protein bonds in these gels are relatively more mobile than in the acid casein gels. It is thought that this is primarily due to differences in the structure of the casein particles and in the type of the main interaction forces in and between the casein molecules and particles.

The ferric thiocyanate method of estimating peroxide in fats has been modified to give greater sensitivity and greater stability of the reagent. As the 96% acetone used by Chapman & McFarlane (3)and Lips et al.(4) only dissolves about 0.5% butterfat, the sensitivity of the test was increased by using, as solvent, the mixture of benzene and methanol used by Bolland et al. (6) in the determination of rubber peroxides, which dissolves at least 10% butterfat. Instead of using a prepared reagent which gradually deteriorates, the ferrous salt and thiocyanate are added in aqueous solution during the test, and, moreover, ferrous salts appear to be more stable in this solvent than in acetone. The modified method is sufficiently sensitive to measure the peroxide value of the fat of freshly drawn milk and has been applied to the study of the first stages of oxidation in milk, butter and whole-milk powder.

1. During 1950–53 the milk of 814 Ayrshire cows was sampled six times during the lactation at intervals of approximately 5 weeks, starting towards the end of the first month of lactation. The weight of milk at successive evening and morning milkings was recorded and samples from each mixed in proportion to the yield. The milk was analysed for total solids, fat, S.N.F. by difference, crude protein (N×6·38), casein and lactose.

2. The weighted lactation average was calculated for each constituent of the milk of each cow and used in genetic studies (see Part II), and in assessing the effect of age on milk composition. The analyses of the individual samples were used to determine the separate effects of stage of lactation and of season on milk composition.

3. Advancing lactation caused the following changes in milk composition:

(a) Milk yield was highest 45 days after calving and then fell regularly to the end of lactation.

(b) Total solids, S.N.F., fat, crude protein and casein contents fell rapidly for 45 days, with fat and total solids continuing to fall for a further 30 days. The concentrations of all these constituents then increased continuously for the remainder of the lactation, rising more rapidly after about 200 days.

(c) The changes in lactose content were opposite from those of fat and protein and smaller in magnitude. The changes bore a marked resemblance to those for yield. The value rose to a maximum after 45 days, fell slowly until about 165 days after calving and then more quickly. The lactose content of milk from cows in their first lactation fell much more slowly with advancing lactation than that in the milk of older cows.

4. Seasonal effects were of smaller magnitude than those arising from advancing lactation and caused the following changes in milk composition:

(a) Yield rose steadily from January to the May-June period, and then fell to a minimum during October-November.

(b) Fat content was at a maximum in October, falling steadily to a minimum in June.

(c) Crude protein and casein contents rose to a peak in May-June and again in September. Lowest values occurred from January to March.

(d) Lactose content was at a steady high level from January to June, falling to a lower level by August at which it remained for the rest of the year. Again there was some similarity in the pattern of change in lactose content and in yield. The range of variation in the values for lactose was less than those for protein and fat, and the variations were mainly opposite in sign.

(e) Total solids and S.N.F. were at a minimum in March and April, but whereas S.N.F. reached their highest level in May-June, total solids were highest in October.

5. Increasing age of the cow resulted generally in increasing yields of poorer quality milk. The difference in composition between milk from the first and the grouped ninth and later lactations was fat 0·19%, S.N.F. 0·34%, crude protein 0·08%, casein 0·21% and lactose 0·25%.

6. On the evidence of cell counts, a number of the samples came from cows with some inflammation of the udder. The average casein numbers, however, did not indicate any serious incidence of mastitis, and it was concluded that although lactose contents may have been lowered slightly, the effect of disease on the average composition was small.

7. Although the figures are not closely comparable, it appears that there has been little change in the composition of the milk of Ayrshire cows in Scotland since 1921–22.

8. The number of individual cow samples deficient in fat was 2% and in S.N.F. 16% of the total. In 560 samples of farm bulk milk none were deficient in fat and only 1·6% deficient in S.N.F. May, June and July were the months with most fat-deficient samples, and the period from January to April provided the largest number of samples deficient in S.N.F., closely followed by the period July to October.

9. Possible causes of the seasonal changes in milk composition are discussed. These changes are thought to be caused mainly by feeding although this does not give a complete explanation of all variations.

