The effects of the isocaloric replacement of starch in a low-fat concentrate mixture by either 5 or 10% ‘stearic acid’ (85% pure) or by 10% ‘palmitic acid’ (85% pure) on the composition of the plasma lipids were investigated in a feeding experiment with 12 cows in mid-lactation. The concentrate mixtures were given with a high roughage diet that supplied daily 4·4 kg of hay and 2·7 kg of sugar-beet pulp. A study was made of the relationships between the compositions of the plasma and milk lipids.

The inclusion of 10% ‘stearic acid’ or 10% ‘palmitic acid’ in the concentrate mixture increased the concentration of total plasma fatty acids. Irrespective of dietary treatment, about 40% of the total plasma fatty acids occurred in the cholesteryl ester fraction, 54% in the phospholipid fraction, 3% in the triglyceride fraction and 3% in the unesterified fatty acid fraction. There was a positive curvilinear relationship between the concentration of unesterified fatty acids in the plasma and the yield of total milk fatty acids.

In the plasma triglycerides, the concentrations of 16:0 and 16:1 were decreased and the concentration of 18:0 was increased when the concentrate mixture contained ‘stearic acid’; the concentration of 16:0 was increased and the concentrations of 18:0, 18:1 and 18:2 were decreased when the concentrate mixture contained ‘palmitic acid’. Similar changes were observed in the compositions of the plasma unesterified fatty acids when the cows were given the different diets.

In the plasma cholesteryl esters, the concentration of 16:0 was decreased and the concentrations of 18:3 and 20:4 were increased when the concentrate mixture contained stearic acid; the concentrations of 16:1, 18:3 and 20:4 were increased and the concentration of 18:2 was decreased when the diet was supplemented with palmitic acid. The addition of stearic acid to the diet increased the concentration of 18:0, 18:1 and 18:3 in the plasma phospholipids but decreased the concentrations of 16:0, 18:2, 20:3 and 20:4. When the diet contained palmitic acid the concentrations of 16:0, 16:1, 18:1 and 18:3 in the plasma phospholipids were increased but the concentrations of 18:0, 18:2 and 20:3 were decreased.

The major fatty acid circulating in the plasma of the cows was 18:2, which accounted for about 45% of the total plasma fatty acids. Only about 0·7% of the total plasma 18:2 occurred in the plasma triglycerides.

The results are discussed in relation to the changes in the composition of the milk fatty acids produced by the cows when they were given the experimental diets.

A lactating mammary gland of a sheep and a goat were perfused for several hours in the presence of [U-14C]D-fructose and received adequate quantities of acetate, glucose and amino acids.

In both experiments, there was a small incorporation of 14C in the expired CO2. Smaller radioactivities were measured in milk citric acid, lactose, casein and fat, the activities decreasing in that order. The specific activities of the amino acids from one casein hydrolysate were determined. The highest radioactivities were found in alanine and serine; methionine, glutamic acid and aspartic acid showed a smaller incorporation of 14C.

These results indicate that fructose is metabolized only to a very limited extent by the mammary gland. The relative distribution of 14C observed among the different substances isolated may be explained by a direct splitting of fructose into two C3-fragments, glycolysis and metabolism via the Krebs cycle.

Cows were given either a low-fat concentrate mixture or a concentrate mixture that contained 5 or 10% ‘stearic acid’ (85% pure) or 10% ‘palmitic acid’ (85% pure). The concentrate mixtures were given with a high roughage diet that supplied 4·4 kg of hay and 2·7 kg of sugar-beet pulp/day. Blood samples were taken from the cows on each dietary treatment and the plasma phospholipids were analysed.

Phosphatidyl choline accounted for 70% of the plasma phospholipids when the cows were given the low-fat diet and about 86% of the plasma phospholipids when the cows were given the diets supplemented with the fatty acids. The inclusion of the fatty acids in the concentrate mixtures decreased the relative proportions of the plasma phosphatidyl ethanolamine, sphingomyelin and lysophosphatidyl choline but the relative proportion of the plasma phosphatidyl serine remained unchanged.

