Five hundred and seven samples of iso-electric caseins were typed by starch-gel electrophoresis and their polymorphic compositions were determined in terms of αs1-, β- and κ-casein variants. Clotting times in minutes were determined on 50 ml of 3% casein solution in milk dialysate containing 0·02 M-Ca as CaCl2 and 2 mg commercial rennin powder at pH 6·60–6·80. The clotting times ranged from 4·06 to 7·41 min. The data indicated that the polymorphic composition of casein significantly influenced its stability to rennin. The polymorphic combinations αs1B+βA+κB and αs1B+βAC+κAB, were the most resistant to rennin action (clotting times of 7·41, 7·39 min respectively). The combinations αs1B+βB+κAB and αs1BC+βB+κB were the least resistant to rennin action (clotting times of 4·25 and 4·06 min respectively). In most cases, caseins with lower stability to rennin contained κB or βB or both in their composition.

Two chemical tests were used to measure the extent of lactose degradation by Maillard browning in casein and skim-milk powder during storage. At the same time these products were evaluated organoleptically. Highly significant correlations between age, flavour score and level of browning in skim-milk powder were in contrast to the lack of any relationship between the flavour score or age of casein and its level of browning. Maillard browning occurring during storage of skim-milk powder was accompanied by increasing levels of cereal and malty flavours. A stale flavour, which closely resembled the musty flavour common to casein, was not observed in skim-milk powder until after storage for 9 months. A distinct difference in the flavour stability of rennet casein and acid casein could not be accounted for in terms of the relative extent of lactose degradation. Evaluation of casein samples with different lactose contents suggested that Maillard browning does not contribute to flavour defects during storage unless the curd is so insufficiently washed during manufacture as to produce an abnormally high level of lactose in the dried product.

Two possible hypotheses for the mechanism of milk clotting were tested. The results obtained constituted strong evidence against one and suggested that the second is improbable. Milk was separated into a 5-fold-concentrated casein micelle suspension and milk serum. Pre-renneting of the serum did not reduce the rate of clotting on subsequent addition to the micelle suspension whether or not the conditions were such that the para-κ-casein became extensively aggregated. Washing of casein micelles up to 3 times with milk dialysate at 23°C extracted very little casein from the micelles and did not increase the clotting time of micelles resuspended to about the same concentration as in milk. The results appear to constitute decisive evidence against the hypothesis of milk clotting proposed by Parry & Carroll (1969). S-carboxymethyl-κ-casein, S-carboxymethyl-κ-casein containing 2·5 dimethylaminonaphthalene sulphonyl residues per mole, and rennin-treated dimethylaminonaphthalene sulphonated-S-carboxymethyl-κ-casein all bound Ca to the same extent at 30°C and pH 6·5, over the range 0·5–15·3 mM-CaCl2. This adds support to existing evidence that milk clotting does not involve formation of Ca bridges between casein micelles.

The variations in the composition of milk over an 18-week period from lactating British Alpine and Anglo–Nubian goats imported into Trinidad are reported. The animals were fed Napier grass (Pennisetum purpureum) ad lib. and 2 lb of a proprietary concentrate mixture daily.

The mean percentage contents of the milks of British Alpine and Anglo–Nubian goats were: total solids 11·49±0·22, 12·17±0·28; butterfat 3·42±0·18, 4·06±0·22; total N 0·462±0·04, 0·530±0·07; ash 0·78±0·02, 0·79±0·03 and ‘lactose’ 4·38±0·28, 4·05±0·55. Anglo–Nubian goats had a higher content of percentage total solids, fat, total N and ash. For all constituents the coefficient of variation between weeks was higher for Anglo–Nubian goats. No significant trends were found between weeks for each of the components analysed for both breeds. Statistically significant correlations (P < 0·01) were found between percentage fat and percentage total N for both breeds.

