Samples (120) of ewes' raw milk collected over a 12-month period on arrival at the dairy and freshly drawn ewes’ milk samples (30) collected at the farm were analysed for total viable counts and staphylococci. Total viable counts reached on the average 54 × 106/ml in dairy samples and 6·0 × 105/ml in farm samples, whereas mean levels of staphylococci were 5·9 × 104/ml and 3·3 × 103/ml, respectively. A high proportion (62%) of coagulase-positive isolates from Baird-Parker agar plates was detected in spring months. Staphylococcus aureus (42%), Staph. haemolyticus (23%) and Staph. epidermidis (10%) were the predominant species in 210 characterized isolates of staphylococci from ewes' raw milk. Seven coagulase-positive, maltose-negative staphylococci could not be identified with any of the recognized Staphylococcus species. Eighteen Micrococcus strains were identified as M. varians, whereas 111 Micrococcus strains of uncertain taxonomic status were separated into three groups, according to their physiological characters.

Lactoperoxidase (LP), thiocyanate (SCN-), pH and somatic cell counts (SCC) were measured in mammary secretions from 20 cows collected 14 d before drying-off, 7 and 21 d after drying-off, and 3–18 d postcalving. The inhibitory activity of the secretions on Streptococcus uberis was determined and the susceptibility of the udder to infection by this organism was tested by intramammary infusion of 250 colony forming units at the above stages. LP, SCN-, pH and SCO increased during involution and fell postcalving. The secretions collected before drying-off, 7 d after drying-off and postcalving inhibited growth of Str. uberis.; those collected 21 d after drying-off did not. Inhibitory activity in pre-drying-off secretions was destroyed by heating and restored by addition of LP, glucose and glucose oxidase, but addition of these substances to secretion 21 d after drying-off did not provide a full inhibitory system. The growth of Str. uberis in the secretions was correlated with intramammary susceptibility, since challenges with Str. uberis at 14 d before drying-off, at 7 and 21 d after drying-off and postcalving led to 43·8, 25·0, 81·3 and 37·5% of quarters becoming infected. It is suggested that the LP/SCN-/H2O2 system plays a role in protecting the lactating mammary gland from infection with Str. uberis but becomes ineffective as involution progresses.

Ewes' milk from two breeds of Greek sheep, Karagouniki and Serron, was analysed throughout mid and late lactation to investigate the salt distribution between colloidal and soluble phases. Separation of the phases was obtained by centrifugation at 48000 g. Effect of stage of lactation was significant only for total Mg in both breeds (P < 0·05) and for colloidal citrate in Serron ewe milk (P < 0·01). Breed differences were not significant (P < 0·05), except for colloidal casein.

In the colloidal phase there was a strong positive correlation of casein with Ca and P, but correlation with Mg was poor. Molar sums of [Ca + Mg] and [P + Cit] were also significantly correlated. The Ca to P molar ratio averaged 2·00 and was higher than that reported for cows' milk. The inclusion of Mg as well as citrate in the colloidal calcium phosphate is indicated. In the aqueous phase there was a strong negative correlation of Ca with pH and a positive one with citrate. Correlation with phosphate was less. It seems that citrate is the main factor influencing Ca concentration in the soluble phase.

The velocity of milk flow from the teat during the peak flow rate period of milking was estimated to be ∼ 8·5 m/s with a liner vacuum of 50 kPa and 7·5 m/s at 40 kPa. These results confirm the applicability of the Bernoulli theorem for estimating velocity of milk flow through the teat canal during milking. Concurrent measurements of mass flow rate and velocity indicate that the effective diameter of the open teat canal is ∼ 2 mm. Further calculations indicate that the shear force due to milk flow acting to debride the surface of the open teat canal is about 1·8 × 10-2 N. The opposing surfaces of the ‘closed’ teat canal may be ∼ 100 µm apart immediately after milking. If so, bacteria present in the milk residue within the teat canal would never be more than 50 µm from a surface.