The effects of concentrations of 10–30 mM/1 of CO2 on the growth of psychrotrophic milk spoilage organisms were examined, both in untreated whole milk and in pure cultures of Pseudomonas fluorescens growing in skim-milk and broth. Spoilage of refrigerated untreated whole milk was found to be inhibited by the addition of CO2 and the effect was enhanced with increasing concentrations of CO2 and decreasing storage temperature. Experiments with pure cultures of Ps. fluorescens showed that the inhibitory effects of CO2 were not due to increased acidity or to displacement of dissolved O2, but to the presence of CO2per se which induced an increase in the duration of the lag phase of growth and had only a small effect on the logarithmic phase.

The ‘metallic compound’ previously reported as an important cause of off-flavour in oxidized dairy products has been identified as oct-l-en-3-one. The compound was found to have a flavour threshold value of one part in 109 in butterfat and one part in 1010 in water. This compound was also isolated from oxidized safflower oil.

The main requirement for milk processed in most cheese typologies is its rennet coagulation ability. Despite the increasing number of studies, the causes for abnormal coagulation of milk are not fully understood. The aim of this study was to ascertain relationships between milk characteristics and its rennet coagulation ability, focusing on the influence of calcium (Ca) and phosphorus (P). Ca and P are essential constituents of the micelles. Micellar P can be present as part of colloidal calcium phosphate (inorganic-P) or covalently bound to caseins as phosphate groups (casein-P). Eighty one herd milk samples (SCC<400 000 cell/ml) were classified as Optimal (8), Suboptimal (39) Poor (29) and Non-coagulating milk (5), according to their rennet coagulation parameters as assessed by lactodynamographic test. Samples were analysed for their chemical composition (basic composition, protein fractions, minerals and salt equilibria), physicochemical parameters (pH and titratable acidity) and rheological properties. Optimal milk was characterised by the highest contents of major constituents, protein fractions and minerals, lowest content of chloride and highest values of titratable acidity. Non-coagulating milk was characterised by the highest values of pH and the lowest of titratable acidity. At micellar level, Optimal milk showed the highest values of colloidal Ca, casein-P and colloidal Mg (g/100 g casein), while Non-coagulating milk showed the lowest values. Interestingly, there was no statistical difference regarding the content of colloidal inorganic-P (g/100 g casein) between Optimal and Non-coagulating milks. Overall, high mineralisation of the micelle (expressed as g inorganic-P/100 g casein) positively affect its rennetability. However, excessive mineralisation could lead to a reduction of the phosphate groups (g casein-P/100 g casein) available for curd formation.

Studies on the immunomodulatory properties of dietary whey proteins in mice are reported. Ingestion of bovine milk whey proteins, either as a supplement in an adequately balanced commercial diet or as the only protein source in a balanced diet, consistently enhanced secondary humoral antibody responses following systemic immunization with ovalbumin, when compared with other protein sources such as soyabean protein isolate and ovine colostral whey proteins. After 5–8 weeks of feeding, dietary milk whey proteins enhanced cell-mediated immune responses as revealed by footpad delayed-type hypersensitivity responses, and concanavalin A-induced spleen cell proliferative responses. To monitor nutritional effects of milk whey proteins, live weight, leucocyte counts and clinical changes of diet-fed mice were examined. The present results confirm other previous results that dietary bovine milk whey proteins have immunoenhancing properties in mice and these properties are unlikely to be related solely to the nutritional effects.

Aeration increased the growth and lipase production in milk by Pseudomonas fluorescens strain AFT 36, isolated from refrigerated bulk milk. A heat-stable lipase was isolated from a shaken milk culture of this microorganism by DEAE-chromatography and gel filtration in Sepharose 6B. The lipase-rich fraction from DEAE cellulose contained 3 lipases that were separated by gel filtration; only the principal lipase, which represented ∼ 71 % of total lipolytic activity, was characterized. The purified enzyme showed maximum activity on tributyrin at pH 8·0 and 35 °C; it had a Km on tributyrin of 3·65 mM. and was inhibited by concentrations of substrate > ∼ 17 mM. The enzyme was very stable over the pH range 6–9; it was relatively heat-labile in phosphate buffer in the temperature range 60–80 °C, where it was stabilized significantly by Ca2+. It was, however, very stable at 100–150 °C: the D values at 150 °C were ∼ 22 s and 28 s in phosphate buffer and synthetic milk serum respectively; the corresponding Z values in the temperature range 100–150 °C were ∼ 40 and ∼ 42 °C and the Ea for inactivation were 7·65 × 104 J mol-1 and 6·97 × 104 J mol-1 respectively.

