Cows were fed either 75 or 100% of the recommended intake levels for protein and 100% of recommended energy levels (Agricultural Research Council, 1965) from 8 weeks pre-calving until 14 weeks post calving. From 14 weeks post calving and to the end of lactation all the cows received 100% of the recommended protein and energy intakes.

The mean of the 305-d milk yields of the 2 groups was not significantly different and although cows on the lower protein intake produced less lactose during the first 14 weeks of lactation there was no significant difference in total lactose, fat, protein or total solids production between the groups. In both groups blood packed-cell volume, red cell count and haemoglobin decreased during the first 10 weeks of lactation and then began to increase in the high-protein group. The cows receiving the low-protein diet showed a similar increase only when they received the high-protein ration from 14 weeks post calving. The mean interval from calving to conception was 27·5 weeks in the high-protein group and 20 weeks in the low-protein group.

It is concluded that feeding 75% of protein requirements to dairy cows during the first 14 weeks of lactation does not reduce milk yield or quality significantly and probably has no adverse effect on fertility.

This paper is a sequel to a previous one in which a number of factors likely to influence the accuracy of counting somatic cells in milk was assessed; in the present work the effects of 5 other factors are investigated. In a study of storage time and temperature of milk samples fixed in formalin, a significant increase in cell count occurred after 5–7 d when samples were stored at room temperature (17–23°C), compared with those maintained at 4 °C. When manual and mechanical mixing of fixed samples were compared only marginal differences in cell counts were observed. An increase in cell counts followed manual dilution of milk samples in comparison with automatic dilution. The temperature of samples prepared for counting was also studied and no significant variations occurred between mean temperatures of 12·7 and 32·9 °C. The final factor evaluated was that of holding time before counting; using 4 cell-count levels it was observed that counts were acceptable up to 1½ h.

Chemical analyses, polyacrylamide-gel electrophoresis and isoelectric focusing of milk-fat globule membrane proteins (FGMP) obtained from the milk of 2 Murrah buffaloes were done to determine if any change in composition occurred during lactation. Changes in the levels of sialic acid, hexose, hexosamine, N and P were found in the FGMP obtained at different stages of lactation. On the day of parturition, 8 major proteins in FGMP were determined by sodium dodecyl sulphate polyacrylamide-gel electrophoresis whereas 6 major proteins were obtained in FGMP of middle and late lactation milks. Isoelectric focusing of FGMP showed 8–9, 9–13 and 13–16 proteins from colostrum, middle and late lactation milks, respectively and the isoelectric pH of the proteins varied from 5·25 to 7·80, 5·85 to 8·30 and 5·75 to 8·61 respectively.

Properties of whole milk and milk fractions from cows fed a diet that gave a greatly increased proportion of unsaturated fatty acid residues (especially of linoleic acid) in the milk lipids were studied, and this milk (high-linoleic milk) was compared with milk from cows on a control diet (control milk). The milk fractions were isolated by high-speed centrifugation of whole milk or cream and were examined by chemical analysis and electron microscopy. During centrifugation the globules of milk fat were disrupted and the membranes (fat-globule ‘ghosts’) floated as a layer beneath the free lipid. Membrane proteins from the 2 sorts of milk gave the same electrophoretic pattern and the amino acid compositions were the same. Lipid analysis of the membrane fraction from high-linoleic milk showed the expected increase in the proportion of unsaturated fatty acid residues in the neutral lipids, but there was an unexpected decrease in the proportion of unsaturated residues in the membrane phospholipids. No differences were found between high-linoleic and control milk in the ultrastructure of the milk-fat globules or the isolated membranes.

Milk-fat globule membrane (MFGM) proteins were solubilized by treatment with SDS. Four of the major proteins were isolated as SDS complexes using column chromatography. The purity of each isolate was determined by SDS polyacrylamide gel electrophoresis and sufficient of each protein was obtained for amino acid analysis. The amino acid compositions of the isolated MFGM proteins and a total MFGM protein extract were determined. Differences in amino acid composition were found in particular between the major MFGM glycoprotein and the other 3 membrane proteins. The relationships of the amino acid composition to protein properties and structure are discussed.

The effects of heat and pH on sulphydryl (–SH) and disulphide (–SS–) groups of β-lactoglobulin (β-lg) A have been studied by heating at different temperatures and pH values in air and at pH 6·9 in the absence of air. At pH 6·9 under aerobic conditions a decrease of –SH groups and an increase of –SS– groups was observed with increasing time and temperature. Only small changes were found under anaerobic conditions. At pH from 3·0 to 9·8 the –SH level decreased while the –SS– level increased up to pH 6·9 and then dropped rapidly. In addition to –SH/–SS– interchange there were reactions to other sulphur-containing compounds as seen from the losses in the total amount of –SH plus –SS– sulphur. The results of gelchromatographic investigations suggest that –SH-initiated –SS– exchange-reactions play an important role in the formation of high molecular weight polymers of β-lg A during heat treatment, and that the formation of intermediates depends on the presence of air.

