Three hundred and fifty-one lactating dairy cows, which were between 40 and 100 d post-calving, were selected from cows sampled during metabolic profile testing of 21 herds. Blood samples were analysed for packed cell volume (PCV), blood glucose and haemoglobin, and serum urea-N, albumin, total protein, inorganic phosphate, Ca, Mg, Na, K, Cu and Fe. After adjusting for herd and date of sampling and each cow's age, stage of lactation, milk yield and interval from calving to first service, concentrations of albumin and K (P < 0·01) and PCV (P < 0·05) were inversely related and concentrations of globulin (derived as the difference between total protein and albumin) directly related (P < 0·05) to the number of services required for conception. Conception rates to first service were inversely related to lactation number but not to milk yield on day of sampling.

In growing rats from 3 weeks to 3 months old, 0·1 % potassium thiocyanate (KSCN) decreased the plasma protein-bound iodine level by 47 % and the total DNA in the mammary gland by 11 %, but had no effect on the total KNA. Administration of thiocyanate in the form of 0·1, 0·3 and 0·5 % KSCN during pregnancy and 14 d of lactation lowered the protein-bound iodine level by 48–66 %, the total DNA by 11–47 % and the total RNA by 14–63 %, depending on the KSCN dosage. The 14-d-old litters of rats receiving thiocyanate weighed 4–41 % less than those in the control group.

Individual quarter milk samples were taken from 719 cows in 7 dairy herds and examined for mastitis-causing pathogens. Analyses were made to ascertain the prevalence of mastitis infection when the results from a single quarter were compared with those from all 4 quarters. There was a very close similarity between the 2 methods of assessment. When the number of samples is limited, sampling of a single quarter instead of all 4 quarters permits more cows to be examined.

The effect of intramammary infusion of Escherichia coli endotoxin upon mammary blood flow was studied in lactating goats and cows. Blood flow was recorded by means of an electromagnetic flow probe chronically implanted around one mammary artery.

Endotoxin mastitis was always accompanied by a significant increase in mammary blood flow, characterized by 2 conspicuous peaks. The flow returned to control values by the thirteenth hour after infusion. Other symptoms of acute mastitis were: fever, increased heart rate, swelling, heat and pain of the gland, increased chloride and total cell count in milk.

Using fistulated calves, the influence of the age, type of dietary protein and weaning on the secretion of chymosin and pepsin by the abomasum were studied. The abomasum secreted both chymosin and pepsin when the animals were fed milk. Chymosin secretion appeared to be independent of the age of the animals, whereas a slow increase in pepsin secretion was observed as the calves aged. Several preruminant animals were fed either a skim-milk diet or a milt substitute in which proteins were provided by fish, soya or whey concentrates. Each of these 3 milk substitutes led to a decrease in chymosin secretion without modification of pepsin secretion. Chymosin secretion was partly restored when the calves were again given a skim-milk diet. At weaning, chymosin secretion dropped abruptly, but the pepsin level was not affected. These results indicate that milk (most probably its casein fraction) is responsible for the activation of chymosin secretion.

Diagnosis of microbial infections in the udders of cows in commercial dairy farms for large experiments cannot be without error. Limitations of sampling method and routine prevent collection of the necessary information for sure diagnosis. However, with an organized method of repeated bacteriological examinations using consistent and proven methods of aseptic sampling the errors were shown to be very low. A method based on bacteriological tests on aseptic milk samples was used in 32 herds (approximately 2000 cows) for a 3-year period. This is described and examined in terms of other criteria to validate its use in experimental work. With this method it was not difficult to differentiate between those quarters which regularly shed pathogens and those which did not. Other evidence indicated that it was reasonable to assume that this classification accurately distinguished between infected and uninfected quarters. The errors using this method were quite small: when measuring the state of infection of all quarters in the herds the errors did not exceed 1 %. Some small modifications to the method described are suggested to improve further its diagnostic accuracy.

