The effects of milking cows with two different liners were measured for a period of 8 months with 115 Danish Holstein cows divided into two groups. Group H and L animals were milked with liners with mouthpiece cavity heights of 30 and 18 mm respectively (other dimensions also differed between the two liners). Average teat lengths of first lactation cows were 45 and 40 mm for front and rear teats. Older cows had teats ∼10 mm longer. There was no difference in milk yield or milk flow rates between the two groups. Average machine-on time was shorter for group L, and first lactation cows of group L were less restive. The frequency of red and blue discoloured teats immediately after milking was higher for group H, and teat length increased on average 5 mm during lactation with no increase for group L. The small overall differences in udder health between the two groups were not significant. Udder health was better for first lactation cows of group L, even though liners of group L slipped more often and cows with recorded liner slip had poorer udder health. We conclude that special attention should be given to first lactation cows when liner type is selected for a herd. We propose that breeding programmes should ensure that teat length is kept above 50 mm.

The effects of reducing the frequency of milking of cows in late lactation on milk somatic cell count (SCC), polymorphonuclear leucocyte (PMN) content, chemical composition and proteolytic activity were investigated. Intermittent milking is frequently practised by Irish farmers in late lactation, and the objective of this study was to determine whether this procedure could be linked to altered quality of milk. Seventeen Holstein Friesian cows in late lactation (>215 d in milk) were assigned to two treatment groups, and were either milked twice a day until drying-off (control group) or milked intermittently as the yield fell (test group). Milk composition and enzymic characteristics were measured on two occasions. At the first sampling, day 7, test cows were on once daily milking and at the second, day 15, the test cows were being milked every second day. Milk yields were significantly lower in test than control animals and decreased between days 7 and 15 in both groups. Milk SCC and PMN levels were increased on reducing milking frequency and, at day 15, the increase was not linked to decreased milk yield. Milk lactose levels were significantly decreased and pH, α-lactalbumin levels, plasmin activity and plasminogen activity significantly increased by reducing milking frequency. In conclusion, reduced frequency of milking in late lactation leads to the production of milk that is abnormal in character and this may be linked to reduced quality of dairy products manufactured from such milk.

The aim of this study was to identify and rank the various factors, in particular those involving feeding, that affect the proportion of caseins in milk true protein. Twenty-nine feeding trials involving 821 lactations were assessed, and lactoprotein genetic variants were known for 551 of these. The main factor affecting the casein: protein ratio was the genetic variant of β-lactoglobulin: once corrected for other factors, the milk of BB type animals had a ratio nearly 30 g/kg total protein higher than AA animals. κ-Casein variant B also had a positive effect (+12 g/kg in favour of BB relative to AA animals). Except in the last weeks of pregnancy and the first weeks of lactation, the casein[ratio ]protein ratio varied little during lactation. It was significantly reduced when milk cell count exceeded 200000 cells/ml, even in the absence of clinical mastitis. It also decreased slightly with parity. Among the various dietary factors studied (level and type of nitrogen and energy supplies, forage type and preservation method), none had any significant effect on the milk casein[ratio ]protein ratio, except in drastic dietary situations. That ratio increased very slightly in parallel with the animals' milk yield and milk protein content. In practice, measuring the milk protein content in animals free from clinical mastitis remains a very precise predictor of casein content, accounting for 93% of its variation.

