The proportions of milk stored in the gland cistern (cisternal) and within secretory alveolar tissue (alveolar) were determined for a group of ten cows in mid lactation, yielding ∼ 20 kg milk/d. As a proportion of total milk, cisternal milk, measured at 8 h after milking, ranged from 0·09 to 0·53. Milking frequency was then reduced from twice daily to once daily on two occasions, first for 1 week and then, 2 weeks later, for 2 weeks. Milk yield was reduced during once daily milking in all cows, by an average of 22·8% during the first week (P < 0·001). The amount of decrease varied among cows in a way that was unrelated to pretreatment yield, but the immediate decrease was significantly and inversely correlated with cisternal milk proportion (r = 0·81, P < 0·01). When milking reverted to twice daily, yield recovered to a value not significantly different from pretreatment, and the degree of recovery was inversely related to cisternal milk proportion (r = 0·74, P < 0·01). With the exception of one cow, yield decreases correlated well between the two periods of once daily milking. When treatment weeks were disregarded, lactation persistency (the rate of decline in yield with time) was unaffected by once daily milking. The results show that the ability of individual cows to tolerate once daily milking is related to their cisternal storage characteristics; yield is reduced less in cows that store a greater proportion of their total milk production within the cistern. Once daily milking for up to 2 weeks had no lasting effect on milk yield or lactation persistency, suggesting that there had been no detrimental effect on udder development.

Compensatory changes in daily milk production between quarters within an udder were investigated, together with related changes in cell count and electrical conductivity of fore milk samples. Two quarters per cow were left unmilked for 2 d (Group 1, four cows) or for 4 d (Group 2, four cows), and then all quarters of all cows were milked twice daily for a further period of 16 d. The mean daily yield per cow fell 50% during the period of milking two quarters. Daily yield for cows in Group 1 decreased significantly only during the treatment period and the first post-treatment period of 4 d, whereas the daily yield for Group 2 was significantly reduced during the whole post-treatment period. The cell count for all continuously milked (control) quarters was stable during the experiment, but increased in the treated quarters during the first 4 d of the post-treatment period. Non-milking was accompanied by a significant increase in fore milk conductivity which persisted for an additional 4 d after the resumption of normal milking. Throughout the last 12 d of the post-treatment period milk from both control and treated quarters had significantly decreased conductivity levels compared with the pretreatment values.

A technique for monitoring the movement of the teatcup liner during milking, using video recordings, has been investigated. The liner was videoed through a transparent shell and the recordings were subsequently processed frame by frame by an image analyser. The width of the liner and its variation with pressure difference across the liner walls were calculated at several locations along the liner length, and analysis of these results allowed the pressures at which the liner buckled and the walls touched to be determined and some estimate of the degree and duration of liner collapse to be made.

The proportions of milk stored within the gland cistern (cisternal milk) and within secretory alveolar tissue (alveolar milk) were determined at various intervals after milking in peak lactation cows and late lactation cows. The rate of milk secretion remained constant up to 12 h in both groups. Cisternal milk remained low (600 g or less) until after 4 h in both groups, then increased gradually to values at 12 h of 5·08 (peak lactation) and 2·60 kg (late lactation). Alveolar milk exceeded cisternal milk in both groups and at all time points up to 12 h, and was greatest in the peak lactation cows. A further measurement was made at 20 h in the late lactation group. Milk secretion rate remained constant between 12 and 20 h and most of this additional milk accumulated in the cistern. Alveolar fraction, defined as alveolar milk as a proportion of total milk, ranged from 0·43 to 0·94 at 8 h. There was a highly significant correlation between 8 h and 20 h alveolar milk fraction results, but not between 1 h and 8 h values, nor between alveolar milk at 0 h (effectively residual milk) and alveolar fraction at any time point. The milk volume results were used to construct a mathematical model describing milk flow between cisternal and alveolar compartments, and hence the pattern of milk accumulation. The results are discussed in relation to current hypotheses regarding local autocrine control of milk secretion by a feedback inhibitor in milk.

The effect of milking interval on secretion rates of milk and milk constituents was studied in four milking camels during late lactation. An interval of 4 h produced the highest milk secretion rate compared with 8, 12 and 16 h intervals, and rates tended to decline with increasing milking intervals. Secretion rates of organic (lactose, SNF, fat and protein) and inorganic milk constituents (sodium, potassium, calcium and magnesium) generally followed similar patterns of decreasing secretion with increasing milking intervals. Many of the values for different intervals were significantly different.