Casein micelles of different composition were synthesized in various ways and their sub-structure was investigated with the electron microscope by means of thin sections. Earlier studies of Schmidt et al. (1974) using the freeze-fracturing technique had shown no differences in the sub-structure of natural micelles and artificial micelles containing Ca and casein only. Contrary to these results our present studies showed that for the production of synthetic casein micelles with the same sub-structure as the natural ones it is necessary to add at least 2 other ions to the casein solution besides Ca2+: these are phosphate and citrate. The citrate ions play an important role in forming the material of the dark framework of the micelles visible in electron micrographs of unstained thin sections. This supports the hypothesis of Pyne & McGann (1960) that casein micelles contain a citrate apatite.

The number of starter bacteria in Cheddar cheese was increased approximately 2 or 8 times by supplementing the normal starter inoculum with starter-cell suspensions which had been incubated with lysozyme in the absence of salt. Lysozyme-treated cells were also introduced into chemically acidified cheese in an attempt to achieve ripening in the absence of a normal starter culture. The added starters did not interfere with normal cheese-making by producing acid. The lysozyme-treated starter cells were lysed when the curd was salted and lysis was detected by the release of cell-free DNA and an intracellular marker enzyme (dipeptidase) into the cheese matrix. Free amino acid concentrations in maturing cheeses were increased up to 3 times compared with control cheeses. The intensity of Cheddar flavour was not increased in starter cheeses by the presence of additional lysozyme-treated starter and no Cheddar flavour developed in chemically acidified cheese containing the lysozyme-treated cells. It is concluded that intracellular starter enzymes play no direct part in flavour formation, but produce breakdown products from which Cheddar flavour compounds may be formed by other unknown mechanisms.

Raw whole milk of high microbial quality ([les ]4×104 cfu/ml) was processed using a ~15 l/h homogeniser with a high pressure (HP) valve immediately followed by cooling heat exchangers. The effects of homogenisation between 100 and 300 MPa (HP valve) with an initial milk temperature Tin=4 °C or 24 °C was investigated on the inactivation of: (i) endogenous alkaline phosphatase (ALP); (ii) endogenous milk flora and (iii) two Gram positive (Listeria innocua and Micrococcus luteus) and one Gram negative (Pseudomonas fluorescens) strains inoculated into milk. Temperatures T1 and T2 measured before and immediately after the HP valve, and fat globule size distributions were also determined. ALP activity slightly decreased after homogenisation above 250 MPa when Tin=4 °C (corresponding T2>58 °C), but markedly decreased above 200 MPa when Tin=24 °C (T2>60 °C). In contrast to inactivation induced by continuous short-time thermal treatments, ALP inactivation induced by HP homogenisation was clearly due to mechanical forces (shear, cavitation and/or impact) in the HP valve and not to the short ([Lt ]1 s) residence time at temperature T2 in the same valve. Inactivation of the three exogenous microorganisms led to similar conclusions. Homogenisation at 250 MPa or 300 MPa (Tin=24 °C) induced a 2–3 log cycle reduction of the total endogenous milk flora and a 1·5–1·8 log cycle reduction of inoculated List. innocua. Higher reduction ratios (2–4 log cycles) were obtained for the two other microorganisms. The highest levels of ALP inactivation corresponded to the highest extents of microbial reduction. Running the milk twice or three times through the homogeniser (recycling), keeping temperature T1≈29 °C and pressure=200 MPa, increased homogenisation efficiency.

The method of assaying rennin described by Berridge(1) requires the formation under reproducible conditions of a thin flowing film of milk. Although the skill needed to keep the film flowing at the optimum rate as the clotting point is approached is readily acquired, an automatic method has the advantage of complete reproducibility. A suitable film can be maintained if 10 ml. of milk are placed in a boiling tube of 2·2 cm. internal diameter and the tube, with its axis inclined at about 30° to the horizontal, rotated at 3·0 r.p.m. In the apparatus now used, the boiling tube is mounted on a rubber bung, which in turn is mounted concentrically on a spindle connected through Meccano gears to a 'Sangamo' electric clock motor. A special mechanical coupling allows the bung to be rotated by hand one revolution forwards, thus allowing 20 sec. for fitting or releasing the boiling tube before the drive is again taken up.

Milk deposits from an ultra-high-temperature (UHT) plant were analysed for protein, fat and minerals. The physical appearance and composition of the deposits was found to vary with modifications of the plant. For each particular arrangement of the plant, a characteristic pattern of deposits occurred. Possible mechanisms involved in the formation of the deposits are discussed.