We have investigated the responses of cow teats to machine milking in a study of relatively newly installed commercial milking parlours fitted with one of two types of milking cluster. The first was a common type with a large claw volume (> 200 ml), 15–16 mm i.d. long milk tube, 10 mm short pulse tube, cluster weight < 3·2 kg and used alternate pulsation. The second was a more traditional type with a 150 ml claw bowl volume, 13·5 mm i.d. long milk tube, 8 mm short pulse tube, cluster weight ∼ 3·5 kg and used simultaneous pulsation. We scored ∼ 50 cows in each of 20 herds, all within 60 s of cluster removal, for changes from the premilking teat condition: teat colour (creation of reddening or blueness), firmness, thickening at the base of the teat associated with the position of the liner mouthpiece, and whether the teat duct orifice was open. There were statistically significant differences in the proportion of cows displaying these four alterations in teat condition between herds using the two types of cluster. The more common type of cluster was always associated with better teat condition. The cause and effect of poorer teat condition have not been fully established and are likely to be multifactorial. The principal risk factors may be cluster weight, overmilking, vacuum applied during the overmilking phase and the design of the liner mouthpiece.

A series of experiments was conducted to investigate the effects of sodium fertilizers and supplements on the milk production and mammary health of dairy cows. In Expt 1, where sodium fertilizer was applied to productive pastures consisting mainly of the natrophile perennial rye-grass, the herbage sodium content and the milk yield of cows was increased and milk somatic cell count (SCC) reduced. In Expt 2, which used pastures containing less productive, natrophobic grasses and broad-leaved plants in Estonia, sodium fertilizer did not increase herbage sodium content and did not affect milk production or composition. In Expt 3 the sodium content of the diet of individually tethered cows was increased from 1 to either 6 or 11 g/kg dry matter (DM) by adding salt to their restricted feed allowance, and the cows' milk yield was increased by the high level of sodium supplement and milk SCC were reduced by both levels of sodium supplement. The calcium and magnesium status of cows was improved by the sodium supplement. In Expt 4 a low level of supplementary salt was included in the ration of tethered cows to increase the sodium content of the diet from 2 to 3·6 g/kg DM. No effects on milk yield or SCC were found, but the sodium supplement reduced Staphylococcus aureus contamination of the milk, but not the proportion of milk samples infected with Escherichia coli. It was concluded that the optimum dietary sodium concentration for maximum milk yield was greater than the published requirements, and that substantial increases in sodium intake above current requirements also reduced milk SCC.

The aim of the study was to examine the effect of morphine and naloxone pretreatment on cortisol and ACTH concentrations in response to machine milking in dairy cows. In the first part of the experiment, the effects of i.v. morphine doses (0, 21, 70 and 210 mg, one dose each day) 10 min before morning milking were studied in six Brown Swiss dairy cows. In the second part, four cows were treated 1 d after the control milking with 210 mg morphine at 10 min before milking and the next day with 210 mg naloxone at 15 min before milking followed by 210 mg morphine at 10 min before milking. In addition, four other cows were treated 1 d after the control milking with 210 mg naloxone at 10 min before milking. Pretreatment with morphine significantly suppressed the machine milking-induced increase of cortisol in blood plasma as compared with controls. Naloxone pretreatment overcame the inhibitory effect of morphine and elevated milking-induced cortisol concentrations. Naloxone administration alone significantly increased cortisol concentration resulting from milking as compared with controls. However, ACTH concentrations did not change in either control or treated animals, suggesting an ACTH-independent release mechanism for cortisol during milking. We conclude that the release of cortisol in response to machine milking seems to be modulated by endogenous opioids at the adrenal level and does not appear to be under the control of ACTH.