When the diet was supplemented with stearic acid the concentrations of 18:0 and 16:0 in the phosphatidyl choline were unaltered but the concentration of 18:1 was increased and the concentration of 18:2 was decreased. When the diet was supplemented with palmitic acid the concentrations of 16:0 and 18:1 in the phosphatidyl choline were increased and the concentrations of 18:0 and 18:2 were decreased. In contrast, the inclusion of stearic acid in the diet increased the concentration of 18:0 in the phosphatidyl serine and decreased the concentration of 16:0; the concentrations of 18:1 and 18:2 were unchanged. The fatty acid composition of the plasma phosphatidyl ethanolamine was unaffected by dietary treatment. The effects of diet on the fatty acid compositions of the phosphatidyl choline and lysophosphatidyl choline were similar. Supplementation of the diet with stearic acid increased the concentrations of 18:0 and 18:1 in the plasma sphingomyelin and decreased the concentrations of 16:0 and 18:2. The addition of palmitic acid to the diet increased the concentration of 16:0 in the sphingomyelin but it decreased the concentrations of the other constituent fatty acids.

Aseptically drawn milk was made into small cheeses (100 g) under aseptic conditions. The influence of the cow's feeding régime on the composition of the cheese fat, and the hydrolysis of the cheese fat and protein by bacterial and native milk enzymes, was studied.

In the absence of a strongly lipolytic bacterial cheese flora, the free fatty acid (FFA) content of cheese appeared to depend on several variables:

1. The initial FFA content of the milk varied according to the mode of milking (the lowest was obtained by cannulation of the udder) and the feeding régime of the cow. When one of a pair of monozygotic twin cows was maintained on pasture and the other on winter feed (hay and concentrates), the cheese made from the milk of the latter contained less FFA than did that from the former.

2. The major changes in the composition of the milk fat of the cows on winter regime were an increase of myristic and palmitic acids from 8 to 11% and 21 to 28%, respectively, and a decrease in stearic and oleic acid from 11 to 7·5% and 38 to 28%; minor changes in the other fatty acids were also observed.

3. The milk lipase in raw milk cheese liberated substantially more FFA than did the weakly lipolytic lactic acid streptococci. Milk lipase was active at the low pH value of cheese, but was inactivated when the milk was pasteurized at 63°C for 30 min.

4. Although rennet did not produce any amino acids in the mature cheeses it hydrolysed some of the cheese protein to nitrogen soluble in water (WS), trichloroacetic acid (TCA-S) or phosphotungstic acid (PTA-S). Some of the WS-N appeared to be utilized by the lactic acid streptococci. The native milk protease resisted the heat treatment used and liberated amino acids at the low pH value of cheese, but its contribution to the proteolysis was relatively less important than the contribution of the milk lipase to the fat breakdown in cheese.

5. The free amino acids of the milk, which appeared in the whey, were not recovered in the cheese. The cheese made from unheated milk with δ-gluconic acid lactone contained low amounts of free amino acids apparently due to the action of milk protease. The proteolytic activity of the lactic acid bacteria increased the amino acid content appreciably.

The isoprenoid fatty acid 4,8,12-trimethyltridecanoic acid has been isolated from butterfat and identified. This acid was found to be a DD diastereoisomer, and was thought to have been derived from the phytol moiety of chlorophyll. It was estimated that in the sample of butterfat investigated, 4,8,12-trimethyltridecanoic acid constituted about 0·005% of the total weight of fatty acids.

Curd was made in a continuous curd-making apparatus (Berridge & Scurlock, 1969) and immediately separated from its whey. Its temperature was maintained at a high level (41–48 °C) as it was moved continuously in the form of a thin layer through a set of rollers. The curd then had a moisture content similar to that of normal curd at the end of scalding in the traditional process. After a suitable period of cheddaring the pH value and the moisture content of the continuously made curd were the same as those normally reached at the same stage in the traditional process.

(1) A survey of the composition of the milk of individual cows in 4 commercial herds in the West Riding of Yorkshire was undertaken from October 1967 to May 1968.

(2) In 3 of the 4 herds there was a decline in solids-not-fat (SNF) content throughout the winter feeding period. Milk protein content was at a minimum in January–February but in all 3 herds lactose content also declined and in 2 of the herds this was the major cause of the fall in SNF content.

(3) The decline in lactose content was the result partly of the characteristic effect of an advance in lactation, partly of an increase in the incidence of mastitis throughout the winter period. In all herds there was a significant inverse relationship between lactose content and cell count in the bulked milk of individual cows.

(4) In 3 of the herds autumn-calving (October–December) animals showed a pronounced fall in milk protein content in the early months of lactation which was not observed in summer-calving (July–September) animals. There was no response in SNF or protein content to supplementary concentrates offered towards the end of the winter-feeding period, whereas with the commencement of grazing in the spring there was an immediate increase in both protein and SNF contents.