An experiment lasting 42 d was performed in 4 consecutive stages on 6 healthy Friesian cows during mid-lactation. Mean values for the different components of mammary secretion during normal lactation were established. Milking was then suspended on all quarters from d 1–14. The mean values for lactate increased 20- to 30-fold over the mean value for normal lactation. Over the same period the leucocyte count (polymorphonuclear leucocytes and lymphocytes) also increased, whereas the mean values for glucose and the estimated redox potential decreased. From d 15–28 milking was resumed on one half of the udder and from d 29–42 milking was also resumed on the other udder-half. When milking was resumed the above-mentioned changes were reversed, taking 5–7 d to reach values obtained during normal lactation in the udder-half which had not been milked for 14 d, but at least 14 d in the udderhalf which had not been milked for 28 d. The changes in the levels of lactate, glucose, leucocyte count and estimated redox potential are discussed in relation to changes in the secretory activity of the mammary gland (lactose, β-lactoglobulin and epithelial cell count), permeability changes of the mammary epithelium (Na+, K+ and serum albumin) and the immune defence mechanism in the udder (immunoglobulins). The results indicate that lactate is formed during anaerobic glycolysis by the leucocytes in the mammary secretion, most probably due to reduced blood flow to the udder and the accumulation of secretion in the gland and teat cisterns on cessation of milking.

A new procedure is described for the preparation of cytoplasmic droplets from goat's milk and the metabolic potential of these droplets has been surveyed. Acetate was incorporated into fatty acids of varying chain lengths and acetate and glucose were oxidized to CO2 by these cytoplasmic droplets in vitro. There was a low rate of protein synthesis but no detectable DNA or lactose synthesis. It is concluded that the cytoplasmic droplets can provide an alternative to biopsy material for studying certain aspects of goat mammary lipid metabolism in vitro.

A method for the quantitative analysis of casein mixtures is described. The method uses ion-exchange chromatography at 4 °C on DEAE cellulose, Whatman DE52 in the presence of tris-chloride-urea buffer (pH 8·6), and a NaCl gradient to fractionate the alkylated casein mixtures and a micro-biuret technique to determine protein concentrations. Values for fractions representative of (1) γ-caseins, (2) κ-casein plus some unknown proteins, (3) β-casein, (4) minor αs-caseins and (5) αs1- plus αs0-caseins were obtained, and whole casein from herd bulk milk was found to consist on average of about 2, 13, 36, 11 and 38% respectively of these fractions. Agreement between duplicate fractionations was satisfactory and the recovery of material varied between 94 and 98%.

The composition of whole casein of 5 herd-bulk and 42 individual-cow milks from a single herd of Ayrshire cows has been determined. The percentage composition of the herd-bulk samples varied relatively little over a period of more than a year with the whole casein consisting on average of 38·4 % αs1,0-casein, 36·5 % β-casein, 12·5 % κ-casein, 10·5 % minor αs-caseins and 2·1 % γ-casein. For the individual-cow samples from mid-lactation, casein composition also was relatively constant, but in early and late lactation composition varied more with the relative amount of β-casein often being lower and those of κ- and γ-caseins often higher than in mid-lactation, these differences tending to be more marked in samples from older cows. Stage of lactation appeared to have little effect on the relative amounts of αs1,0-and minor αs-caseins.

The concentrations of all casein fractions were significantly positively correlated (P < 0·001) with the concentration of whole casein in the milk. Also, there were significant negative correlations (P < 0·001) between the percentages of whole casein present as β- and κ-caseins, β- and γ-caseins and minor αs- and γ-caseins, and a significant positive correlation (P < 0·001) between the relative amounts of κ- and γ-caseins. The relative amount of the major αs1,0-casein fraction, however, was not closely related to those of any of the other fractions.

Generally, milks with high lactose and low Na values yielded casein which was comparatively rich in β-casein and comparatively poor in κ- and γ-caseins.