The mean mineral contents (mg/100ml) for the milks of British Alpine and Anglo–Nubian goats were: Na 52·6±4·6, 55·5±1·9; K164·0±8·4, 133·4±11·4; Ca 85·5±11·6, 89·9±9·8; Mg 13·2±3·8, 10·2±1·6 and P 74·7±3·9, 82·1±3·9. British Alpine goat's milk had a higher content of K and that of Anglo–Nubian goats had a higher content of P; both differences were statistically significant (P < 0·01).

The milk composition figures in this study together with those from a similar study in Guadeloupe suggest that the values are low and considerably lower in comparison with the same breeds of goats in a temperate environment. This could be a feature of dairy goats imported into the tropics due perhaps to inadequate nutrition. Much more information is needed to verify this possibility.

An isothermal jacket calorimeter was set up and apparent heat capacities of skim-milk, half-and-half milk, whole milk, and their concentrates up to about 30 % total solids were measured throughout the 5–70°C temperature range. Above the butterfat melting interval, the heat capacity of every sample was linearly related to temperature and a linear regression equation was found to relate heat capacity and total solids content for each one of the 3 milk sets. By means of an overall approximation, a single expression relating heat capacity, temperature and total solids content was also derived. To facilitate calculations of relative heat content for practical purposes, an average apparent heat capacity was computed for the whole temperature range and a nomogram was constructed for its determination from fat and solids-not-fat contents in any sample.

The effect of heat treatment on the rennet coagulation times (RCTs) of various casein–whey protein systems, the effect of pre-renneting centrifugal serum on the RCT of casein subsequently mixed with such serum, the influence of repeated centrifugation and resuspension at constant protein level on the RCT of such suspensions, and the rate and completeness of the hydrolysis of sialic acid in micellar and non-micellar caseins, all lead to the conclusion that κ-casein does not occur in milk in 2 functionally different states, as proposed by Parry & Carroll (1969).

A lactating mammary gland of a goat was perfused for 9 h in the presence of [U-14C]-l-threonine and received adequate quantities of glucose, acetate and amino acids. Two lactating sheep udders were likewise perfused in the presence of [U-14C]-l-phenylalanine: the plasma levels of phenylalanine in the first of these experiments were 4 times higher than in the second.

In the [14C]threonine experiment, 4 % of the casein and 0·4 % of the expired CO2 were derived from threonine; 85 % of the threonine and 1·6 % of the glycine residues in casein originated from plasma threonine. Small 14C levels were found in glutamic acid, aspartic acid and serine of casein. The relative specific activities amongst the casein amino acids and the appearance of appreciable labelling in plasma glycine are consistent with the view that threonine is split by threonine aldolase.

In the [14C]phenylalanine experiments virtually no radioactivity was detected in CO2, lactose or citric acid, indicating that this substrate is not broken down by mammary tissue. In the second experiment, 96 % of the phenylalanine and 0·3 % of the tyrosine of casein originated from plasma phenylalanine. In the first experiment, a 30-fold higher 14C incorporation into casein tyrosine relative to phenylalanine was observed. The possible significance of the phenylalanine concentration in the plasma on the degree of conversion of phenylalanine into tyrosine within the mammary gland is discussed.

The total solids (TS) content of milk was estimated using freeze-drying followed by a Karl Fischer (KF) titration. The average solids-not-fat (SNF) value for 12 milks calculated from the TS results by this method was 0·003±0·039 % lower than that calculated from the TS results by the British Standard (1963) gravimetric method (BS). The average SNF value calculated from the sum of the contents of lactose anhydride, ‘total’ protein and ash was slightly higher (+0·017±0·068 %) than that obtained from the BS method.

Certain modifications of the BS method for TS gave erroneous results. Heating 1 or 2 g milk at an oven temperature of 100 °C for 5 h, however, gave results agreeing fairly closely with the BS values.

For lactic (starter) casein whey the average SNF content from freeze-drying and KF titration was 0·075 % higher than that obtained by following the BS procedure (for TS in milk). Average values for the sum of the SNF constituents (including lactic acid) were 0·08–0·11 % lower than those obtained from BS method.

