A new bioassay for the detection of penicillin in milk which can be completed in 1½ h has been developed. The test depends on measurement of pH changes in milk produced by an antibiotic sensitive organism, Streptococcus thermophilus BC. To measure the changes potentiometrically would require a very sensitive pH meter. Change in the enzymic coagulation time, which is extremely dependent on pH, was used in the present study to measure small changes in pH. The test is highly reliable and should be suitable as a milk selection test.

The susceptibility of the casein in milk to proteolysis was shown to be greatly influenced by its state of aggregation. In normal milk, where the casein is largely in micellar form, the αs1- and β-caseins are almost inaccessible to proteolysis. On removal of the colloidal phosphate, the casein micelles disintegrate, rendering the components, especially the αs1-casein, accessible to proteolysis. The role of colloidal calcium phosphate in the casein micelle is believed to be that of a non-specific aggregating agent which can be effectively replaced by calcium. Dissolved colloidal phosphate can be effectively reformed by elevation of the pH of colloidal phosphate-free (CPF) milk before equilibrium dialysis. Addition of κ-casein to CPF milk also causes aggregation of the component caseins but the micelles formed are smaller than those of normal milk.

The behaviour of micellar β-casein differs considerably from that of micellar αs1-casein. The evidence suggests that part of the β-casein freely dissociates either outside or within the micelle when the temperature is reduced. The temperature dependence of the susceptibility of β-casein to proteolysis was similar in skim-milk and in solutions of sodium caseinate, and increased as the temperature was reduced. αs1-Casein was quite resistant to proteolysis in normal milk but became susceptible when the micelle structure was disrupted on removal of colloidal phosphate.

It is concluded that limited proteolysis may prove a valuable technique in the study of casein micelle structure.

A study was made of the changes in the iodine value of milk fat from cows following the feeding of linseed oil (450g) at intervals of 6 days. Three cows in midlactation and of differing age and breed were used. The experiment consisted of a series of 3 collection periods, each of 6 days' duration, with 12 days between periods. Three doses of oil were given (6 days apart) between periods 1 and 2, the last dose of oil being given immediately before the start of period 2. The mean iodine values for the 3 periods were 35·5, 40·5 and 34·7 respectively. There was a distinct pattern of change in iodine value with time after feeding the oil, a maximum of 45·5 being attained on the second day with a subsequent decline to 38·5 on the sixth day. Analysis of variance showed that these effects were significant and due to oil-feeding, whereas changes in yield and content of fat and production of milk could be accounted for by normal decline in lactation. It was concluded that the periodic feeding of a large dose of linseed oil produces a significant increase in the iodine value of milk fat and avoids the possible disadvantages of continuous fat supplementation.

The modification of arginine residues in casein by treatment with glyoxal at pH 8·6 resulted in an inhibition of the coagulation of rennin-treated casein. The effective residues are on the κ-casein fraction and inhibition of coagulation was complete when about 1·5 residues of arginine/mole of κ-casein had been altered. It is suggested that the arginine forms part of a positively charged region that also includes lysine and histidine residues, and that this region is important in the coagulation of the casein.

The effects of intravenous infusion of insulin and of sodium succinate on milk yield and composition and on the yields of milk constituents was investigated in the goat. Depression of plasma glucose concentration to less than 40 mg/100 ml in response to infusion of insulin was associated with a decrease in milk volume and lactose yield, and increases in the contents of fat and casein and in the yield of fat. The increase in fat yield was the result of an increased output of C12–C18 fatty acids; the output of C4–C10 acids was slightly reduced. Infusion of succinate caused a reduction in the yields of milk and of lactose, protein and fat. There was no consistent change in milk composition.

The origin of the effects is discussed.

A method is described for estimating the growth of Group N streptococci in milk and other media by measuring the 260 nm-absorbing material extracted from the cells with 0·1 M-NaOH at 100°C. A good correlation between the absorbance at 260 nm of the hot alkali extracts and the dry weight of the cells was obtained for cultures of Streptococcus lactis during the logarithmic and early stationary phases of growth.