The efficacy of 18 disinfectant teat dips was tested on teats artificially contaminated with a milk suspension of Staphylococcus aureus. A solution of Na hypochlorite with 40 g/l available chlorine was significantly more bactericidal than one containing 1 g/l available chlorine and than most other disinfectants tested. The method was not able to distinguish differences in efficacy between solutions containing 40 g/l and 10 g/l available chlorine nor between these and some of the iodophors containing 5 g/l available iodine. The addition of 190–416 g/l (15–33 % v/v) glycerol significantly reduced the bactericidal properties of 3 iodophors (5 g/l available iodine), but soluble lanolin at approximately 20 g/l did not appear to lower the efficiency of NaOCl (45 g/l available chlorine) or of an iodophor (5 g/l available iodine).

Progressive changes in the concentrations of milk protein components were followed after infusions of Streptococcus agalactiae or bacterial endotoxin into different quarters of individual cows. Both types of infusion produced similar increases in somatic cell count and resulted in similar changes in milk proteins, although the effects of the endotoxin infusion lasted for a shorter length of time.

The treatments had little effect on α-lactalbumin and β-lactoglobin concentrations, but serum albumin and immunoglobulin (Ig) concentrations increased markedly. The greatest effect on serum albumin was after the endotoxin infusion and on Ig after the Str. agalactiae infusion. Changes in the individual globulins indicated that passive transfer of blood proteins to milk could not account for the observed increases in IgM and IgA.

αs1-Casein and β-casein concentrations were reduced and inversely related to somatic cell count during the immediate post-infusion period, and this was accompanied by an increase in para-κ-casein. Para-κ-casein was not detected in pre-infusion or post-recovery milk samples. The decrease in β-casein was greater than that of αs1-casein. Casein concentrations returned to pre-infusion levels 2 d and 5 d after the endotoxin and Str. agalactiae infusions respectively.

The possibility that proteolytic enzymes are partly responsible for the changes in casein concentration is discussed.

The components of whole casein from water buffalo (Bubalis L.) have been re-investigated in the light of recent studies elucidating the primary structure of all the bovine caseins (αs1, αs2, β & κ). Whole casein samples obtained from individual Italian buffaloes all contained 2 αs1- and 2αs2-casein fractions. These 4 components were isolated and identified by their amino acid compositions and terminal amino acid sequences. The 2 αs1-casein fractions seem to have identical peptide chains as do the 2 αs2 fractions, with the individual fractions within each group differing only in their phosphate content. Buffalo β-casein was also isolated and characterized by its amino acid composition and terminal sequences. The isolation of different buffalo κ-casein fractions was reported earlier (Addeo, Chobert & Ribadeau-Dumas, 1977). These fractions were purified and analysed. They were found to have the same amino acid compositions and C-terminal sequences, but to differ in their carbohydrate content. The prominent component, κ1, was treated with chymosin; the caseinomacropeptide and para κ-casein were separated from the digest and analysed. Starchgel electrophoresis at alkaline pH showed no polymorphism of αs1-, β- and κ-caseins in 170 individual buffalo whole casein samples. On 5 occasions the electrophoretic pattern of αs2-casein suggested genetic polymorphism. The behaviour of buffalo αs1−and αs2-caseins in the presence of Ca2+ was also studied.

Six adult West African dwarf does, about 2½ years old and weighing from 24 to 29 kg, were kept for 18 weeks/lactation, and for 2 lactation periods in all. During these periods the does were hand-milked twice daily and the daily samples were bulked for each animal for subsequent analysis. Milk yields for the period were low. Colostrum was characterized by a high content of total solids of 19·2%, fat 8·3%, total protein 5·1% and a gross energy of 517 kJ/100 g milk, but a low lactose content of 4·9%. The composition tended to approach that of mature goat milk between the second and third d after parturition. The mature goat's milk contained higher mean percentages of fat (6·9) and lactose (6·3) than any temperate breeds of goats and indigenous dairy cattle, but their protein (3·9%) and total ash (0·8%) contents were comparable with any temperate breeds of goats. The percentage protein, lactose, fat and total solids declined slightly with advance in lactation and there was a steady fall in milk yield which was statistically significant (P < 0·01).