In many of the values recorded in the literature there is an unexplained discrepancy between the SNF content calculated from the sum of the constituents and that calculated from the TS content estimated gravimetrically by an alternative British Standard (1951) method. The need for a uniform method for estimating TS is stressed.

Ten different types of milk, arranged in 3 sets, were observed to be pseudoplastic fluids with linear behaviour at high shear rates in a coaxial cylinder rheometer; at low rates of shear all the samples could be considered sufficiently Newtonian and a Höppler viscometer was used for their study. Dynamic viscosity coefficients of skimmilk, half-and-half milk, whole milk and their concentrates up to about 30% total solids were determined every 5°C between 0 and 80°C. For every milk type a quadratic equation relating t to log η was obtained. At a given temperature the concentration dependence of η for the 3 milk sets was expressed by a single relation of the same mathematical model as in the previous case. A general expression was derived to enable η to be calculated at any temperature (0–80°C), and for any concentration (0–30% total solids) whatever the fat to solids-not-fat ratio (0·01–0·4). A nomogram was also constructed for the immediate computation of η without any practical loss of accuracy.

Storage of aseptically packed ultra-high-temperature (UHT) milk has been shown to be accompanied by pronounced changes in the molecular weight distribution of the pH 4·6 insoluble casein. These changes were both time-dependent and temperature-dependent, proceeding much more rapidly at 37 °C than at 30 °C, but even at 4 °C the changes were considerable. The proportions of high molecular weight material present before and after storage have been studied by gel filtration in dissociating solvents using Sephadex G-200. Measurements have also been made on casein material isolated from in-bottle sterilized milks and canned evaporated milk for comparison. The results of gel filtration have been compared with sedimentation coefficients recorded by ultracentrifugation and with changes in the content of amino groups titratable with trinitrobenzene-sulphonic acid. The results are compatible with the suggestion that the Maillard reaction occurred at ambient temperatures, and over a period of several months led to browning and sediment formation due to covalent cross-linking of polypeptide chains. The implications of this are discussed with reference to spoilage of UHT milk on storage and the phenomenon of gelation.

A method is described for determining alkaline phosphatase activity in cheese using phenolphthalein monophosphate as substrate. Results are read with a Lovibond comparator after 1 h incubation. Advisory standards are proposed based on a minimum pasteurization for 15 s at 71°C of the milk used for cheese-making.

Three enzymes were assessed as rennet substitutes for cheese-making. The bovine and chicken pepsins used were relatively crude extracts of bovine stomach mucosa and chicken proventriculae respectively; the swine pepsin was a partially purified commercial product. The ratios of milk-clotting activity to general proteolytic activity were high for rennet and bovine pepsin and low for swine and chicken pepsins. Both bovine mucosa and chicken stomach gave low milk-clotting activities compared with calf stomach. For all the enzymes the chemical reactions causing milk clotting appeared to be the same. The milk-clotting activity showed a decrease with increase in substrate pH for all the enzymes, although they were all still active at pH 6·81.

Duplicate cheeses were made from each of the swine, bovine and chicken pepsins, with rennet as a standard in each trial. The cheese-making process was similar with each enzyme, but differences appeared during ripening. The chicken-pepsin cheeses had poor body and weak Cheddar-cheese flavour, with many and intense off-flavours. The cheeses made with bovine and swine pepsins were only slightly inferior in quality and intensity of Cheddar-cheese flavour to the rennet cheeses. From a simulated cheese-making experiment it was concluded that 30–40 % of the added rennet, bovine pepsin and chicken pepsin was probably inactivated during the cheese-making process and that most or all of the swine pepsin was lost. These results provide an explanation for the variations observed in cheese ripening.