Rennin hydrolysed the phe-met bond in the peptide H-ser-leu-phe-met-ala-OMe (i.e. methyl ester), the amino acid sequence of which is similar to that around the phe-met bond attacked by rennin in κ-casein. Rennin did not attack other peptides from this sequence not containing serine, and it is suggested that, in both κ-casein and the pentapeptide, the enzymic attack is accelerated by the nearby serine side chain. Rennin also hydrolysed sulphite esters such as phenyl sulphite ester and some N-substituted imidazole compounds such as benzoyl imidazole. Phenyl sulphite esters may be suitable substrate for assaying the activity of preparations of rennin.

An antibacterial agent is produced by ultraviolet (UV) irradiation of Streptococcus cremoris strain C11–56. Its production is accompanied by cell lysis and it attacks the strain C11–56 which produces it as well as a number of other Str. cremoris strains. A correlation between sensitivity to the antibacterial agent and sensitivity to trypsin has been demonstrated. Electron micrographs have shown the presence of particles resembling phage heads, most of which are empty, and of only very few intact phage-like particles in the lysate. The active agent in the lysate is associated with a particle of M > 100000. The evidence suggests that the activity of the lysate may be due to the presence of a defective phage or of a lethal component of it.

In correctly operated continuous heat-treatment plant the processing temperature is maintained at a sufficiently elevated level to give the desired thermal death rate for any micro-organisms present before treatment. This paper shows that if through imperfect operation of the plant, the processing temperature fluctuates about the required level, the chances of survival for bacteria and spores will theoretically be increased. Tabulated results show the magnitude of the effect under varying conditions and the compensating increase in processing temperature to counteract it.

The dynamic viscosity coefficients η of creams containing up to 50% fat have been determined at temperatures of approximately 40, 50, 60, 70 and 80°C. All the creams behaved as Newtonian liquids, the shear stress being proportional to rate of shear up to the maximum rate used of 100 sec−1. At a given temperature a linear dependence of log η on (φ+φ5/3) was obtained for φ < 0·4, where φ is the concentration (w/w) of fat. Interpolation formulae have been derived to enable η to be calculated at any temperature between 40 and 80°C and for any fat content up to 40%. Formulae for the density ρ of cream have also been deduced to permit kinematic viscosities η/ρ to be computed. Nomograms have been constructed to enable η and ρ to be readily determined without the use of the interpolation formulae and when slight loss in accuracy is unimportant.

The nature of the αs-casein complex of bovine milk has been investigated by cation-exchange chromatography on columns of sulphoethyl Sephadex C-50 of purified preparations of αs-casein and of whole acid-precipitated casein. Three main fractions were separated from each. Two behaved as single homogeneous proteins as judged by starch-gel electrophoresis; they were the main constituent of the complex, αs1-casein, and a closely related phosphoprotein, designated αs0-casein, which was present in small amounts only. The third fraction also constituted only a small part of the total complex. It was heterogeneous on starch gel electrophoresis and contained 2 major and 2 minor components. This fraction, while similar to other members of the αs-casein complex and to β-caseins hitherto described in that it contained phosphorus, nevertheless differed significantly from these since 3 at least of its components contained sulphur either in the form of disulphide bonds or of sulphydryl groupings.

Information is presented on the composition of the 3 fractions including C-terminal end group analysis.

In experimental small-scale manufacture of Cheddar cheese a study was made of the movement of lactose, lactic acid, calcium and phosphorus between curd and whey, and of the effect of high acidity developed in the curd before whey separation, on the pH and physical properties of the cheese. It was found that while the curd particles remain in contact with the whey, the lactose fermented in the curd is replaced by lactose diffusing from the whey. Lactic acid produced in the curd diffuses into the whey rather slowly.

When a high lactic acid level is attained rapidly the curd does not cheddar well, the pH of the cheese is low, the flavour is sour and bitter, the body is crumbly and the cheese is bleached. When a high lactic acid level is reached slowly, the curd also does not cheddar properly but the pH of the cheese may be within normal limits up to 3 weeks, at which age its body is coherent and its colour is normal. However, on maturing, the cheese becomes acid, the body crumbly and liquid separation occurs even at 2 months of age. This points to a risk in grading cheese at an early age.

It is suggested that the abnormal behaviour in cheddaring of ‘high-acid’ curds is connected with an excessive loss of calcium. When high acidity is developed over an extended time in the whey, the curd retains more lactose but loses more phosphorus so that its buffering capacity is reduced. Cheese made from such curd becomes highly acid at maturity and this leads to the development of a bitter flavour.