The effect of subclinical mastitis on levels of plasminogen and plasmin in milk from cows in a high-yielding herd was investigated. Comparisons were made with levels of milk Na, antitrypsin and N-acetyl-β-D-glucosaminidase (NAGase). In samples from mastitic quarters plasminogen activity, as measured after activation to plasmin, increased by only 21% and plasmin by 82%, while NAGase increased by 307 %. Plasminogen was the only component that was normally distributed, all other components showed more or less skewed distributions. Plasmin and plasminogen were significantly related to the other components. However, plasminogen plateaued when the other components continued to increase. There was thus no further increase in plasminogen with the severity of inflammation as with the other components. Plasmin showed a similar although less pronounced tendency. Results of treatment of mastitic whey samples with acid suggested that the non-linear increase in plasmin activity was due to interaction with acid-labile proteinase inhibitors. Mastitis led to dissociation of plasminogen and plasmin from the casein micelles. The degree of activation of plasminogen was higher with casein-associated than with soluble plasminogen in both healthy and mastitic milks. Plasmin was very closely related to milk Na, which is a sensitive indicator of epithelial integrity. It is suggested that plasmin contributes to Na leakage into milk by degrading membrane proteins of the epithelial lining. Plasminogen and antitrypsin, which are both plasma proteins, were not identically affected by stage of lactation, indicating nonidentical modes of transport from plasma to milk.

The chymosin-initiated coagulation of casein micelles was followed by monitoring light transmission using a Platelet Aggregometer. The release of macropeptide by chymosin was monitored using fluorescamine. The lag period in the clotting reaction was proportional to clotting time and the reciprocal of enzyme concentration. The average rate of coagulation, which was approximately equal to the reciprocal of clotting time (Tc), increased in proportion to enzyme concentration at low enzyme concentrations and reached a limiting value at high enzyme concentrations. The percentage hydrolysis at the Tc was 47 ± 5% in the presence of 20 mM-CaCl2 and it was calculated that a 5-fold decrease in the speed of the enzyme-catalysed reaction would decrease this value at the Tc to 43 ± 5%. The possible uses and limitations of the Platelet Aggregometer for determining the influence of the chemical environment on the velocity of the chymosin-catalysed reaction and para-casein micelle aggregatability are discussed.

Mature lactating Friesland ewes had a mean lactation yield of 293 ± 26 kg during a lactation period of 35 ± 2 weeks giving an average daily milk yield of 1·2 kg/d. Ewes were injected intravenously after normal milking with either saline (sham) or oxytocin and then remilked to determine the volume of residual (alveolar) milk. After a long milking interval of 16 h oxytocin treatment gave a significantly greater total daily milk yield than the sham treatment (oxytocin minus saline, morning milking, 0·199±0·038 kg, mean ±s.e.m., P < 0·01). Oxytocin had a small significant reverse effect after a short milking interval of 8 h (afternoon milking, –0·065±0·022 kg, P <0·05). The average increase in total daily yield over four stages of lactation was 0·133±0·029 kg (P < 0·01), or 11% of the average daily milk yield. The galactopoietic effect of oxytocin was associated with the efficient removal of residual (alveolar) milk. Residual milk accounted for 7·4 and 27·2% of the total daily milk yield in ewes treated with saline or oxytocin respectively. Residual milk expressed as a proportion of daily total milk yield remained steady in ewes studied between April and July, but declined in September when yields were less than 1 kg/d.

Short-term associations between disease and milk yield were studied in 1594 lactations occurring between 1977 and 1982 in 732 Friesian, Ayrshire and Holstein crossbred cows. Statistically significant differences in milk yield between one week before and one week after clinical diagnosis were observed for ketosis (5·l kg/d), hypomagnesaemia (4·1 kg/d), mastitis occurring after peak yield (2·1 kg/d) and lameness (1·1 kg/d). Milk yield declined for as much as 2–4 weeks before diagnosis of disease, and total losses in milk yield associated with ketosis or hypomagnesaemia were estimated to be 60–70 kg. No significant differences were found for endometritis or for mastitis when it occurred before peak yield. Milk yield was suppressed for about 4 weeks after calvings with retained placenta, but there were no corresponding effects of dystocia or hypocalcaemia on milk yield. It is suggested that if automatic daily milk recording is available it may be possible to detect deviations from normal, and hence detect subclinical disease 2 or more weeks before its clinical appearance.

