The records of 453260 British Friesian cows, accumulated by the Milk Marketing Board of England and Wales in 1981/82, were used to estimate the effect of the calving interval on milk production in the subsequent lactation. There was a clear relationship between the length of the dry period and the calving interval in which it occurred, such that the dry period was extended by 2·5 d for every 10 d extension of the calving interval. The relationship between the interval and milk production in the following lactation appeared more likely to be due to variation in the dry period than to variation in the calving interval itself. A dry period between 50 and 60 d was associated with maximum production of milk, fat and protein. Among cows with dry periods shorter than 55 d, fat and protein production increased by 8 and 3·5 kg/d respectively. For cows with dry periods longer than 55 d, production of fat and protein was reduced by 0·2 and 0·12 kg/d respectively. The modal calving interval was 355 d including a dry period of 51 d. The interval tended to increase with the age of the cow, at the rate of 3 d/month of age within parity. The best estimate of future calving intervals was the mean of all previous intervals of the cow.

A New Zealand dairy herd of 73 cows was observed at pasture during the breeding season. Individual cow milk yields and composition analyses were obtained before, during and after oestrus. There was an overall reduction in yield at the first, and a rebound enhancement at the second milking after onset of oestrus. Changes in fat content followed the same time course and there was a slight increase in lactose content. Protein and somatic cell content were unaffected. There were considerable individual differences in both the duration and intensity of oestrous behaviour on the one hand, and the extent and timing of the effects on milk production on the other. However no relationship was found between the degree of behavioural excitement shown by a cow in oestrus and its change in milk production.

The yield and composition of milk from Friesian cows grazing either perennial ryegrass (Lolium perenne) (G, ten cows) or white clover (Trifolium repens) (C, nine cows) were evaluated between d 21 and 129 post partum. The two forages, of similar digestible energy content, were the sole source of nutrients and were offered ad lib. (exp. 1). Digestion and flow at the duodenum were measured on 13 occasions in early lactation with comparable cows fitted with rumen and duodenal cannulae and grazing similar forages (exp. 2). The gross milk yield (22·2, G; 25·0, C, kg/d P < 0·05) and the yield of protein (0·66, G; 0·77, C, kg/d, P < 0·01) were higher, but the protein content was similar and the fat content lower for cows fed C compared with G. Cows fed G were heavier at the beginning of the experiment and lost weight more rapidly than cows fed C. Milk energy output, adjusted for tissue energy gain or loss, was 83·9 for cows fed C compared with 71·8 MJ/d for cows fed G (P < 0·001), during the period of tissue energy repletion (weeks 11–18). From week 18 to the end of lactation all cows from exp. 1 were fed silage ad lib. and 502 kg dry matter of concentrates. The total (305 d) difference in lactation response to grazing C compared with G was 931 1 (5657 1, C; 4726 1, G); this was a direct response during the experiment of 301 1 and a residual response of 630 1. In exp. 2, more organic matter (6·47, G; 8·01, C, kg/ d, P < 0·001), and more non-ammonia N (433, G; 575, C, g/d, P < 0·001) entered the duodenum of cows grazing C compared with G.

Milks from two groups of dairy cows grazing either white clover or ryegrass swards were compared to determine whether their acceptability in terms of flavour, composition and rennet coagulation properties was affected by these differences in the cows' diet.

The concentrations of Ca and Mg were slightly higher and that of inorganic phosphorus was slightly lower in milks from the animals on the ryegrass pasture compared with those grazing clover, but the levels of Na, K and citrate were the same. The clover milks contained more αs- and β-casein and a greater proportion of small micelles but had similar contents of κ-casein to the ryegrass milks. Following addition of rennet at pH 6·4, the clover milks gave rise to firmer curds but the clotting times and rates of syneresis were not consistently different from the ryegrass milks. The clover milks contained a greater proportion of short-chain (C4–C10) fatty acids, but lower proportions of long-chain (C18+) and unsaturated acids than the ryegrass milk.

The different pastures did not give rise to any differences in the flavour of the milk.

The chemical composition and rennet coagulation properties at pH 6·4 of bulked milks from two groups of dairy cows grazing ryegrass or white clover swards were monitored over a period of 14 weeks commencing at the fifth week of lactation. There was considerable variation in the concentrations of many components and large variations in the rennet clotting time (RCT), coagulum strength and rate of syneresis. The relative proportions of αs-, β- and κ-casein were also subject to considerable variation. Many statistically significant correlations were found between concentrations of protein fractions, minerals and citrate, some of which may be of physiological significance whereas others may be incidental. The coagulum strength displayed significant correlations with concentrations of total casein, αs-casein, inorganic phosphate (Pi), whey protein and RCT, but not with β-casein or η-casein. The syneresis time was significantly related to the concentrations of fat, citrate, η-casein and the original milk pH. RCT displayed a significant correlation with the concentration of Pi.