Lactose in the plasma of 12 lactating goats and one cow with healthy udders has been measured by a sensitive and specific enzymic assay. Mean values (±S.E.) for arterial and venous concentrations were 52±11 µM and 70±11 µM respectively. Simultaneous measurement of mammary blood flow, milk yield and milk lactose concentration showed that 2·70±0·25% (mean±S.E.) of the total lactose production passed into the plasma. This rose to 42·7% in a goat when milking was suspended for 46 h. No lactose was found in the plasma of male and non-lactating female goats, but it was detected 4 days before parturition in a nulliparous goat.

An experimental ultra-high-temperature milk sterilizer, of 1140 1/h capacity and capable of operation as a plate-type indirect heating plant or as a steaminto-milk direct heating plant, has been installed for comparisons of the milk produced by the direct and indirect processes. The sterilizer and its ancillary equipment are described, together with methods of plant sterilization and milk processing. Time-temperature profiles of the plant are given for both modes of operation.

The variation of sporicidal efficiency with processing temperature was determined for an experimental ultra-high-temperature (UHT) milk sterilizer operating alternatively as an indirect or as a direct heater. Whole milk was inoculated with large numbers of spores of Bacillus subtilis 786 and Bacillus stearothermophilus TH24, and the proportion of spores surviving the sterilizing process was calculated from dilution and colony counts on the untreated and treated milk. The results for B. subtilis spores were unreliable, and the dilution count results for B. stearothermophilus spores were influenced by the inhibitory effect of the UHT processed milk. The results for the colony counts of B. stearothermophilus spores were preferred as a basis for the comparison of the direct and indirect processes. Over the range of processing temperatures 137–145 °C it was found that the sterilizing temperature had to be 3–4 degC higher with direct heating than with indirect heating to give equal spore destructions.

The bacteriological results of Franklin, Underwood, Perkin & Burton (1970) are analysed to show that colony counts of Bacillus stearothermophilus spores in UHT-treated milk are influenced by the inhibitory action of the milk, so that the sporicidal effects of the UHT process as calculated from the results are too high. Only the elimination of the inhibitory factor will allow true sporicidal effects to be determined.

The rates of denaturation by heat of α-lactalbumin and β-lactoglobulin in skim-milk were measured by an immunodiffusion method over a wide range of temperatures. Both reactions showed an unusual temperature dependence.

The denaturation of α-lactalbumin is a first-order reaction; between 90 and 155 °C the kinetic constant k1 in s−1 is given by the equation

where T is the temperature in °K.

The denaturation of β-actoglobulin in skim-milk is second order with respect to time, and the kinetic constant k2 in 1g–1 s–1 is given by 2 equations, valid for different temperature ranges. Between 68 and 90 °C,

Between 90 and 135 °C

Results obtained by the immunodiffusion method agreed well with those found by salt fractionation of the milk proteins. The denaturation rates decreased when a specific reagent for sulphydryl groups was added, suggesting that such groups are involved in the denaturation of both proteins.

Study of the dissociation of high-molecular-weight aggregates of preparations of αs1-, β-, κ-, and para-κ-casein by the detergent, sodium dodecyl sulphate (SDS), showed that there are differences in the aggregation properties of the individual caseins. Binding of detergent led first to the dissociation of casein aggregates and then to further interaction with the casein molecules. The amounts of detergent required to give the minimum sized protein-detergent aggregate when expressed as mg/mg casein were similar for κ-, para-κ- and αs1-casein but much less for β-casein. However, expressed as mole/mole the requirement for κ- and αs1-casein was similar but was twice that found for para-κ- and β-casein. The maximum amount of SDS bound was about twice that required for complete dissociation of the aggregates for κ-, para-κ- and αs1-casein but was 13 times greater for β-casein.

Complete dissociation of κ-casein aggregates by SDS alone was not possible due to the presence of aggregates formed by disulphide linkages. These aggregates, which consisted of 3±1 protein molecules, accounted for about one-third of the κ-casein in the preparations examined.