The effect of the proportion of clover in the diet (200, 500 or 800 g/kg total dry matter (DM)) on milk production of cows housed indoors and fed on a mixture of perennial rye-grass and white clover was measured in mid (Expt I) and late (Expt II) lactation. Higher clover contents increased the nutritive value of the diets, resulting in increased energy and protein intakes. DM intakes of cows offered 500 or 800 g clover/kg DM diets ad lib. (Expt I and Expt II, Period 1) were not significantly different but were 11–17% greater (P<0·05) than intakes of cows fed on 200 g clover/kg total DM diets. Cows offered restricted allowances (Expt II, Period 2) had similar intakes irrespective of diet. In Expt I cows fed on 500 or 800 g clover/kg DM diets ad lib. produced 30 or 33% respectively more milk (P<0·05) than cows fed on 200 g clover/kg total DM diets. During Expt II, Period 1, cows fed on 500 or 800 g clover/kg DM diets ad lib. produced 18 or 16% more milk (P<0·05) respectively than cows given 200 g clover/kg total DM diets. In both these experiments the increased milk yields were due to increased intake and the higher nutritive value of the high clover diets. There was no difference in the feed conversion efficiencies of cows if maintenance energy requirements were taken into account. However, cows on restricted allowances (Expt II, Period 2) showed no significant difference in milk yield, indicating that the effect of increased nutritive value was very slight. There were no consistent effects on milk fat, protein or lactose concentrations. Concentrations of blood and milk urea increased as the clover content of the diet increased (Expt 1 only), and this was associated with increased milk non-protein N and a decreased ratio of casein N[ratio ]total N. Both trials indicated an optimum clover content in the diet for milk production of 600–700 g/kg total DM.

This study was designed to evaluate the respective influences of stage of lactation (SOL) and time of year on the seasonal variation in milk composition for pasture-fed dairy cows in New Zealand. Four herds of ∼20 Friesian cows were used, one herd calving in a 6 week period beginning in each of January, April, July and October. Cows grazed rye-grass–white clover pasture only, except during June when all cows received supplementary pasture silage. Milk samples were collected from each cow in milk on four occasions during the year (September, December, March and June), to give a total of three samples per cow (early, mid and late lactation; about 30, 120 and 210 d respectively after calving). Samples were analysed for a detailed range of components. Concentrations of many milk components (e.g. total protein, fat, casein and whey protein) increased as lactation progressed; the extent of these increases depended on the time of year. These results indicated that spreading calving throughout the year would lessen seasonal variations in the gross composition of milk supplied to factories, leading to a more even distribution of product yield across the year. Despite this, variations in some important manufacturing properties were affected by time of year but not by SOL. Ratios of protein[ratio ]fat and casein: whey protein were not significantly affected by SOL, but were affected by time of year. The solid fat content of milk was also affected by time of year. Seasonal variations in the manufacturing properties of milk may be reduced but not eliminated by changing the time of calving.

The effect of graded levels of dietary rumen-protected methionine (rp-Met) or lysine (rp-Lys) on the fatty acid composition of Comisana ewes' milk was investigated. The animals lambed in autumn and were separated from their lambs 7 weeks after parturition; they were divided into five groups of nine which were given different rations. The control group was fed on a mixture of vetch and oat hay with a pelleted concentrate; the other groups were given the control ration supplemented with rp-Met (3·5 or 7·0 g/kg) or rp-Lys (10·5 or 21·0 g/kg). Milk fatty acid composition was determined by gas chromatography, and these values were subjected to principal components analysis. A correlation was found between fatty acid composition and diet. We demonstrated that giving the experimental diets significantly affected milk fat, to an extent that was independent of the levels of the supplementary amino acids. The milk fat of the control group contained largely caprylic, capric and lauric acids, whereas that of the groups given the supplementary amino acids contained largely palmitic, palmitoleic, stearic and oleic acids. Thus supplementation resulted in an increase in the proportion of 16[ratio ]0–18[ratio ]3 (primarily palmitic and stearic) and a reduction in the proportion of 4[ratio ]0–12[ratio ]0 fatty acids. There was also a modest increase in the unsaturated: saturated fatty acid ratio.