The effects of level of fish oil inclusion in the diet on grass silage intake, and milk yield and composition of dairy cows offered either 5 or 10 kg concentrates/d were evaluated in a ten treatment, partly balanced, changeover design experiment involving 50 cows in early lactation. Concentrates were prepared to provide 0, 150, 300 or 450 g fish oil/cow per d or 300 g fish oil/cow per d from a premix when each animal was offered 5 kg/d. The fish oil was predominantly from herring and mackerel caught in the North Atlantic while the fish oil premix was obtained from a commercial source and used palm kernel expeller as a carrier. Increasing fish oil supplementation decreased silage dry matter intake and the concentrations of milk fat and protein, and increased milk yield and diet digestibility. There were significant interactions between concentrate feed level and level of fish oil for silage intake and milk yield. Other than for the concentrations of milk fat and protein, and 20[ratio ]4n−6 fatty acids, the source of fish oil did not affect forage intake or animal performance. Fish oil supplementation also decreased the concentrations of milk protein by 0·9 g/kg for each 100 g increase in fish oil supplementation, the depression being similar at each level of concentrate feeding. Supplementing the feed of dairy cows with 450 g fish oil/cow per d decreased the concentration of milk fat by 15 g/kg. This study also showed that feeding dairy cattle with fish oil is an efficient method of increasing eicosapentaenoic acid in the human diet through transfer into milk.

The progress of Staphylococcus aureus infection from inoculation to the early chronic stage was examined in 12 Israeli-Holstein cows (four primiparous and eight multiparous) for up to 48 d after inoculation. Before inoculation, the primiparous cows were free from any infection and the multiparous cows were infected by coagulase-negative staphylococci. Two quarters in each cow were inoculated intracisternally following milking with 2000 cfu of a local prevailing Staph. aureus strain, VL-8407. Infection was established in 21 out of 24 quarters. The control quarters remained free from infection during the study, with no significant change in function. No statistically significant differences were found between primiparous and multiparous cows in the responses examined. Somatic cell count (SCC) increased within 24 h of inoculation and remained high for the duration of the study. In the infected quarters mean ln (SCC) increased within 24 h from 9·9±0·5 before inoculation to 13·0±0·2 after inoculation; most of the cells were neutrophils. N-acetyl-β-glucosaminidase activity, expressed as ln (nnmol/min per l), was increased from 0·9±0·6 to 2·4±0·2 by inoculation, and was highly correlated with SCC. The Staph. aureus count fluctuated with no particular relationship with SCC. The phagocytic activity of neutrophils was significantly lower in the inoculated than in the control quarters and this difference increased with time after inoculation. CD8+ T lymphocytes were the main subpopulation of lymphocytes found in inoculated quarters. After inoculation, maximum but not minimum electrical conductivity (EC) recorded during milking increased significantly. The rises in maximum EC varied significantly among cows. The rises in SCC were associated with a persistent increase in EC in only one of the eight cows examined. No clinical signs were observed, and milk yield and composition were not affected during the study period. The results suggest that some strains of Staph. aureus may induce a relatively mild response in mammary glands of cows in mid lactation, and that the concomitant development of such chronic Staph. aureus infections in two quarters may not be detected by changes in the EC of composite milk and in the yield of the cow.

The mechanism of glycine transport in lactating mouse mammary gland was investigated. Three Na+-dependent systems of glycine transport, distinguished on the basis of their ionic requirement and sensitivity to 2-(methylamino)isobutyric acid (MeAIB), were A (Na+-dependent, MeAIB-sensitive); (Na++Cl−)-dependent, MeAIB-insensitive; and Na+-dependent, Cl−-independent, MeAIB-insensitive. These systems were further distinguished on the basis of inhibition analysis and sensitivity to pH of the extracellular medium and preloading mammary tissue with amino acids. The uptake of glycine via the A system (Km 0·53 mM) was inhibited by preloading mammary tissue with alanine, while glycine uptake mediated by the (Na++Cl−)-dependent, MeAIB-insensitive system (Km 0·47 mM) was downregulated by preloading mammary tissue with all amino acids (alanine, sarcosine and histidine) tested. Treatment of mammary tissue with N-ethylmaleimide inhibited the uptake of glycine via both these systems. Decreasing the pH of the extracellular medium inhibited the uptake of glycine via the A system but not the (Na++Cl−)-dependent, MeAIB-insensitive system. On the basis of ionic requirement, system A appears to comprise two components, one dependent on Na+ plus Cl− and the other on Na+ alone. Insulin upregulated the A system-mediated uptake of glycine in pregnant mouse mammary tissue cultured in vitro, while the (Na++Cl−)-dependent, MeAIB-insensitive system remained unaffected.