It is, therefore, concluded that both the rate of development of acidity, and the time the curd is in the whey, are critical factors in controlling the quality of Cheddar cheese.

The milk-clotting and proteolytic activities of rennet, bovine pepsin and porcine pepsin were compared. The milk-clotting activity of porcine pepsin was extremely pH-dependent around pH 6·6 and coagulation did not occur above pH 6·68. The clotting activity of bovine pepsin was slightly more dependent on pH than that of rennet but no rapid drop-off in activity occurred as with porcine pepsin. Temperature influenced the clotting activity of rennet and bovine pepsin similarly but the behaviour of porcine pepsin was markedly different.

For equal milk-clotting activity, the proteolytic activities of rennet and bovine pepsin were approximately equal and substantially lower than that of porcine pepsin. Electrophoretic examination showed that the proteolysis products of rennet and bovine pepsin were similar and quite different from those produced by porcine pepsin.

The suitability of bovine pepsin as a rennet substitute is discussed.

Differences in milk-fat composition in summer and winter have been demonstrated and related to feeding for an autumn calving Friesian herd and an Ayrshire herd in which calving was equally divided between spring and autumn. The observations are in agreement with previous results obtained for milk fat in the Northern Hemisphere.

Summer milk fat contained more C18 acids and less C4–C16 acids than winter milk fat, the effect being particularly marked for the autumn calving herd.

Changes in the pattern of feeding altered the fatty acid composition of milk fat from both herds, and abrupt changes in feeding were clearly reflected even at times when stage of lactation effects were greatest.

Effects of age and agitation of milk, temperature of separation, heat treatment and cooling, homogenization and ageing on the body of double cream were studied. Homogenization and temperature of heat treatment gave rise to the greatest effect. The effects of most treatments were inter-related to some extent, but this was most pronounced with cooling and ageing.

Using bacterial endotoxin as a tracer material, it has been shown that a jet of liquid impinging on the end of the teat for 1 min after milk flow had ceased during machine milking could force material past the barrier of the streak canal. Positive results were obtained when the maximum pressure at the centre of the impact area was about 0·25 bar, with a considerably higher frequency of positives at 0·3–0·5 bar. It seems possible that such impact pressures might occur during milking and cause transfer of infective material to the teat sinus. Impact force could arise when milk returned to the teatcup liner as the result of local or general conditions of instability in the milking machine vacuum.

The method of Gaitonde (1967) was adapted for the measurement of cysteine and cystine in acid hydrolysates. Analysis of 3 different casein preparations indicated the presence of both cysteine and cystine in whole casein in the proportions of 1:1 by weight and in the molecular ratio of 2:1. If all the cysteine and cystine is associated with κ-casein then it appears that one mole of the latter contains 2 moles of cysteine and 1 mole of cystine. The sulphydryl group in whole casein appears not to be highly labile.

The folate in cow's milk was strongly and specifically bound to a minor whey protein (FP), forming a complex of primary M of about 38000, but exhibiting concentration-dependent reversible aggregation. The binding protein was present in excess, and the milk had the capacity to bind about 50 μg added folic acid/l. An enriched concentrate of FP was prepared by ammonium sulphate fractionation—FP was precipitated at between 50 and 60% saturation—and further purified by chromatography in DEAE-cellulose and filtration in Sephadex gel G150. Its identity as a distinct minor whey protein was confirmed by comparative starch gel electrophoresis at various pH values.

Some properties of the protein are described, and its physiological significance discussed.

Loss of milk clotting activity when rennin was treated with urea at several concentrations and at different pH values was compared with the change in UV absorption of the protein. The change in absorption at 287 and 293 nm did not correlate with loss in activity, indicating that the loss of enzymic properties was not related to the extent of unfolding of the molecule but was related to the reaction of susceptible groups to the environmental conditions.

Reactions of modifying agents, for example N-bromoacetamide, 2-hydroxy-5-nitrobenzyl bromide and mercuric ions with tryptophan groups and the relationship of these changes to enzyme activity suggested that tryptophan was not specifically associated with enzyme action but was important in maintaining tertiary structure.

Rennin was not inhibited by the pepsin inhibitors p-bromophenacyl bromide and α-diazo-p-bromoacetophenone which suggested that the active site did not possess a reactive carboxyl group as in pepsin. However, the reversible inhibition with 1-cyclohexyl-3(2morpholinoethyl)-carbodiimide metho p-toluenesulphonate together with the results of oxidation by N-bromoacetamide indicated that a tyrosine group or groups was important in enzyme activity.