The transport of α-aminoisobutyric acid (AIB) by lactating rat mammary tissue has been examined. AIB uptake by mammary tissue was via both Na+-dependent and Na+-independent pathways. AIB uptake via the Na+-dependent pathway was inhibited by (methylamino)isobutyric acid (MeAIB) whereas AIB uptake via the Na+-independent pathway was blocked by 2-aminobicyclo[2, 2, 1]-heptane-2-carboxylic acid (BCH). A small fraction of AIB influx persisted in the presence of both MeAIB and BCH. The Na+-independent moiety of AIB uptake was strongly inhibited by phenylalanine, tryptophan, leucine, isoleucine and methionine. AIB efflux from mammary tissue slices was found to be both Na+-dependent and Na+-independent. Trans-stimulation of AIB efflux by other amino acids was not observed; in contrast, external phenylalanine, tryptophan and leucine inhibited AIB efflux. The results are largely consistent with the presence of systems A and L in lactating rat mammary tissue. However, the Na+-independent fraction of AIB transport may represent transport via a tissue specific form of system L.

The importance of elevated concentrations of oxytocin (OT) during the entire milking period was investigated in seven primiparous dairy cows with inherent disturbed milk ejection and in sixteen healthy cows with disturbed milk ejection induced by placing them in an operating theatre. Disturbance of milk removal in both groups has previously been demonstrated to be exclusively due to central blockage of the expected OT release in response to teat stimulation and milking. However, milk ejection can be induced by exogenous OT. OT (0·2 i.u.) was injected i.v. before milking and 49 ± 6% of the total milk was removed. When plasma OT decreased, milk flow stopped. In response to a second and third injection of 0·2 i.u. OT, 30 ± 4 and 7 ± 2% of the milk were removed respectively. The remaining milk was removed with 10 i.u. OT. The lag time from injection of OT to the start of milk flow was inversely correlated with the amount of milk actually removed in response to the OT injection. If 0·2 i.u. OT was injected during intramammary pressure (IMP) recording, IMP immediately increased to its maximum value. After 2·5 ± 0·3 min, IMP decreased to an intermediate IMP (between preinjection and maximum IMP). After two additional injections of 0·2 i.u. OT and after injection of 0·5 i.u. OT, IMP increased to a similar maximum. However, after injection of 0·5 i.u. OT, maximum IMP lasted longer (2·9 ± 0·3 min; P < 0·05) than after injections of 0·2 i.u. If OT was continuously infused (0·15 i.u./min) during milking, milk flow lasted until the udder was completely emptied. IMP increased during OT infusion to a maximum which remained stable until infusion was stopped after 10 min. The same IMP maximum was reached after the first individual OT injection (0·2 i.u.), but when plasma OT decreased towards basal concentrations, milk flow ceased and IMP decreased to an intermediate level. Thus continuously elevated OT concentrations such as those during infusion or during normal milking are necessary for complete milk removal.

In a 2 × 2 × 3 factorial design grazing experiment we investigated the effect of fertilizer (none or 240 kg N/ha), amounts of clover grass available (low or high) and type and level of daily supplementary feed for each cow (3·5 kg barley, 3·5 kg concentrate mixture rich in protein and fat, or both, 7 kg) on the protein composition and renneting properties of their milk. The experiment was carried out in two successive grazing seasons (years) and included a total of 79 Danish Holstein cows. The effect on milk protein composition was determined in both years whereas the effect on renneting properties was determined only in the second year. Fertilization of the clover grass significantly decreased total milk protein concentration (− 1·4 g/kg; P < 0·01) and tended also to decrease the relative proportion of whey protein N. Fertilization had no effect on renneting properties. Increased availability of clover grass significantly increased milk protein concentration (1 g/kg; P < 0·05) and resulted in significantly poorer renneting properties, that is increased clotting time (P < 0·01) and decreased coagulum development. These effects seemed to be mediated through an effect on the pH of the milk (+ 0·05; P < 0·05) as the effect was markedly reduced when statistical correction was made for the actual pH. Use of the protein- and fat-rich concentrate mix (3·5 kg) significantly reduced the total protein content of the milk (P < 0·05) and increased the proportion of non-protein N (NPN) in total N compared with use of the other supplementary feeds (P < 0·05). We found no effect on renneting properties of the different supplementary feeds. Throughout the grazing season and independent of the main treatments, the NPN proportion of milk N increased at the expense of casein N. At the same time, renneting properties became poorer, especially with high clover grass availability.