The aim of this study was to investigate the action of opioids (the μ receptor agonist morphine) and the antagonist naloxone on inhibition of oxytocin release and milk let-down in response to milking in dairy cows. In the first experiment, cows were injected with 0, 21, 70 and 210 mg morphine 10 min before milking on four successive days. Plasma oxytocin levels after 1 min manual stimulation of the udder were reduced by 70 and 210 mg morphine, and milk let-down was inhibited at the latter dose. In the second experiment, cows were injected after a control milking with 210 mg morphine (or 350 mg at 10 min before milking the following day if not effective) to inhibit milk flow. On the following day the inhibiting dose of morphine was given with 210 mg naloxone. Naloxone injection given before morphine had no effect on plasma oxytocin concentrations, but abolished the inhibition of oxytocin release by morphine and potentiated oxytocin release in response to milking. Naloxone alone injected the day after control milking increased oxytocin levels during milking, suggesting involvement of the opioid system in milking. A model has been developed for the control of opioid effects during milking. Morphine suppressed oxytocin release during milking in a dose-dependent manner and the effect was reversible by naloxone.

The effect of tension of teatcup liners on teat end condition and quantity of keratin in the teat canal was investigated. Liner tension was increased by using longer teatcup shells. The first experiment used six Holstein cows in early lactation. Left quarters were milked with liners under medium or normal tension by using Conewango liners in 142 mm shells. Right quarters were milked with liners under high tension by mounting the liners in teatcup shells 149 mm in length. By day 16, teat end condition and sensitivity to manipulation were worsened by thrice daily milking when liners were under a higher tension. Two subsequent experiments each used 12 different Holstein cows. These cows were in mid lactation and were milked twice daily for 10 or 30 d. Left quarters were milked with liners under high tension. Right quarters were milked with liners under low tension by using teatcup shells 126 mm in length. The quantity of keratin removed during milking was not influenced by liner tension; however, the quantity of keratin at the end of the experiments was increased 10–20% in teats that were milked using liners under a higher tension. Histological analysis and keratin content were consistent with epithelial hyperplasia induced by milking with liners under increased tension.

The mechanism of cationic amino acid transport in lactating mouse mammary gland was investigated. Two Na+-independent systems of arginine transport were discriminated on the basis of their sensitivity to leucine. The leucine-sensitive uptake of arginine (Km 0·4 mM) was through a broad specificity system that interacted with both cationic and neutral amino acids, and was inhibited by preloading mammary tissue with neutral amino acids. The leucine-insensitive uptake was identified as the y+ system (Km 0·76 mM). Preloading mammary tissue with cationic amino acids increased the uptake of arginine by the y+ system. Decreasing the pH of the external medium to 6·0 suppressed the y+ system-mediated uptake by ∼25%, whereas the broad specificity system remained unaffected. Lactogenic hormones upregulated the y+ system-mediated uptake of arginine in pregnant mouse mammary tissue cultured in vitro, although the broad specificity system remained unaffected. The y+ system-mediated uptake increased 2-fold with insulin alone and 4-fold with the combination of insulin, cortisol and prolactin.

A kinetic modelling approach was developed and investigated with the aim of predicting the utilization of major substrates in the mammary gland and milk secretion rates in the lactating cow at varying concentrations of substrate in arterial blood. The model includes kinetic equations of transport and metabolism of glucose, acetate, free amino acids and free fatty acids in secretory cells and a phenomenological description of autoregulation of local blood flow, in which an energy criterion of control has been used. The predicted relationships between the rate of milk secretion and glucose levels in the blood are consistent with experimental results. Differential stimulation of α-lactalbumin synthesis causes increments in local blood flow and milk secretion rate in the model. The results of the study suggest that there is no simple relationship between the level of substrates in the blood and milk yield and contents of fat and protein in milk. This is because the effect on production of varying patterns of substrate concentrations in the blood is mediated by network interactions at the level of secretory cell metabolism and microcirculation. However, dynamic modelling provides a rational framework for developing such predictive tools.

