Pulsation rates of 40, 60 and 80 cycles/min were combined with pulsator ratios of 50, 60, 70 or 80% in two experiments with different liners. Machine-induced, short-term changes in teat thickness of 14 cows were compared with milk flow rate characteristics and machine strip yields to evaluate the effectiveness of pulsation in relation to liner type. Post-milking teat thickness increased progressively as the b phase of the pulsation cycle was lengthened, and as the d phase was shortened, at all pulsator rates and with either liner. Teat thickness values increased significantly (P < 0·05) when the d phase was <15%. For ratios of 50 and 60%, teat thickness decreased progressively as pulsation rate was increased. At the higher ratios, thickness values were lowest at 60 cycles/min. Pulsation settings that tended to increase teat thickness values also increased both peak milk flow rates and machine strip yields. The influence of liner type on teat thickness changes appeared to be at least as important as the influence of pulsator ratios and greater than the effect of pulsation rate. If so, then international standards for acceptable pulsation characteristics cannot be defined solely in terms of pulsator settings. A limit of ±5% for machine-induced changes in thickness of the teat apex would be an additional guideline for effectiveness of pulsation in relation to both liner type and vacuum level. This could provide a basis for a dynamic test applied to milking cows under field conditions.

For six successive milkings, six dairy cows were relocated immediately before milking to an unfamiliar operating theatre, a procedure previously shown to inhibit oxytocin release and milk ejection. Two control milkings were performed in familiar surroundings. After milk flow had ceased, two i.v. injections of 1 i.u. oxytocin were given to remove the remaining milk. Milk flow was recorded continuously and blood samples were taken every minute during milking and 10 min after milking. During the first milking in unfamiliar surroundings, no oxytocin was released. Thereby, only 13% of the total milk yield, the cisternal milk, was available and the alveolar milk fraction could only be removed after injection of oxytocin. During subsequent relocations oxytocin release steadily increased toward the control level, although the timing of oxytocin release remained delayed as compared with controls. However, the milk fraction available before oxytocin injection increased with increasing number of removals, following an asymptotic approach to control levels. The concentrations of β-endorphin, cortisol (and perhaps also of prolactin) gradually declined with the number of times the animal was moved to unfamiliar surroundings, i.e. hormone concentrations gradually adjusted to control level. During milking, concentrations of prolactin and cortisol increased, while β-endorphin concentrations decreased (except for the first relocation). We conclude that milking-related oxytocin release and therefore milk ejection adapted gradually to repeated relocations to unfamiliar surroundings. This adaptation was inversely related to β-endorphin concentrations, so it is possible that oxytocin release was suppressed by high circulating β-endorphin concentrations.

The effects of manual teat stimulation compared with stimulation by the liner of a conventional milking machine and the influence of milk ejection on the course of milk flow during the entire milking were investigated. Twelve cows were milked with or without prestimulation on three quarters and milk flow was recorded while intramammary pressure (IMP) was recorded in the left front quarter and blood samples for the determination of oxytocin (OT) were taken at 1 min intervals. OT concentrations increased very similarly in response to manual teat stimulation or liner stimulation and remained elevated during the entire milking procedure. Milk flow curves were generally bimodal without stimulation and not bimodal during milking after prestimulation. Milk yield was not significantly lower in milkings without than with stimulation, whereas machine-on time was prolonged and peak milk flow rate was reduced during milking without stimulation. Time to reach milk flow plateau, time to reach peak flow rate and IMP profiles were similar in both treatments if time was calculated from the start of stimulation. IMP increased similarly in response to manual or liner teat stimulation. In milkings with and without prestimulation IMP did not reach its maximum until the start of milking. In bimodal milk flow curves the second increase of milk flow occurred concomitantly with the start of IMP increase; ejection IMP generally occurred concomitantly with the milk flow plateau. In some cases during milking without prestimulation only small and transient OT release was observed. In response, IMP did not increase to its maximum and increased further in response to a stripping-related OT release. During the course of milking IMP decreased slightly because of the emptying of the three milked quarters. After the end of milking, and with OT concentrations decreasing to the premilking values, IMP again decreased slightly to another plateau, probably owing to myoepithelial relaxation. Thus the course of milk ejection, i.e. the IMP profile, influenced milk removal markedly during early milking.