The heat denaturation of α-lactalbumin (α-la) in NaCl and KCl solutions, milk ultrafiltrate and milk was studied using the method of micro complement fixation. It was established that this protein was very resistant to heat denaturation and that it was more stable in milk ultrafiltrate than in the other media studied at temperatures up to 70 °C. Of the various milk proteins added to α-la, only β-lactoglobulin (β-lg) formed a heat-induced complex with this protein. This complex was identical in milk ultrafiltrate or in milk and depended on the molar ratio between both proteins; it was not modified by any other milk proteins. The binding of a-la to β-lg changed the ability of the latter protein to bind κ-casein.

Ovine and caprine milks showed a marked stability maximum at ~ pH7 in their heat-stability/pH curves, but became very unstable at higher pH values. A low level of κ-casein appears to be responsible for this low stability of ovine milk; β-lactoglobulin offsets the stabilizing influence of κ-casein at lower pH values. Removal of colloidal calcium phosphate from ovine or caprine milks had very little influence on their heat stabilities and it was necessary to reduce the concentration of soluble salts to a very low level before an effect was observed. At the pH of maximum stability ovine and caprine milks, although quite variable, had stabilities in the same range as bovine milks. Equine milk was very unstable to heat and the shape of the heat-stability/pH curve of most individual samples was similar to ovine and caprine milks.

β-Caseins isolated from buffalo's and cow's milk were hydrolysed either with rennet or with microbial proteases from Mucor miehei, M. pusillus Lindt or Endothia parasitica. The degradation products were separated by polyacrylamide-gel electrophoresis and the residual β-casein was determined quantitatively after various times. The electrophoretic patterns of the degradation products of buffalo and bovine β-casein produced by the different enzymes were not identical. β-Casein of buffalo's milk was hydrolysed by rennet and M. miehei protease at a slower rate than that of cow's milk. The reverse was found with E. parasitica and M. pusillus Lindt proteases. Carbamylation of buffalo β-casein was found to retard its proteolysis by all the enzymes but particularly by rennet and M. miehei protease.

Clones of Streptococcus lactis and of Str. cremoris resistant to some bacteriophages that could virtually annihilate the parent strains were selected by plating very large numbers of the bacteria in the presence of the phage. Some of the resistant strains were shown to form acid rapidly in milk and to be satisfactory for cheese-making. They were not lysogenic. Adsorption of the phage to which the parent culture had been sensitive still occurred with 3 strains but not with 4 others. Rates of mutation from phage sensitivity to phage resistance were determined for 2 original strains of Str. lactis and were found to be of the same order as the rates quoted in the literature for other mutations. The mutation rates of phages to forms which could attack the resistant strains were too low to be measured.

The highest incidence of lipolytic activity among the psychrotrophic Gram-negative flora of commercial raw milks was found in strains of Pseudomonas fluorescens and Ps. fragi. The lipases of all of the lipolytic strains remained wholly or partly active after heat treatment at 63°C for 30 min. Two of the strains tested further had lipases which retained 20–25% of their activity even when heated at 100 °C for 10 min. Cheeses made from milks in which strains or a single strain of lipolytic Gram-negative rods (GNR) had been allowed to multiply to > 107 colony forming units/ml became rancid after 4 months even though the GNR had been killed by pasteurization. The rancidity was characterized by a soapy off-flavour in cheeses containing free fatty acid concentrations from 3 to 10 times higher than those in control cheeses made from stored milks with low counts of GNR. Strong rancidity could be reproduced by adding the culture supernatant of a pre-grown lipolytic strain, but not the washed cells, to milk and pasteurizing it immediately before cheese-making, demonstrating the extracellular nature of the rancidity-inducing lipases.

Free fatty acids (FFA) and free δ-lactones in butter oil were quantitatively estimated by cold-finger molecular distillation at 35 °C followed by silicic-acid column chromatography of the cold-finger distillate and gas chromatography of the fatty acid and lactone fraction. Typical values for FFA and free δ-lactones in butter oil prepared from export standard sweet-cream salted Australian butter were in the range 0·07–0·24, 0·75–2·4, 4·5–8·6, 34–40 and 44–89 ppm for the C4,6,8,10,12 fatty acids and 0·09–0·18, 1·2–4·9 and 3·0–5·8 ppm for the C8,10,12 δ-lactones. These values are in the range of organoleptically desirable levels.

Phenol, o-methoxyphenol, m- and p-cresol, indole and skatole have been isolated from good quality butter oil by cold-finger molecular distillation, and separated into phenolic and indolic fractions by solvent extraction and silicic-acid column chromatography. The individual compounds were quantitatively estimated by gas chromatography. When phenol, o-methoxyphenol, m- and; p-cresol, indole and skatole were added to volatile-free butter oil, their recoveries were 94, 47, 90, 75, 71 and 61% respectively. The technique, when applied to fresh butter oil, gave the following ranges of values for the natural levels of these compounds: phenol 0·005–0·022, o-methoxyphenol 0·002–0·10, m-cresol 0·002–0·010, p-cresol 0·002–0·004, indole 0·07–0·13, skatole 0·16–0·22 ppm of butter oil. The results, when compared with flavour threshold studies, showed that indole and skatole are important contributors to the natural flavour of butter oil, but that phenolic compounds are of only borderline significance.