The dissociation of casein aggregates by the detergent sodium dodecyl sulphate (SDS) gave rise to difference spectra and these spectra were characteristic for each of the different types of casein. Increase in absorption by the chromophore groups, tyrosine and tryptophan, when αs1- and β-casein aggregates were dissociated indicated binding of the detergent at regions of the molecule containing these residues. A decrease in absorption when κ-casein was dissociated indicated that the tyrosine and tryptophan residues were not in the region of the molecule to which the detergent was bound and that in the κ-casein aggregate these residues were in a more hydrophobic environment. Peaks on the difference spectra were obtained at 280 and 288 nm for αs1-casein and 284 and 291 nm for β-casein and troughs at 278 and 286 nm for κ-casein. The difference spectrum reached a maximum value when the αsl- and β-casein aggregates were dissociated and the further binding of SDS did not alter this value. The large negative change in the difference spectrum of κ-casein did not occur until after most of the aggregates were dissociated and did not reach a maximum until binding with SDS was complete. The value obtained for ΔOD was found to be temperature-dependent for β-casein-SDS interaction, but not for αs1- and κ-casein. Changes in spectra were also observed when αs1- and κ-casein interacted to form aggregates. The data obtained confirmed the importance of hydrophobic binding in casein aggregate formation and indicated the possible involvement of tyrosine and tryptophan residues in this binding.

Carboxypeptidase A (CPA, mol. wt = 34600) and 1-dimethylaminonaphthalene-5-sulphonyl chloride (dansyl chloride, mol. wt = 270) were shown to react with the 3 casein subunits, αs1-, β- and κ-caseins, which constitute the micelle. Virtually all of the C-terminal amino acids were released by CPA from these subunits without disrupting the micellar structure.

It was verified that the rate of dissociation of the micelle into low molecular weight complexes is considerably slower than the rates at which these reagents act on micellar components. It is concluded that dansyl chloride, CPA, and probably also rennin, whose molecular weight is similar to that of CPA, are able to penetrate to the interior of the micelles. This was confirmed in a quite independent way with a non-reacting agent, myoglobin (mol. wt = 17000).

It is postulated that the casein micelle has a sponge-like structure in which the 3 different subunits are distributed fairly uniformly.

Micellar fractions obtained by differential centrifugation from skim-milk and colostrum were examined for their content of αs1-, β-, κ- and para-κ-caseins by the use of CPA as described by Ribadeau Dumas (1968). The results confirm that there is a real difference in κ-casein content of the micelles, according to their rate of sedimentation.

The distribution and activity of alkaline phosphatase (E.C. 3.1.3.1), acid phosphatase (E.C. 3.1.3.2), catalase (E.C. 1.11.1.6), xanthine oxidase (E.C. 1.2.3.2), aldolase (E.C. 4.1.2.7 and 4.1.2.13), ribonuclease (E.C. 2.7.7.16) and carbonic anhydrase (E.C. 4.2.1.1) were studied in the major components of bovine milk. Fractionation was accomplished by centrifugation of milk, skim-milk and buttermilk, and ammonium sulphate precipitation of skim-milk serum. The range of activities found for the enzymes studied are tabulated together with the activities of some of the enzymes in mastitic milk, and the significance of the results obtained is discussed. No carbonic anhydrase activity was detected in any of the samples tested. The other enzymes studied were found to have a greater proportion of their total activity located in the skim-milk fraction. However, all of these enzymes except ribonuclease had a higher specific activity in the fat fraction.

A description is given of a simple milking apparatus for collecting the yields of individual ewes, and of the milking procedure used in studies of lactation in ewes of ‘mutton’ breeds. The incidence of mastitis, level of milk yield, some of the non-nutritional factors affecting the milk yield and the coefficients of variation of milk yield and composition are discussed in relation to data from 6 experiments in which ewes of mutton breeds were machine-milked twice daily for 6–9 weeks of lactation.

The composition of ewe's milk during the first 4 days of lactation and on the 100th day of lactation was investigated. The total fat content was highest (17· 9%) on the day of parturition but decreased rapidly to reach a level on the 2nd day after parturition that was similar to that observed on the 100th day of lactation (9·9 %).

The concentration of octadecenoic acid, which was the major fatty acid of ewe's milk, was very much higher in early lactation than in late lactation. As the concentration of octadecenoic acid decreased the concentration of the shorter chain fatty acids (6:0−14:0) increased. The major octadecenoic acid was the cis-9 isomer. However, the proportion of the trans-11 isomer increased from 5·5 % of the total octadecenoic acid concentration in early lactation to 11·9 % in late lactation. Although linoleic acid remained a minor component of the fatty acids of the milk during lactation, its concentration increased from less than 1 % during early lactation to 1·4 % by the 100th day of lactation.