We have studied the effect of treatment with enrofloxacin on local and general clinical signs and chemiluminescence of circulating polymorphonuclear leucocytes during experimentally induced Escherichia coli mastitis in cows immediately after parturition. Twelve cows were infected with 104 cfu Esch. coli P4[ratio ]O32 into both left quarters. Six cows received an intravenous injection of 5 mg enrofloxacin/kg at 10 h after infection and a second enrofloxacin treatment administered subcutaneously at 30 h post infection. The other six cows were controls that received no treatment. General clinical signs (fever, tachycardia, loss of appetite, reduced rumen motility and depression) were similar in both groups. Local clinical signs, such as swelling, pain and firmness of the inflamed mammary quarters, were less severe in the treated cows. We saw no difference in the appearance of the milk[ratio ]flecks and watery or purulent milk were observed in both groups. The beneficial effects of treatment with enrofloxacin were mainly on milk production and composition. The decline in milk production and the changes in milk concentrations of lactose, Na+ and bovine serum albumin were less pronounced in the treated cows. Treatment with enrofloxacin accelerated the clearance of bacteria from the infected quarters, but had no effect on the chemiluminescence response of isolated polymorphonuclear leucocytes. The changes in the number of circulating leucocytes and the appearance of immature neutrophils in the circulation of the treated cows indicated possible beneficial effects on migration of neutrophils into the inflamed glands. Higher milk somatic cell counts in the treated cows supported this hypothesis. The results of this study indicated that treating cows that have been experimentally infected with Esch. coli mastitis after parturition with enrofloxacin reduced the severity of the disease, especially the decline in milk production and the changes in milk composition.

Heat (85 °C for 20 min) and pressure (600 MPa for 15 min) treatments were applied to skim milk fortified by addition of whey protein concentrate. Both treatments caused > 90% denaturation of β-lactoglobulin. During heat treatment this denaturation took place in the presence of intact casein micelles; during pressure treatment it occurred while the micelles were in a highly dissociated state. As a result micelle structure and the distribution of β-lactoglobulin were different in the two milks. Electron microscopy and immunolabelling techniques were used to examine the milks after processing and during their transition to yogurt gels. The disruption of micelles by high pressure caused a significant change in the appearance of the milk which was quantified by measurement of the colour values L*, a* and b*. Heat treatment also affected these characteristics. Casein micelles are dynamic structures, influenced by changes to their environment. This was clearly demonstrated by the transition from the clusters of small irregularly shaped micelle fragments present in cold pressure-treated milk to round, separate and compact micelles formed on warming the milk to 43 °C. The effect of this transition was observed as significant changes in the colour indicators. During yogurt gel formation, further changes in micelle structure, occurring in both pressure and heat-treated samples, resulted in a convergence of colour values. However, the microstructure of the gels and their rheological properties were very different. Pressure-treated milk yogurt had a much higher storage modulus but yielded more readily to large deformation than the heated milk yogurt. These changes in micelle structure during processing and yogurt preparation are discussed in terms of a recently published micelle model.

High isostatic pressures up to 600 MPa were applied to samples of skim milk before addition of rennet and preparation of cheese curds. Electron microscopy revealed the structure of rennet gels produced from pressure-treated milks. These contained dense networks of fine strands, which were continuous over much bigger distances than in gels produced from untreated milk, where the strands were coarser with large interstitial spaces. Alterations in gel network structure gave rise to differences in rheology with much higher values for the storage moduli in the pressure-treated milk gels. The rate of gel formation and the water retention within the gel matrix were also affected by the processing of the milk. Casein micelles were disrupted by pressure and disruption appeared to be complete at treatments of 400 MPa and above. Whey proteins, particularly β-lactoglobulin, were progressively denatured as increasing pressure was applied, and the denatured β-lactoglobulin was incorporated into the rennet gels. Pressure-treated micelles were coagulated rapidly by rennet, but the presence of denatured β-lactoglobulin interfered with the secondary aggregation phase and reduced the overall rate of coagulation. Syneresis from the curds was significantly reduced following treatment of the milk at 600 MPa, probably owing to the effects of a finer gel network and increased inclusion of whey protein. Levels of syneresis were more similar to control samples when the milk was treated at 400 MPa or less.