Changes in fat concentration and triacylglycerol fatty acid (TGFA) composition were studied in milk from six periparturient cows 1 d before and 20 d after an experimentally induced Escherichia coli mastitis in the fore and rear homolateral quarters. Opposite fore and rear heterolateral quarters remained uninfected and were used as controls. Milk was collected from all individual quarters during the experiment. The fat concentration in milk from infected quarters did not change, but total fat production decreased owing to reduced milk production after the Esch. coli challenge. In milk from the heterolateral uninfected quarters fat concentration rose significantly 48 and 72 h after induction of mastitis, the rise being concomitant with a decrease in milk production. Throughout the experiment similar changes in TGFA composition were observed for both infected and uninfected quarters. There was an increase in all the even, odd-numbered, iso and anteiso short-chain TGFA from day +6 on after induction of mastitis. There was little change in the composition of 16:0 and 18:0 fatty acids, while the long-chain unsaturated fatty acids decreased. Using multivariate analysis, the results are presented visually. The observed changes in the TGFA can be ascribed to changes normally observed in cows' milk soon after parturition.

Forty-four goats were milked and milk flow recorded without or with 1 min manual prestimulation in early, mid and late lactation. Ultrasound measurements of cross sections of the whole mammary gland were performed in a water bath. In additional experiments with 15 goats, milk flow was recorded and frequent blood samples were taken for the determination of oxytocin and prolactin concentrations. Milk yield increased from the first to the third lactation and decreased markedly during the course of lactation. Average and peak milk flow rates were closely related to the actual milk yield. The ultrasound cisternal area was 27·4±1·5% of the entire udder half cross section. Milking characteristics were scarcely different without or with prestimulation, although oxytocin was released within 30 s after the start of prestimulation, whereas oxytocin concentrations without prestimulation increased only after the start of milking. Concentrations of prolactin were higher during July and August than in April, and increased similarly with or without prestimulation during milking. In contrast to dairy cows, prestimulation and an opportune release of oxytocin during milking does not significantly influence the course of milk flow in goats, and this is probably because large amounts of cisternal milk allow milk ejection to be induced only after the start of milking without causing bimodal or otherwise reduced milk flow.

The whey proteins α-lactalbumin and βMactoglobulin have been investigated as potential markers of mammary development in sows by measuring their concentrations in plasma. The whey proteins were isolated from porcine milk by gel filtration, ion-exchange and hydrophobic interaction chromatography, characterized by several criteria and used to raise antibodies. Specific radioimmuno-assays were set up for porcine α-lactalbumin and β-lactoglobulin and validated for use in porcine blood and milk. Plasma levels of the whey proteins were measured in sows that were pregnant, suckling litters post partum, weaned abruptly at birth or were pregnant but mastectomized. Both whey proteins showed similar patterns in plasma post partum, falling from a maximum 1 d after parturition to values < 0·02% those in milk by day 4–5 post partum in suckling sows and showing a transient peak associated with early involution before declining to very low concentrations in non-suckling sows. α-Lactalbumin was first detected in the last week prepartum, rising markedly in the 3 d before parturition, correlated with rising prolactin (r = 0·986) and falling progesterone (r = –0·998). β-Lactoglobulin rose much earlier from 5 weeks prepartum, at the time when lobulo-alveolar mammary development is occurring, and correlated (r = 0·929) with oestradiol-17β. In mastectomized sows, concentrations of whey proteins in plasma were reduced by 90% or more when compared with intact animals, though following a similar pattern. This study shows that whey protein concentrations in plasma vary with physiological state and reflect aspects of the development of the mammary gland. The very different profiles for α-lactalbumin and β lactoglobulin prepartum indicate that they are differently controlled.