This investigation was a first attempt to characterize the kinetics of the heat aggregation of the caseinate micelles in milk. The aggregation reaction subsequent to the beginning of visible aggregation in milk heated at temperatures in the range 110–140 °C appeared to be second order, with an activation energy of about 30 kcal/mol (124 kJ/mol). The kinetic parameters of the aggregation reaction were not significantly altered by the removal of serum proteins or of colloidal calcium phosphate, or by the inactivation of β-lactoglobulin.

Bovine milk-fat globule membrane (MFGM) has been fractionated in to 3 membrane components by isopycnic sucrose density gradient centrifugation. The 3 components, designated as heavy, medium and light material, were characterized by particular polypeptide, lipid, carbohydrate and enzyme compositions.

The heavy material (density > 1·145 g/ml) was red-brown in appearance and consisted of 63% protein, 32% lipid and 5% carbohydrate. The lipid moiety contained 40% triglyceride and 60% phospholipid, while the protein consisted of 3 major polypeptides (apparent mol. wt 135000, 70000 and 53000) in the ratio of 3:6:1. This membrane class was also characterized by the presence of both xanthine oxidase and acid phosphatase. The medium density material (density 1·055–1·145 g/ml) was light red to pink in colour and consisted of 35% protein, 58% lipid and 6% carbohydrate. The lipid contained 45% triglyceride and 55% phospholipid, with nearly 70% of the original MFGM phospholipid being found in this material. The ratio of the 3 major polypeptide bands was 2:4:4 and high levels of both alkaline phosphatase and 5'-nucleotidase were present. The light material (density < 1·055 g/ml) was white to pale pink in colour and was composed of 88% lipid of which 94% was triglyceride, together with 8% protein and 4% carbohydrate. There were virtually no enzymes present and 70% of the protein consisted of a polypeptide with apparent mol. wt of 53000. The 3 membrane classes also differed in carbohydrate content and in the fatty acid composition of their triglycerides.

When whole caseins from cow and Italian buffalo (Bubalus arnee) were fractionated by chromatography on a column of hydroxyapatite they behaved in a similar manner. κ-Casein was eluted with 5 mM phosphate buffer, pH 6·8, containing 0·2 M-KCI, 4·5 M-urea and 2 mM-2-mercaptoethanol, but β- and αs-caseins were retained and could be eluted successively by a linear gradient from 5 mM to 250 mMphosphate buffer. Buffalo κ-casein preparations, obtained from bulk milk or from milks of individual animals by chromatography on hydroxyapatite, produced identical electrophoretic patterns at pH 8·6. By further fractionation of these κ-caseins on DEAE-cellulose, in each case, at least 7 components were purified; they had different electrophoretic mobilities but were all sensitive towards chymosin. The major fraction migrated like component 1 of bovine κ-casein B.

Isoelectric focusing in thin-layer polyacrylamide gel has been applied successfully to the characterization and identification of calf rennet and its substitutes. The use of acidic pH gradients (pH range 2·5–6) allows the identification of calf and microbial rennets and bovine and pig pepsins.

A new, very rapid and sensitive zymogram technique has been developed by overlaying the gel with a highly porous, transparent cellulose acetate film impregnated with casein.

Additions of urea progressively increased the heat stability of milk outside of its coagulation time (CT)–pH minimum. In the region of the CT–pH minimum larger amounts of urea were required before an increase in heat stability occurred. The effect of urea was observed over the temperature range 125–140 °Cfornaturalmilk, milk which had been dialysed against synthetic sera, and milk to which a sulphydrylblocking agent had been added. Urea additions did not affect the activation energy of the heat coagulation reaction or the stability of milk to rennet or ethanol.