Mammary gland protein metabolism, determined by an arteriovenous difference technique, was monitored in four Holstein-Friesian dairy cows in response to supplemental dietary protein (provided as rumen-protected soyabean meal) during late lactation (weeks 24–30). Each cow was offered two isoenergetic diets composed of grass silage (170 g crude protein/kg dry matter) plus either a low (108 g/kg) or medium (151 g/kg) crude protein concentrate in a single crossover design involving two 21 d periods. On day 21, arteriovenous measurements across the mammary gland were made during a 13 h continuous i.v. infusion of [1-13C]leucine and with frequent (2 hourly) milk sampling during the final 6 h. Although total milk yield was slightly increased (+1 kg/d) by protein supplementation, milk protein yield was not significantly affected. Whole body protein flux (protein synthesis plus oxidation) was not significantly affected by supplementation. Total mammary gland protein synthesis (milk plus non-milk protein) was also not affected by supplementation but on both diets gland synthesis was always greater (by 20–59%) than milk protein output. The fractional oxidation rate of leucine by the mammary gland was significantly increased by protein supplementation (0·047 v. 0·136). Although the enrichment of leucine in secreted milk protein continued to increase, the final value (at 13 h) was 0·94 of the arterial plasma free leucine plateau value (not significantly different), suggesting almost exclusive use of plasma free leucine for milk protein synthesis. Based on current feeding schemes for dairy cattle, a fixed proportion (0·65–0·75) of the additional protein intake (+490 g/d) should have been partitioned into milk protein. Instead, leucine oxidation by the mammary gland was increased. Whether oxidation of other amino acids was also enhanced is unknown but if amino acid oxidation and the ‘additional’ non-milk protein synthesis occurring in the gland are not crucial to milk synthesis, then by reducing such activities improvements in the efficiency of converting absorbed amino acid into milk protein can be achieved.

Four models describing renneting kinetics are evaluated for their ability to describe well documented attributes of the coagulation of casein micelles. The first model is based on a constant flocculation rate parameter. In the second the flocculation rate constant is proportional to the product of the sizes of the aggregating particles. Both models fail to predict proper dependence of rennet coagulation time on enzyme concentration. The third model is based on an energy barrier being reduced in linear proportion to the degree of proteolysis. The enzyme dependency of this model only works when the initial energy barrier is larger than ∼ 50 kBT (where kB is Boltzmann's constant and T the absolute temperature), which does not seem feasible. The fourth model, based on functionality theory, is able to predict proper dependence of rennet coagulation time on enzyme concentration when functionality is ∼ 2.

Preparation, physicochemical characterization and stability of iron-supplemented caseins were studied. When ferrous iron (final concentrations of 0·25, 0·5, 1·0 and 1·5 mM) was added to sodium caseinate (final casein concentration 25 g/l), iron binding to caseins was complete. In parallel, an increase in absorbance at 280 nm, a decrease in fluorescence intensity, modifications of reversed-phase HPLC profile, a decrease in pH and an increase in free sodium concentration were observed. Moreover, no release of iron from iron-casein complexes was found between pH 6·5 and 3·7 or after heat treatments or in the presence of inorganic phosphate. Only EDTA disodium salt and trisodium citrate had significant effects on the release of iron bound to caseins.

Monoclonal antibodies against peptidases of Lactococcus lactis were isolated and characterized: PEPN1–4 against a lysyl aminopeptidase PepN, PEPT1–5 against a tripeptidase PepT and PEPD1–3 against a dipeptidase PepD. These monoclonal antibodies reacted specifically with their respective antigens in crude cell extracts of Lc. lactis subspp. cremoris and lactis. A number of monoclonal antibodies cross reacted with proteins of other (lactic acid) bacteria. PEPT1, 2, 4 and 5 cross reacted weakly with a 35 kDa protein in Lactobacillus delbrueckii, while PEPT1 and PEPT2 reacted with proteins in the cell-free extract of Streptococcus thermophilus and Clostridium fervidus. Of the four isolated monoclonal antibodies against PepN, only PEPN3 cross reacted weakly with a 90 kDa protein in Escherichia coli cell-free extract, and the other three antibody species against PepN cross reacted with 80 kDa proteins of Lb. casei, Lb. delbrueckii, and Str. bovis, but not of Esch. coli. Of the three monoclonal antibodies against PepD, only PEPD1 and PEPD2 cross reacted with 40 kDa proteins of Lb. casei, Lb. delbrueckii and Str. bovis. All PEPN, PEPD and PEPT antibodies reacted with components in cell-free extracts of eleven different Lc. lactis strains, indicating that the peptidases of these strains were very similar to those of Lc. lactis subsp. cremoris WG2. However, Lc. lactis subsp. hordniae appeared to differ from the other Lc. lactis subspecies since only PEPT1, 2 and 5 reacted with a protein in the cell-free extract. Immunogold labelling of Lc. lactis WG2 with the isolated monoclonal antibodies revealed that PepN, PepD and PepT were located intracellularly. The intracellular location of these peptidases is discussed in relation to the supply of essential amino acids and peptides.