The transport of taurine by lactating rat mammary tissue has been examined. Taurine uptake was found to be dependent upon the presence of extracellular Na+ and Cl-, which is consistent with (Na+ + Cl- + taurine) cotransport. The Ka and Vmax of taurine influx were respectively 43 μM and 37·5 μmol/kg cell water per 15 min. It is apparent that the mechanism responsible for taurine uptake is highly selective for β-amino acids. Taurine efflux consisted of a fast extracellular component and a slow membrane-limited component. The slow component was relatively insensitive to temperature, suggesting that it may represent simple diffusion. Mammary tissue was found to contain a high level of intracellular taurine: 7·29–7·44 mmol/kg cell water. We suggest that taurine is taken up and concentrated by the mammary gland by a (Na+ + Cl- + taurine) cotransport mechanism situated in the blood-facing aspect of the secretory epithelium and that a low outward permeability to taurine allows a high intra-to-extracellular concentration gradient to be maintained. Milk taurine may be derived from taurine diffusing from the cell cytosol across the apical membrane.

The aim of this work was to investigate whether luteal oxytocin released after non-luteolytic prostaglandin F2α (PGF2α) challenge could explain the intramammary pressure (IMP) rises previously described in pregnant ewes after the normal life span of a corpus luteum. Blood oxytocin levels and IMP after challenge were measured in cycling and pregnant lactating ewes until the response ceased (˜ 55 d post oestrus). Oxytocin release was not significantly different for cycling and pregnant ewes, and fell to its lowest levels on days 16–18 post oestrus in both groups, i.e. with or without luteolysis. However, although IMP response fell with luteolysis in cycling ewes, the response persisted until day 55 in pregnant ewes. This lack of correlation between oxytocin release and IMP response raised interesting questions including: is there an unknown oxytocic compound that can be released after PGF2α stimulation of the corpora lutea?

An artificial teat containing a force transducer was constructed to study the forces exerted by the liner on the teat during milking. The objective was to develop a method of comparing the action of different liners under a variety of different milking conditions. The rigid structure of the artificial teat did not provide an accurate representation of a real teat but deformable models used in the past have not proved practical. Initial tests of the teat showed that it gave repeatable results during calibration. When placed in a liner, however, repeatable results could not be obtained if the teat–liner interface was disturbed. The forces that were measured under simulated milking conditions varied with liner wall position and with time. There was a peak and trough prior to a stable asymptotie value being attained and a noticeable time lag behind pulsation chamber pressure. This technique can be used to compare liners when a reliable method of locating the artificial teat within the liner has been developed.

The Malvern Particle Sizer, which applies Fraunhofer light diffraction theory, has recently become popular for measuring particle size distributions in dried milk and other powders. The size distribution is obtained by dispersing the powder in air or a suitable liquid in the laser light beam of the instrument. The suitability of the dry powder dispersion system (DPD system) of the Malvern Particle Sizer 2600c for characterizing agglomerated milk powders was examined. It was found that, although the instrument gave reproducible and reliable results for non-agglomerated powders (standard milk powders), milk powder agglomerates could be broken in the standard DPD system and erroneous size distributions might be obtained. However, the DPD system had potential for providing an objective test of the mechanical stability of milk powder agglomerates. In addition, the transient size distribution during dispersion of an instantized agglomerated milk powder in water was measured at intervals of about 20 s.

Mammary tissue from pigs on days 60, 80, 90, 100 and 100+ (days 106–111) of pregnancy has been cultured in vitro as explants. The total accumulation in tissue and culture medium of the whey proteins α-lactalbumin and β-lactoglobulin has been measured using specific radioimmunoassays. The control, uncultured tissue showed progressive morphological development from sparse, non-secretory epithelial tissue on day 60 to full lobulo-alveolar development with some accumulated secretion from day 100. In uncultured explants β-lactoglobulin could be detected consistently from day 90 (13 ± 12 ng/μg DNA, n = 4) and α-lactalbumin from day 100 (1·3 ± 0·5 ng/μg DNA, n = 11). At all stages of pregnancy, both whey proteins increased markedly during the period of culture (up to 7 d). Stimulation of α-lactalbumin appeared to be primarily under prolactin control. Prolactin increased α-lactalbumin accumulation to a similar extent alone, or in the presence of insulin and/or corticosterone. The response to prolactin was dose-dependent over the range 0·4–20 nM (10–500 ng/ml). Porcine prolactin was more potent than ovine prolactin. There was no effect of porcine growth hormone and no synergism detected between prolactin and tri-iodothyronine. By contrast, no specific hormonal requirements were established for accumulation of β-lactoglobulin, which appeared to increase in vitro if tissue remained viable in various combinations of insulin, corticosterone and prolactin. It was not stimulated by growth hormone. There was some indication of a prolactin-sensitive component in longer term cultures after day 4.