The 31P nuclear magnetic resonance spectrum of liquid milk was examined. Of the three peaks observed, the two larger were assigned to inorganic phosphate (Pi) and the seryl phosphate (SerP) residues of casein; the third peak was assigned to a phosphodiester, which is probably glycerophosphoryl choline. The pH-dependences of the chemical shifts of the Pi and SerP were measured with and without added EDTA, and the results confirm the assignments. The width of the Pi peak in milk is significantly greater than in similar solutions lacking casein, probably because of binding to, or chemical exchange with, the casein micelle. Most of the SerP residues in milk are not sufficiently mobile to have been detected in these experiments but a significant fraction of SerP residues is able to move freely and can be titrated.

The increase in levels of free fatty acids (FFA) in morning and afternoon milk after 48 h cold storage at 5 °C was determined at two stages of lactation in milk from 96 cows. Lipolysis was positively correlated to day of lactation (r = 0·6, P < 0·001), while the correlations between lipolysis and milk yield and fat content respectively were due to these factors being related to day of lactation. In afternoon milk the level of FFA increased almost three times above that in morning milk. Lipolysis was closely related to the lipoprotein lipase (LPL) activity associated with the cream fraction (r = 0·8, P < 0·001). Afternoon milk had higher LPL activity in the cream fraction than morning milk. Lipase activity in the cream increased with increasing stage of lactation. Cooling of milk increased LPL activity associated with the fat phase. This effect was greater in afternoon milk from late lactation than in morning milk from early lactation. When heparin was added to milk the LPL activity associated with the fat increased substantially; again, this effect was greater in afternoon milk from late lactation than in morning milk from early lactation. Lipolysis was higher in heparinized milk than in normal milk, and there was a close correlation between lipolysis and LPL activity associated with the fat (r = 0·82, P < 0·001). Characteristics of the milk fat globule influencing the attraction of LPL were found to be fundamentally important to lipolysis.

The increase produced by formaldehyde (HCHO) in the heat stability of milk did not occur when milk was treated with HCHO at temperatures up to 60°C followed by dialysis at 5°C. However, the minimum in the heat coagulation time (HCT)–pH curve was irreversibly removed if the milk was preheated at 80–C for 10 min in the presence of 5 mM-HCHO. Although this treatment blocked the ε-amino groups of lysyl residues, the stabilizing mechanism is considered to be due to the cross linking action of HCHO which reduced the level of non-sedimentable, κ-casein-rich protein dissociated from the micelles on heating. The specific crosslinking agent, dimethyl suberimidate, modified the HCT-pH profile of milk in a manner similar to preheating at 80°C for 10 min with 5 mM-HCHO, supporting the crosslinking hypothesis. The results of this study appear to lend some support to the proposal of Kudo (1980) that the minimum in the HCT-pH curve of milk is due to the dissociation of κ-casein from the micelles on heating at high temperatures at pH values greater than 6η7.

Pseudomonas strain AFT 21 produced three heat stable extracellular proteinases in milk and nutrient broth at 7 or 21 °C, but the proportions depended on medium and cultivation temperature. The three proteinases were EDTA- and o-phenanthroline-sensitive metalloenzymes and were not inhibited by N-ethylmaleimide or phosphoramidon. Proteinases I and II showed maximum activity at pH 7–7·5 and proteinase III at pH 8·5. All three enzymes showed maximum activity at 45–47·5 °C, but had relatively high (19–27% of maximum) activity at 4 °C. They were unstable at 55 °C in phosphate buffer, pH 6·6, or synthetic milk ultrafiltrate (SMUF) containing 12 mmol Ca2+, but were stabilized by short preheating at 100 °C. They were extremely heat stable in both phosphate buffer and SMUF, pH 6·6, at 70–150 °C. Their D-values at 140 °C were 69, 54 and 80 s respectively. The Z-values for Pseudomonas AFT 21 proteinase III in phosphate buffer and SMUF were 29·7 and 30·3 °C respectively; the corresponding activation energies for inactivation were 8·7 × 104 J mol-1 and 9·2 x 104 J mol-1.