Coagulation of para-casein micelles was monitored using a Platelet Aggregometer at 37·8 °C and pH 6·7. The macropeptide released by chymosin was determined quantitatively with fluorescamine. The aggregatability of para-casein micelles integrated over the complete hydrolysis of κ-casein was calculated from the amount of κ-casein hydrolysed during the clotting time (Tc). Low concentrations of CaCl2 enhanced the rate of κ-casein hydrolysis but increases in CaCl2 concentration above 8 mM caused marked decreases in this rate. Calcium chloride enhanced the ability of para-casein micelles to aggregate. Little aggregation of para-casein micelles occurred at 0·6 mM-CaCl2 even after all of the κ-casein was hydrolysed. As the concentration of CaCl2 was increased from 3 to 60 mM, aggregation of para-casein micelles took place at progressively lower levels of κ-casein hydrolysis and the percentage hydrolysis at the Tc decreased markedly from 71±7% to 26±8%. The combined influence of CaCl2 on the velocity of the chymosin-catalysed reaction and on para-casein micelle aggregatability accounted for its effect on the average rate of coagulation (calculated by the reciprocal of the Tc). Results are consistent with the hypothesis that electrostatic repulsions and ionic bonding are involved in the interaction between chymosin and κ-casein.

The free fatty acid (FFA) levels after milking (initial FFA) and after 22 h storage at 4°C (FFA-22) were determined in milk from 57 cows every 2 weeks for one year. FFA levels which were higher in summer than in winter were explained by later stages of lactation occurring in summer. Initial FFA content increased regularly during lactation. FFA-22 content increased after 12 weeks, and further after 32 weeks of lactation. The first increase was due to stage of lactation. The second increase, which appeared in late lactation, was essentially due to the stage of pregnancy combined with the effect of milk yield. FFA-22 content increased in the second part of pregnancy, chiefly after 24 weeks. A low milk yield (≼ 5 kg) further increased lipolysis, but only after 16 weeks of pregnancy. The effect of stage of pregnancy may have been enhanced by management factors such as feeding conditions. In this investigation, the seasonal FFA variation in bulk tank milk could be explained entirely by physiological factors.

Bacterial growth was measured by a turbidimetric microtechnique in the whey of milk samples from quarters of cows with subclinical mastitis. Samples were grouped according to bacterial isolates recovered and the effects of bacterial species and whey on bacterial growth rates were analysed. Different strains of bacteria and different whey samples gave highly significant differences in bacterial replication rates. Except for penicillin-resistant Staphylococcus aureus, bacteria grew better in whey from mastitic milk where the inflammation was caused by the same bacterial species than in other mastitic milk samples. Inflammation caused by major pathogens generally enhanced the growth in whey of any type of major pathogen. Since mastitis pathogens showed enhanced growth in whey prepared from the same milk from which they were isolated, specific antibacterial factors in the whey did not appear to restrict bacterial growth in whey. The nutritional quality of the medium seems to be the important determinant of bacterial growth.

Half-udder comparisons were made using 56 cows for 2 months, in an experiment involving high bacterial challenge, to assess the combined effects of 5 min overmilking and pulsation failure (resulting from the use of shortened teatcup liners) on teat condition and mastitis. Only three new infections were confirmed in over 12500 quarter milkings in quarters milked with control liners (of 148 mm effective length) indicating little or no effect of prolonged overmilking in these quarters. A 3·5-fold increase in the new infection rate (NIR) based on bacteriological diagnosis alone (P < 0·01), or a 9-fold increase in NIR based on bacteriological diagnosis plus raised cell count and/or N-acetyl-β-D-glucosaminidase (P < 0·01), occurred in quarters milked with the same liners shortened to an effective length of 120–130 mm. The mean interval to infection or teat canal colonization (41 ν. 60 milkings), and for a quarter infection to be confirmed by other diagnostic tests (45 ν. 79 milkings) was significantly less in quarters overmilked with short liners. The results confirm that NIR increases whenever pulsation fails. Overmilking may increase NIR when it is associated with pulsation failure.