Radioreceptor binding studies were designed to localize and determine the number of α and β-adrenergic receptors in the mammary gland of lactating cows. 3H-prazosin, 3H-rauwolscine and 3H-dihydroalprenolol were used for the regional characterization of α1, α2- and β-adrenergic receptors by competitive inhibition of binding of 3H-ligands with unlabelled adrenergic agonists and antagonists. The α1-, α2- and β2-adrenergic receptor subtypes could thus be demonstrated in the regions of the teats, large mammary ducts and parenchyma. Tissues of the teat wall, of the large mammary ducts above the gland cistern and of the mammary parenchyma were prepared to determine the density of α1, α2- and β-receptors by saturation binding assays using 3H-prazosin, 3H-rauwolscine and 3H-dihydroalprenolol respectively. Binding to high affinity sites was reversible within minutes and saturable. Equilibrium was reached within minutes. The number of α1-and α2-adrenergic receptors decreased from the teat to the mammary ducts to the parenchyma. Most of the α1- and α2-adrenergic receptors were found in the teat wall, whereas in the parenchyma α-adrenergic receptors were absent or barely detectable. The density of β-adrenergic receptors was similar in the teat wall and the large mammary ducts, but much lower in the parenchyma. Thus, α1, α2- and β-adrenergic receptors were found mainly in the milk purging system and hardly at all in mammary parenchyma. Inhibition of milk removal by α-adrenergic stimulation is possibly due to constriction of teat wall and to constriction of the mammary ducts, whereas enhanced milk flow after β-adrenergic stimulation is possibly due to relaxation not only of the teat sphincter and teat wall, but probably also of the large mammary ducts.

The aim of this work was to test the diffusion-controlled hypothesis of milk coagulation kinetics by reducing the diffusion coefficient of casein micelles. This has been achieved by increasing the solvent viscosity of milk through sucrose addition. Milk was reconstituted from skim milk powder and sucrose added at 100–300 g kg–1. Hydrolysis and coagulation were followed by chromatographic determination of caseinomacropeptide content and optical, thermal and viscoelastic measurements. Soluble and ionic calcium were determined by atomic absorption spectrophotometry and ionometry and micelle size was measured by dynamic light scattering. Addition of sucrose resulted in a substantial retardation of both enzymic and aggregation steps, a re-equilibration of calcium because of water reduction, and a micelle size increase. The enzymic rate constant was inversely proportional to the viscosity, according to a diffusion-controlled model, and the lag or characteristic times for the aggregation reaction were inversely proportional to the viscosity. These results are consistent with the involvement of diffusion-controlled steps in the sequence of reactions.

Each of the secondary lipid oxidation products pentanal, hexanal and heptanal was found to react with β–lactoglobulin (β–lg) in a two-phase model System (aqueous phosphate buffer–1-octanol) yielding fluorescent condensation products (emission maximum, 410 nm; excitation maximum, 350 nm). Protein polymers were detected by size-exclusion HPLC, and the rate of reaction paralleled the formation of fluorescent products, with the reactivity being pentanal > hexanal > heptanal. Simultaneously, the reaction also changed the intrinsic fluorescence of β–lg, and in particular pentanal reduced the intensity of tryptophan fluorescence (emission maximum, 332 nm; excitation maximum, 288 nm) by 30%. These findings are discussed with reference to the effect of peroxidizing lipids on the physical properties of whey proteins and the use of protein fluorescence (induced by the reaction with aldehydes) as marker for the oxidative status of milk and whey protein products.

Whey clarification can be achieved by using a lipid aggregation step followed by microfiltration. The results from using an M5 Carbosep membrane to ultrafilter defatted sweet whey at pH values in the range 8·0–1·5 furnished understanding of the fouling process so that fouling may be minimized. The conventional method of aggregation, allowing the pH to decrease naturally, has been compared with a modified aggregation process in which the pH was maintained constant. These two methods differed significantly in their influence on the subsequent ultrafiltration (UF), with respect to the UF hydraulic characteristics, i.e. reversible, irreversible and overall fouling resistance. Optimal UF performance was obtained at a pH equal to or slightly higher than the aggregation pH (7·5) owing to the limited fouling contribution of proteins and calcium phosphates. The modified process permitted UF at fluxes in the range 50–115 1 h-1 m-2, with moderate transraembrane pressure, even with a protein content two to five times higher than that of regular whey.