The effect of chelating agents on extracellular proteinase production by Pseudomonas fluorescens 32A was examined. Increasing concentrations of orthophosphate slightly stimulated growth while inhibiting proteinase synthesis. Fifty percent inhibition was found at 35 and 28 mM-orthophosphate at 5 and 20 °C respectively. Extracellular protein concentration was reduced by 30% when cells were grown with 100 mM-orthophosphate. Polyacrylamide gel electrophoresis of the cell-free supernatants suggested that reduced enzyme synthesis had taken place as evidenced by the decrease in staining intensity of the protein band corresponding to the proteinase. Other phosphate compounds could replace orthophosphate as an inhibitor. Extent of inhibition was related to chain length; polyphosphates with 4–6 or 13–18 phosphorus atoms were the most effective inhibitors. EDTA (0·5 mM) completely inhibited proteinase synthesis. This inhibition could be partly reversed by Ca2+ and, to a lesser extent, Mn2+. Proteinase production at 5 °C in skim milk was completely inhibited by phosphate glass (P13–P18). Control experiments showed that loss of activity with chelators was not due to inhibition of preformed enzyme. The results suggest a possible role for polyphosphates in controlling proteinase production in stored milk.

The caseinolytic activities at pH 6·8 of polymorphonuclear (PMN) and mononuclear leucocyte homogenates (equivalent to a level of 106 cells/ml milk) were less than the levels of natural milk proteinase activity found in milk from healthy cows. Bulk milks contained ∼ 4 times more milk proteinase activity than the composite milks from individual healthy cows. Isolated blood leucocytes, when added to raw milk of good bacteriological quality and stored at 5 °C, did not readily degenerate and had no detectable effect on the milk proteins even when these cells were completely disrupted by homogenization of the milk. Pasteurization of milk which contained leucocytes caused loss of cell vitality. Extracellular proteinases of psychrotrophic bacteria growing in milk were not detected until the early stationary phase of growth. The total viable count at which this occurred varied greatly. Proteinase production by a pure culture of Pseudomonas fluorescens was not detected in milk stored at 5 °C until a viable count of ∼ 109 colony forming units (c.f.u.)/ml was obtained, whilst normal bulk milks stored at 5 °C produced detectable levels of extracellular proteinase(s) when the psychrotrophic flora reached 107–108 c.f.u./ml. Casein proteolysis by PMN and mononuclear leucocyte homogenates resulted in similar polypeptide maps, but plasmin and bacterial proteinase isolated from a strain of Serratia marcescens resulted in polypeptide maps different from each other and from that produced by the leucocyte proteinase(s). The rate of proteolysis of caseins by the different proteinase sources appeared to be in the order αsl- > β- > > κ-casein for the leucocyte extracts, β- > αsl- > > > κ-casein for bovine plasmin and β- ≈ κ- > αsl-casein for S. marcescens proteinase.

A bacteriophage, øC2(W), which attacked Streptococcus lactis strains C2, ML3 and 712, is described. It had a prolate head and a non-contractile tail and produced large haloes around plaques. Infection of eight paired-strain cultures, each containing Str. lactis C2, with øC2(W) resulted in marked inhibition of acid production for six cultures. Direct phage–nonhomologous ‘host’ contact was not required for inhibition. Lysates of øC2(W) contained a phage-induced lysin. Evidence is presented that the inhibitory effect of øC2(W) against paired and multi-strain cultures is due to the lysis of phage-insensitive strains by phage lysin. An isometric phage, ø712, was shown not to produce such a lysin.

The lytic enzyme present in øC2(W) lysates was isolated by means of ion-exchange chromatography and further purified by gel filtration and ultrafiltration. The phage enzyme had an apparent pH optimum of 6·5–6·9 and was rapidly inactivated at temperatures > 47°C. The apparent temperature optimum was 37°C and Q10 and Ea values over the range 22–32°C were 2·5 and 69·2 kJ/mol respectively. Monovalent and divalent cations activated the enzyme. Reduced -SH groups on the enzyme were required for lytic activity. Gel filtration revealed a mol. wt of ˜ 46000. Strain-dependent differences in sensitivity of group N lactic streptococci to lysin were found. Group D streptococci were also lysed. Strains of three species of Leuconostoc, two species of Lactobacillus, one strain of Escherichia coli and of Micrococcus lysodeikticus were apparently resistant. Analysis of cell wall degradation products gave results which were consistent with the lysin having the specificity of an N-acetylmuramidase.

To obtain information about the possible release of β-casomorphins from β-casein under in vitro conditions, cows' milk was incubated with 13 strains of Gram-negative or Gram-positive bacterial species isolated from bovine milk. After incubation periods of 1–24 d, milk samples were assayed for β-casomorphin-4, -5, -6 or -7 immunoreactive materials. In general, no β-casomorphin immunoreactive material was found in samples incubated with non-caseolytic strains, e.g. Pseudomonas putida or Streptococcus faecalis subsp. faecalis; however, high levels were found in several samples incubated with caseolytic strains, e.g. Ps. aeruginosa or Bacillus cereus. When such materials were present, the amounts of β-casomorphin-4, -6 or -7 greatly exceeded β-casomorphin-5 immunoreactivity. It was concluded that while some of these materials probably represented authentic β-casomorphins, others were more likely to be β-casomorphin precursors.