Studies on the immunomodulatory activities of ruminant milk and colostral whey fractions were undertaken. By comparing with boiled colostral whey in a preliminary experiment, a putative heat-labile immunostimulatory factor for antibody responses was found to be present in ovine colostral whey. Studies were then undertaken in sheep in which the efferent prefemoral lymphatic ducts were cannulated bilaterally, and immune responses in the node were measured following subcutaneous injection in the flank fold of whey protein preparations of various purities. A significant sustained decline of efferent lymphocyte output was observed following injection with autologous crude milk whey or colostral whey preparations, but no changes were observed in interferon-gamma levels in lymph plasma. Two bovine milk whey fractions (lactoperoxidase and lactoferrin) of high purity were compared in bilaterally cannulated sheep. A transient decline over the first 6 h was seen in the efferent lymphocyte output and lymph flow rate after injection of both fractions. A significant difference was seen between the two fractions in interferongamma levels in lymph at 6 h after injection. However, no significant changes in the proportion of the various efferent lymphocyte phenotypes were seen following either treatment. Whereas both fractions showed a significant inhibitory effect in a dose-dependent manner on the proliferative response of T lymphocytes, but not B lymphocytes, to mitogenic stimulation in vitro, no similar changes were seen following in vivo stimulation with these two fractions.

The effects of somatic cell count and stage of lactation on the yield and quality of Cheddar cheese were investigated. Cheese was manufactured in a pilotscale factory using milk of low bulk milk cell count (BMCC) from herds in early (LE) and late (LL) lactation, and milk of high BMCC from herds in early (HE) and late (HL) lactation. The deleterious effect of an elevated BMCC on product yield and quality in late lactation was clear. Cheese made from LL milk was significantly superior to that made from HL milk for most yield and quality characteristics measured. Stage of lactation also affected cheese yield and quality, as evidenced by the lower recovery of fat and poorer flavour score for cheese from LL milk compared with that manufactured from LE milk. The observed differences could be explained largely by differences in raw milk composition. We conclude that the effect of stage of lactation was magnified by an elevated BMCC, and that many of the problems encountered when processing late season milk could be overcome by containing mastitis at this time.

Skim milks were concentrated by ultrafiltration. Cream was added to the retentates to give cheesemilks standardized to a casein: fat ratio of ∼ 0·74 with protein levels ranging from 30 to 46 g/1. Pasteurized cheesemilks were renneted on a volume basis (22 ml single strength calf rennet/100 1) and converted to Cheddar cheese in 500 1 cheese vats. Set temperatures were reduced from 31 to 28 °C with increasing milk protein level to normalize curd firming rates and prevent curd shattering on cutting the curds. Cheesemaking was otherwise as normal for Cheddar cheese. The proportions of milk fat and protein lost in the cheese whey were not significantly influenced by milk protein level. Moisture-adjusted cheese yields increased with milk protein at a rate similar to that predicted by the Van Slyke cheese yield equation. However, owing to the negative correlation between cheese moisture and milk protein concentration, actual yields increased at a lower rate with respect to milk protein than moisture-adjusted yields. Increasing milk protein levels resulted in significant (P < 0·01–0·001) decreases in the concentrations of moisture, moisture-in-non-fat cheese solids and fat-in-dry matter in the cheese and increases (P < 0·05–0·001) in the levels of protein, salt-in-moisture, Ca and P. While increasing milk protein concentration resulted in significant (P < 0·05–0·01) reductions in the levels of water-soluble N at all stages of the 270d ripening period, it had little influence on the sensory scores awarded for aroma/flavour at 180 or 270d.

Eight strains of the Lactobacillus acidophilus complex were subjected, as concentrated cultures in skim milk, to different freezing treatments (rapidly in liquid nitrogen and more slowly in cold air at −30 °C), then stored at −80 and −30°C for 9 months. Survival was determined by plate counting with and without bile salts or sodium chloride at the highest tolerated concentration for each strain, to distinguish the undamaged population from the total population. Fermentative activity was measured as total lactic acid production by thawed cultures under standard conditions. Higher survival rates and greater activity were always obtained by storing cultures at −80 °C, but most strains stored at −30°C also survived well. Analysis of variance revealed that the viability of the frozen cultures was affected more by storage temperature than by cooling rate. Selective media were unable to distinguish the active population from the total surviving population. The correlation between values for activity and survival with selective media was poor.