It was concluded that chicken pepsin would not prove a suitable rennet substitute for making Cheddar cheese because of the quality of the cheese produced, and that bovine pepsin would not prove suitable because of the cost of preparing a suitable extract. Swine pepsin would appear to be suitable if the ripening time were to be lengthened or if another enzyme were to be added to assist ripening; it is cheaper than rennet and other rennet substitutes.

Radioactive labelling of intact fat globules with 125I gave at least one labelled protein in each of the 4 fractions of the fat globule membrane. Similar treatment of isolated membrane gave extensive labelling of all membrane proteins in each fraction.

It was concluded that all fractions are partially exposed at the membrane surface and the existence of well-defined layers is unlikely.

The effect of temperature and time of heating whole milk on the renninclotting time, the primary phase of rennin action and the protein (mainly β-lactoglobulin) soluble in 2% trichloroacetic acid (TCA), have been studied. Considerable changes in these parameters occurred above 60°C. The primary phase was inhibited (the degree of inhibition being both temperature and time-dependent), the clotting time was increased, and the protein soluble in 2% TCA decreased considerably.

It is suggested that the inhibition of the primary phase was due to complex formation between κ-casein and β-lactoglobulin, the increase in clotting time to a combination of complex formation and a change in the distribution of Ca, and the decrease in β-lactoglobulin to both its interaction with κ-casein and its heat denaturation. The relevance of such changes to the heat stability of milk is discussed.

Details are given of a spectrophotometric method for determining nitrite in cheese and other dairy products at levels down to 0·05 μg NO2-N/g and of a spectrofluorimetric method for levels down to 0·003 μg NO2-N/g. Nitrite-N levels in cheese varied from 0·003 μg/g to 0·021 μg/g. These levels increased up to 4-fold after UV irradiation of the cheese extracts. No nitrite (< 0·003 μg/ml) could be detected in raw milk.

The temperature coefficient of electrolytic conductivity of normal milk decreased with temperature from 0·0241 ± 0·0003/°C at 15 °C to 0·0173 ±0·0003/°C at 40 °C. The temperature coefficient for a sample of mastitic milk lay within this range.

The effect of ageing fresh milk for 24 h at 4°C on the composition of 4 fractions of milk fat globule membrane (FGM) – microsomal pellet membrane (MPM), deoxycholate soluble membrane (DOCM), high density membrane (HDM) and low density membrane (LDM) – prepared from washed cream treated with sodium deoxycholate (DOC), was studied for milk of individual cows. Total FGM and its fractions were solubilized by treatment with sodium dodecyl sulphate (SDS), EDTA and β-mercaptoethanol, and the dissociated FGM proteins were separated by polyacrylamide gel electrophoresis in the presence of SDS.

Ageing resulted in a greater loss of phospholipid during cream preparation than was found with fresh milk, and also in a reduction in the total amount of FGM isolated. Loss of FGM material was confined to MPM, DOCM and LDM and was accompanied by compositional changes in DOCM and LDM, quantitatively the most significant fractions. Ageing also produced changes in the gel electrophoresis patterns of DOCM, where the band of greatest mobility (mol. wt about 16000) was partially lost, and of LDM, where there was an increase observed in the major protein band.

The implication of the results on the proposed models of FGM structure is discussed.

Twenty-four Murrah buffalo cows were divided into 4 groups and were fed 0·3, 0·4, 0·5 or 0·6 kg of concentrate mixture per litre of milk produced. Energy and N balance studies were conducted on 2 animals from each group in the sixteenth and seventeenth weeks of the study. The efficiency of conversion of gross energy into milk energy ranged from 10·9 to 12·7%. The efficiency of conversion of metabolizable energy (ME) and ME above maintenance into milk energy ranged from 25·0 to 32·2% and from 51·7 to 78·6% respectively. Differences amongst groups were not significant for all the 3 parameters.

The conversion of digestible N into milk N varied between 23·6 and 27·3% in different groups. The efficiency of utilization of metabolizable N for milk production ranged between 47·6 and 56·4%. Various groups however did not differ significantly from each other in these respects.