The involvement of toxic oxygen intermediates in the bacteriostatic effect of milk was determined by producing bacterial growth curves using turbidimetry in the presence and absence of oxygen radical-scavenging substances. Using whey as substrate, catalase, haemoglobin combined with ascorbic acid and xanthine oxidase inhibitors all provided protection against oxygen toxicity for a strain of Staphylococcus aureus and of Streptococcus agalactiae. Superoxide dismutase and mannitol were less effective. This was evident in whey alone and in the presence of oxygen radicals produced exogenously by the t-butylhydroperoxide, H2O2 and xanthine/xanthine oxidase systems.

The effect of preheating skim milk and artificial micelle milk on curd syneresis was studied. The inhibition of syneresis caused by heat was dependent on the presence of β-lactoglobulin (β-lg) and to a lesser extent α-lactalbumin. The degree of inhibition increased with increasing amounts of added β-lg and preheating temperature. This agrees with the hypothesis that the detrimental effect of preheating on syneresis is due to complex formation between β-lg and κ-casein. This complex appeared to be mediated via thiol–disulphide exchange and its formation appeared to interfere with the micelle–micelle interactions responsible for syneresis.

The use of a silica-based ion-exchange medium for the recovery of protein from wheys prepared by treatment with rennet or with acid has been investigated. The protein capacity of the medium whilst maintaining an adsorption greater than 75% during the passage of rennet-and acid-whey was 0·079 and 0·053 g/g respectively. Bovine serum albumin (BSA), α-lactalbumin (α-la) and β-lactoglobulin (β-lg) adsorbed from acid-whey were mostly recovered in yields greater than 93% in an undenatured form essentially free of lactose and milk salts by elution using 0·1 M-HC1. With rennet-whey, BSA and part of the α-la were not recovered in the native form, probably because of proteolysis during elution by enzymes originating in the rennet used in the manufacture of the whey. There was, however, no effect on the recovery of native β-lg adsorbed by the medium. An unidentified proteinaceous fraction, thought to be of casein origin, was also adsorbed and recovered from rennet-whey. Fractions of the adsorbed proteins could not be obtained by eluting with a gradient increase in NaCl concentration. Nevertheless, purified β-lg was obtained by passing a large quantity of whey through the medium. This protein had a high affinity for the functional groups of the medium, to the extent that other proteins initially adsorbed were subsequently displaced. Unidentified proteinaceous material in rennet-whey also had a high affinity for the functional groups of the ion exchanger but most of this fraction could be removed by a pretreatment procedure which involved passing the whey through the medium at pH 5·0; unidentified material was selectively adsorbed at this pH. The medium could also be used to produce a fraction containing a mixture of BSA and α-la but it was not possible to separate these proteins.

A commercial neutral proteinase was encapsulated in liposomes (multilamellar) and incorporated into cheese curd via the cheese milk. The milk proteins were protected from proteinase attack during curd manufacture. A significant proportion of the liposome-entrapped enzyme was retained in the curd and its subsequent release into the cheese matrix was inferred from an increased rate of β-casein breakdown.

The effects of sorbic and propionic acids on shelf life and organoleptic characteristics of West African soft cheese stored at 7–9 °C were investigated. In cheeses treated with 0·1% sorbic acid spoilage was delayed and microbial growth, especially mesophilic bacteria, yeasts and moulds, was suppressed compared with untreated controls. Treatment with 0·05% sorbic acid or up to 10% propionic acid was less effective. An objectionable bitter flavour was reported for sorbate-treated cheese but other qualities were not adversely affected.

Yogurt, made from fortified skim milk by conventional methods using Streptococcus thermophilus and Lactobacillus bulgaricus, was used in studies of the effect of fermentation on nutritional value of milk. In all experiments, the product was compared with the uninoculated base milk. The concentration of most vitamins was less in yogurt than in milk and was most noticeably so for biotin which was 60% less. The effect on folic acid content was inconsistent. In nutritional experiments with rats, high values for true digestibility, biological value and net protein utilization were obtained for both yogurt and its base milk, only minor differences being apparent between the two materials. In growth tests with rats, yogurt was not found to be consistently superior to the base milk when the milk was subjected to a double heat treatment to reduce bacterial contamination. Yogurt did not confer a nutritional advantage on fresh milk in this respect.