Our aim was to determine the effect of growth hormone on non-insulin-mediated glucose disposal in lactating dairy cows. Following 5 d of subcutaneous injections of either saline or growth hormone, insulin, somatostatin or insulin plus somatostatin were infused for 2 h each, in a series of experiments. Coincident with this, unlabelled glucose was infused at a variable rate to maintain a constant plasma glucose concentration. Glucose, doubly labelled with deuterium, was also infused for the calculations of glucose turnover. Plasma insulin levels were reduced to nearly zero by the infusion of somatostatin; under such conditions whole body glucose disposal should be non-insulin-mediated. Dairy cows treated with growth hormone, which had significantly increased milk yields on the day before the experimental infusions, did not have different levels of whole body non-insulin-mediated glucose disposal when expressed in absolute terms. Growth hormone did not affect non-mammary non-insulin-mediated glucose uptake estimated by calculation. Growth hormone significantly inhibited insulin-mediated glucose uptake when plasma insulin levels were elevated. Glucose uptake during insulin plus somatostatin infusion was not significantly different from that of the insulin only infusion.

Plasma concentrations of glucagon-like peptide-1(7–36)amide (GLP) and gastric inhibitory polypeptide (GIP) were determined at fortnightly intervals for over a year throughout the pregnancy–lactation cycle of goats. Both GIP and GLP concentrations were elevated during lactation and fell rapidly when milk secretion was terminated. At the onset of lactation GLP concentrations rose rapidly whereas GIP concentrations showed a delayed response. GLP concentrations remained high throughout lactation but those of GIP declined linearly as milk yields fell. Serum insulin concentrations correlated positively with plasma glucose concentrations but not with either GIP or GLP concentrations. Negative correlations were found between serum insulin concentrations and milk yield and plasma non-esterified fatty acid concentrations. The results are consistent with plasma GIP and GLP concentrations being determined by other factors in addition to nutrient intake and absorption. Changes in GIP concentrations mirrored reported changes in the hypertrophy and atrophy of the intestine in ruminants while GLP concentrations may be more dependent on the neural and endocrine factors associated with lactation. The elevated concentrations of both peptides indicated a specific role in lactation independent of their normal anabolic and insulinotropic effects.

Microfiltration (MF) of milk with a ceramic membrane (0·1 μm mean pore diam.) for the separation of casein micelles from soluble proteins was studied. Experiments were performed at constant flux density (J=65 or 75 l h−1 m−2) and wall shear stress (τw=100 or 110 Pa) with milks containing particles and solutes with different sizes and charges, produced by physicochemical change (heat and mechanical treatment, pH, ionic strength, addition of ions). Membrane separation performance was limited by the build up of a cake with characteristics that depended on the size of particles and soluble proteins. Best performance (higher permeability and whey protein transmission) was obtained with milk containing fat on the one hand and calcium phosphate on the other. An optimal value of the particle size was found, close to 0·5–3·0 μm; above this separation performance decreased. In addition, the present study confirmed that the transfer of charged solutes is the consequence of both size and ionic exclusion. Better performance was achieved with higher ionic strength (1 M).

The effects of heat treatment of ewes' milk at 65–85°C for 10–30 min and subsequent storage at 20°C for 90 min on the changes in ionic calcium and the distribution of calcium, magnesium and phosphorus between the diffusible and micellar phases were studied. As milk was heated there was a decrease in ionic calcium of ∼36 mg/l, but there was a rapid 86–90% recovery during the first 30 min after heating. Although the rate of reversal on subsequent storage decreased with the intensity of the heat treatment, there were no significant differences between samples after 90 min storage. Similar results were obtained for cows' milk subjected to the same treatments. Diffusible calcium, magnesium and phosphorus in heated samples were lower than in raw milk. Recoveries of initial values decreased with the severity of the heat treatment and were 80–95% for calcium, 85–97% for magnesium and 87–98% for phosphorus. Heat treatment under milder conditions (85°C for 20 s) was associated with almost complete recoveries. Recoveries of the minerals studied were slightly lower in ewes' than in cows' milk.