Several ionic components of ultrafiltrate were measured in bulk and individual cow milks and an assessment was made of their relationship with the parameters of the corresponding ethanol (EtOH) stability/pH profiles. From linear regression analysis the strongest relationships (P < 0·001) were between soluble salt balance [expressed as (Ca+Mg) minus (Pi+Cit) or as the ratio to (Pi+Cit)] and pK (correlation coefficient, γ ∼ 0·82) or Smax, the maximum stability at high pH (γ ∼–0·72), and between Pi and pK(γ = –0·84)or Smax (γ = –0·61). These relationships agree with the view that the parameters of the EtOH stability/pH profile are determined by pH-induced changes in concentration of divalent cations. Natural variations in these parameters may be attributed to variations in relative concentrations of divalent cations and their chelators. EtOH stabilities at the natural pH of bulk milks from winter/spring- and autumn-calving animals were lowest in early and late lactation. The most important contributory factors appeared to be a high salt balance ratio in late lactation and a low natural milk pH in early lactation. The main component responsible for variable salt balance ratio was usually soluble Pi. Decrease in EtOH stability at the natural pH of late lactation milks reflected a more general change in the characteristics of the EtOH stability/pH profile, seen as an increase in pK and, in extreme cases, a decrease in Smax and profile gradient.

The effect of aeration and agitation during milking on free fatty acid (FFA) levels in raw milk was investigated by comparing (a) a dual vacuum system with a ‘round the barn’ installation and (b) a claw piece requiring high air bleed with conventional claw pieces. Reduced milk transfer pressure resulted in a reduction in lipolysis of 54%, while high air bleed led to a 21% increase. FFA values were related to milk yield, stage of lactation and to a lesser extent the incidence of mastitis. These relationships were largely independent of the type of milking equipment. Fat content and somatic cell count did not affect the level of lipolysis. Increases in FFA due to activation by added blood serum were strongly related to the initial FFA concentrations.

Milk serum, whey and milk ultrafiltrate were examined by 31P nuclear magnetic resonance (31P-NMR). About 20 phosphorylated milk constituents gave rise to resonances in the spectra. Most of these have been assigned to such well-known milk constituents as inorganic phosphate, N-acetylglucosamine-1-phosphate and glycerophosphorylcholine. Resonances from previously unknown constituents such as phosphocreatine were also observed. When the pH-dependence of inorganic phosphate, N-acetylglucosamine-1 -phosphate, glycerophosphorylcholine and gly-cerophosphorylethanolamine was examined it was observed that the resonance of inorganic phosphate overlapped that of N-acetylglucosamine-1-phosphate around neutral pH. This is the most probable explanation as to why this constituent was not observed in earlier 31P-NMR studies on milk.

The effect of varying the casein composition of artificial micelle milk on rennet coagulation time and syneresis was examined in order to determine whether either of these processes is dependent on the concentration of particular casein components. It was found that the levels of κ-and β-caseins had a significant effect on coagulation, whereas syneresis was only affected by the level of β-casein. Partial dephosphorylation of preformed micelles or the incorporation of dephosphorylated or partly dephosphorylated β-casein into artificial micelle milk was found to have an adverse effect on both coagulation and syneresis. It was concluded that the phosphate groups of casein, particularly those of β-casein, are directly involved in the micelle-micelle interactions which occur during coagulation and syneresis.