A fraction derived from the membrane of milk fat globules (MFGM) was isolated from fresh raw cream. The adsorption of MFGM at the interface of oil-in-water emulsions was studied as well as the stability of the emulsions under different conditions (pH, temperature treatment) and the surface potential. The emulsions prepared with MFGM isolates were stable at neutral pH and were destabilized at low pH. By analysis of the protein adsorbed at the interface by polyacrylamide gel electrophoresis, we also determined that the MFGM that covered the surface of the oil-in-water emulsion could not be displaced by surfactants such as Tweens and Triton X-100 and was not affected by the presence of other proteins such as caseins and β-lactoglobulin added to the emulsion. A strong molecular interaction between the adsorbed MFGM, composed of phospholipid and protein, may exist at the interface, rendering the newly formed membrane very little affected by the presence of other surfactants. This MFGM-stabilized emulsion was characterized by a behaviour very different from that of emulsions stabilized by other milk proteins.

There is considerable interest in the possibility that diet-derived isoflavonoids may help in protection against a number of chronic diseases common in Western society. Based on animal studies, however, concerns have been raised that consumption of isoflavonoids by infants and young children may be undesirable. Clover contains isoflavonoids and therefore may represent, via milk, a source of isoflavonoids in the human diet. In this study the concentrations of daidzein (7, 4′-dihydroxyisoflavone), genistein (5, 7, 4′-trihydroxyisoflavone) and equol (7-hydroxy-(4′-hydroxyphenyl)chroman) were measured using HPLC in cows' milk samples obtained from 76 farms in three Australian states. In addition, concentrations were measured in samples collected from one South Australian factory both before and after pasteurization. Concentrations in all samples were found to be extremely low. The mean daidzein concentration was <5 ng/ml. Mean genistein concentrations ranged from just detectable (∼2 ng/ml) in Victorian samples collected during summer to 20-30 ng/ml in samples from all states collected during spring when isoflavonoid-containing clover is most dominant in pasture. Mean equol concentrations ranged from 45±10 ng/ml in Victorian farm samples collected during summer to 293±52 ng/ml in Western Australian samples collected in spring. The mean concentrations of genistein and equol in post-pasteurization samples collected in spring were approximately double those for samples collected in autumn. Pasteurization had no effect on isoflavonoid concentrations. We conclude that the concentrations of isoflavonoids in Australian cows' milk are low and are therefore unlikely to have any pronounced biological effects in human consumers.

To estimate the suitability of cultured milk for the subsequent growth of dairy lactic acid bacteria in cheese manufacturing, control milk was first cultured until the end of the exponential phase with one of four proteolytic (Prt+) strains of Lactococcus lactis differing in their proteolytic enzymes, and pasteurized after readjusting the pH to its initial value. Nineteen non-proteolytic (Prt−) strains of Lc. lactis and nine Leuconostoc mesenteroides strains of dairy origin were then grown in the precultured milk until the stationary phase and their growth was compared with that in control milk. Despite the accumulation of non-protein N (NPN) during preculture, the growth of most Prt− strains of Lc. lactis in precultured milk was either reduced or unchanged whereas that of Ln. mesenteroides strains was unchanged or slightly stimulated. This reduction in growth was reversed by adding an NPN source to precultured milk, indicating that it was due to the exhaustion of assimilable NPN in precultured milk. Thus, preculturing milk with Prt+ strains of Lc. lactis could not be recommended for promoting the subsequent growth of starter cultures in cheese manufacturing.

Citrate metabolism by resting cells of Propionibacterium freudenreichii subsp. shermanii was investigated. In vivo13C nuclear magnetic resonance spectroscopy was used to study the pathway of citrate breakdown and to probe its utilization, non-invasively, in living cell suspensions. [2,4-13C]citrate was metabolized by resting cells to glutamate labelled in positions 2 and 4. In the presence of lactate or pyruvate, its rate of consumption was faster, but it was still converted to glutamate. No catabolic pathway other than the first third of a turn of the tricarboxylic acid cycle was used by Prop. freudenreichii subsp. shermanii to degrade citrate.

Polyclonal antibodies specifically directed towards native casein fractions have been recently used to solve some analytical problems such as localization of casein antigenic sites (Otani et al. 1985; Ametani et al. 1987), identification of casein variants (Moio et al. 1989a; Chianese et al. 1992), detection of bovine casein adulterating goats', ewes' and water-buffalo milk and cheese (Elbertzhagen & Wenzel, 1982; Bernhauer et al. 1983; Aranda et al. 1988; Moio et al. 1992; Rolland et al. 1993, 1995; Addeo et al. 1995b), evaluation of the efficiency of chromatographic fractionation of casein (Addeo et al. 1992) and detection of casein proteolysis in cheese (Addeo et al. 1995a). However, the use of polyclonal antibodies for immune analysis is often limited owing to nonspecific binding. The use of monoclonal antibodies may overcome this problem to some extent. Although the binding affinity of a hyperimmune serum is seldom attainable with a monoclonal antibody, an alternative approach consists of the isolation of specific antibodies by affinity chromatography (Johnstone & Thorpe, 1982). In a single step 1000–10000-fold purification has been achieved.

In this paper a fast protein liquid chromatography (FPLC) tandem immunoaffinity is used to enhance the specificity of bovine αs1-casein (CN) polyclonal antiserum. In the first column a caprine whole casein lacking αs1-CN was bound to a solid phase matrix and in the second column bovine αs1-CN was bound. After elution of macromolecular contaminants by a washing step, the two columns were detached and the purified bovine αs1-CN antibodies were eluted from the second column by a simple pH change.

In Italy buffalo milk is an important animal product utilized solely for the manufacture of Mozzarella cheese. Of an estimated population of 200000 buffalo there are ∼25000 controlled animals. The average milk production, expressed over 270 d lactation, is 2000 kg/head with average fat and protein contents of 82·6 and 46·4 g/l respectively (Associazione Italiana Allevatori, 1996). In recent years there has been a steady increase in the number of dairy buffalo replacing dairy cows as a consequence of the European Union quota system.

The cheesemaking qualities of milk depend on many factors, the most important of which are the concentrations of intact casein and fat. Milk in which casein has been broken down by proteolytic enzymes is of less value to cheese manufacturers (Lucey & Kelly, 1994). Plasmin (EC 3.4.21.7), the most important endogenous milk proteinase, occurs in milk together with its inactive proenzyme, plasminogen (Schaar & Funke, 1986). Plasmin hydrolyses αs-casein and β-casein, although κ-casein has been reported to be resistant (Fox, 1981). However, Andrews & Alichanidis (1983) found κ-casein to be hydrolysed quite rapidly by plasmin. Plasmin activity is higher in mastitic than normal milk (Bastian & Brown, 1996). Stage of lactation affects plasmin activity: late lactation is associated with higher concentrations of plasmin (Gilmore et al. 1995; Baldi et al. 1996). Thus, plasmin could be a major problem in herds with seasonal breeding such as buffalo, which progress through lactation in synchrony and are therefore at a similar stage of lactation at a given time.

The cascade of reactions leading to plasminogen activation is regulated by a complex network of molecular interactions between plasminogen activators (PA; EC 3.4.21.31) and at least three types of specific PA inhibitors (PAI-1; PAI-2; PAI-3; Saksela, 1985; Politis, 1996). There are two types of highly specific PA: tissue-PA (t-PA) and urokinase-PA (u-PA) (Saksela & Rifkin, 1988). PA activity is higher in mastitic than normal milk (Heegaard et al. 1994a). However, previous studies on PA in milk were carried out on bovine and caprine milk. No information is available on the presence and type of PA in buffalo milk. Thus the objective of the present study was to determine the level and type of PA in different fractions of buffalo milk: casein, serum, and somatic cells.