Using either Sephacryl S-300 or TSK-G4000SW gel chromatography, the larger of the two major components of bovine casein micelles (F2) was eluted at the same retention time irrespective of sample concentration and column temperature. Its mol. wt was estimated to be 500K by examination of the eluate from a TSK-gel G4000SW column using a low-angle laser light-scattering photometer and a precision differential refractometer. With the second component (F3), retention time in Sephacryl gel chromatography was significantly affected by both sample concentra-tion and volume applied. Limiting retention times for F3 could be obtained by extrapolation of these two parameters to higher and lower values, and corresponded to those of water-soluble protein with mol. wt 200K and 100K respectively. Approach to sedimentation equilibrium technique at the above limiting sample concentrations gave values of F3 consistent with the above molecular weight. Sephacryl gel chromatography at either low sample concentration or low column temperature resulted in a new component being eluted at a position of mol. wt 25K

The chemical composition of Norwegian bulk collected goats' milk from the University herd was analysed during one lactation period (30 weeks, 20 samples during 1983). There was considerable variation in chemical composition during the year. Fat content decreased over the first 4 months of lactation and increased during the mountain pasture period. Protein concentration decreased during the first 4 months, and then increased until the end of lactation. Lactose concentration decreased throughout lactation. Casein nitrogen (casein N) was highest at mid lactation and lowest at the beginning and end of lactation. β-Lactoglobulin N showed the opposite trend. Citrate content showed a significantly quadratic decrease and total ash content an increase with advancing lactation. Mutual significant correlations between total P, K, Na, Ca and Mg were calculated, and all increased throughout lactation. There was significant positive correlation between concentrations of individual medium-chain fatty acids and stage of lactation. They remained more or less constant during the first part of the lactation, decreased to minima when the goats were on pasture, and increased during the last phase of lactation. Concentration of C16 fatty acid was negatively correlated with C18 and C18:1. Goat flavour intensity score and quality flavour score were highest at mid lactation, and positively correlated with the acid degree value.

Samples of reduced whole casein from genetically typed individual cows were quantitatively separated into their main components, αsl-, αs2-, β- and κ-caseins by reverse phase high performance liquid chromatography (RP-HPLC) using a mobile phase of phosphate-buffered aqueous propan-2-ol containing sodium dodecyl sulphate and an octadecylsilyl stationary phase. One casein sample was found to give two peaks of β-casein of approximately equal areas. RP-HPLC of tryptic digests of the two separated peak fractions gave identical patterns with the exception of one peptide peak with different retention times. Amino acid analyses performed on both fractions showed that they corresponded to peptide 114–169 of β-casein A1. The atypical β-casein differed from the typical one by a Pro→Leu substitution in region 114–169.

For milk from four herds of cows, maintained under different conditions of feeding and management, the natural Cu content and the stability of the ascorbate were highly correlated. Low Cu levels in milk from cows at pasture at farms A and B during the summer were associated with low storage losses of ascorbate. During this period, the milk of cows at farms C and D (on forage and Cu-supplemented concentrate) was richer in Cu, and losses of ascorbate were high. Heat treatment of the milk stabilized the ascorbate. Thus, in ‘high Cu’ milk (60 µg Cu/l), loss of ascorbate in the raw milk was 58% at 2 d, as against 17% after pasteurization at 72 °C and no loss after treatment at 82 °C. Storage of milk in light caused rapid destruction of ascorbate, equally with 72 and 82 °C heat treatments. The effects were examined of milk pasteurization temperature (72–82 °C) on flavour stability, bacteriological quality and vitamins of the B-complex. Heat treatment at 82 °C increased the susceptibility of vitamin B12 to destruction by light, but otherwise caused no greater losses of B-complex vitamins than did treatment at 72 °C. Taste panel ratings showed an initial preference for milk heated at 72 °C, but on storage of this milk in darkness the flavour score fell progressively and at 5 d it was judged ‘stale’. Treatment at 82 °C gave a faint ‘cooked’ flavour although, unlike that of the 72 °C-treated milk, the flavour remained stable throughout 14-d storage and after d 8 was increasingly preferred. On exposure to light after treatment at 72 °C milks rapidly acquired an unpleasant‘oxidized’ flavour, but after treatment at 82 °C, exposure to light had no such adverse effect on flavour during the early days of storage. Pasteurization at ∼ 80 °C offers a potential for improvement in the oxidative stability of milk and its contribution of vitamin C to the diet.