Twelve cows were experimentally infected in two quarters with 1 × 104 cfu Escherichia coli per quarter and six cows were infused with 500 μg endotoxin into two quarters. Six cows infected intramammarily with Esch. coli were treated intravenously with a bactericidal antibiotic 10 h after infection and subcutaneously 20 h later. Blood and milk samples were collected from all cows at regular time intervals. Milk production decreased more rapidly, but was less pronounced, after endotoxin infusion than during Esch. coli mastitis. The milk production losses in the non-inflamed quarters were negligible in endotoxin mastitis, but were substantial during Esch. coli mastitis, probably due to more pronounced systemic effects. Reticulo-rumen motility was inhibited only during Esch. coli mastitis. Changes in plasma haptoglobin were more pronounced during Esch. coli mastitis, although they occurred sooner during endotoxin mastitis. No changes in plasma activities of enzymes such as lactate dehydrogenase, glutamic–oxaloacetic transaminase and γ-glutamyl transpeptidase were observed. Concentrations of tumour necrosis factor-α increased in both types of mastitis. Absorption of these cytokines into the circulation was highest during Esch. coli mastitis, especially in the untreated control group. We found only minor differences between the treated and untreated Esch. coli groups, but there were larger differences between the Esch. coli groups and the endotoxin group. These differences were probably due to differences in kinetics, composition and amounts of different cytokines released in the mammary gland and subsequently absorption into the circulation. Endotoxin is probably not directly responsible for the systemic changes during coliform mastitis.

Ovine casein macropeptide (CMP) was characterized by anion-exchange FPLC and reversed-phase (RP) HPLC. To study heterogeneity (the degree of glycosylation and phosphorylation), CMP was desialylated with neuraminidase and dephosphorylated with acid phosphatase. Following RP-HPLC, the main CMP components were identified using either on-line or off-line mass spectrometry. The most abundant ovine CMP component was a diphosphorylated carbohydrate-free form, followed by one or two monophosphorylated and a non-phosphorylated asialo-aglyco species. Aglyco non-phosphorylated, monophosphorylated and diphosphorylated forms were in the ratio 3[ratio ]20[ratio ]77. Only ∼ 30% of ovine CMP was glycosylated. Assuming that the monosaccharide fraction of ovine CMP is composed of N-acetylgalactosamine, galactose and N-glycolylneuraminic acid, molecular masses consistent with the presence of CMP containing tetra-, tri-, di- and monosaccharide were identified.

The physiological effects of intramammary infusions of recombinant bovine cytokines in six lactating dairy cows on the quality and yield of milk and the bactericidal activity of milk neutrophils were investigated. Recombinant bovine interleukin-2 (rboIL-2) and interferon-γ (rboIFN-γ) were produced in the yeast Pichia pastoris. Two animals were given rboIL-2 (2×105 units) in two quarters, two animals were given rboIFN-γ (6·5×105 units) in two quarters, and the other two cows received a dose of rboIL-2 in one quarter and rboIFN-γ in a second quarter. In addition, each animal was given phosphate-buffered saline (PBS) in the other two quarters as a control. Somatic cell counts and conductivity of the fore milk were monitored before and after infusion. Neutrophils were isolated from quarter milk samples 36 h after infusion of cytokine or PBS and their bactericidal activities against Staphylococcus aureus were measured in vitro with a colorimetric assay. Quarters infused with rboIL-2 or rboIFN-γ showed significant but transitory increases in both milk somatic cell counts and conductivity when compared with preinfusion values and with control quarters. There were minimal effects on daily milk yield. Neutrophils isolated from milk from quarters infused with rboIL-2 showed enhanced bactericidal activity against Staph. aureus. The bacterial killing from rboIL-2 treated quarters was significantly greater, with a mean of 63·5% compared with a mean of 5·4% for neutrophils taken from uninfected quarters to which PBS had been administered. The bactericidal activities for quarters treated with rboIFN-γ and infected quarters treated with PBS were 15·0 and 30·0% respectively. The results indicate that intramammary infusions of rboIL-2 and rboIFN-γ to lactating cows are well tolerated, and that rboIL-2 can activate milk neutrophils and augment their bactericidal activity.

This paper presents a novel contribution to the purification of goat β-lactoglobulin by using an ultrafiltration membrane enzymic reactor. The basis of the purification process was the enzymic hydrolysis of contaminating proteins, α-lactalbumin and traces of serum albumin, by pepsin at 40 °C and pH 2, conditions under which β-lactoglobulin is resistant to peptic digestion. Simultaneously, β-lactoglobulin and peptides were separated by ultrafiltration. β-Lactoglobulin was retained in the reactor while peptides generated by hydrolysis from α-lactalbumin and serum albumin permeated through the membrane. The process was made continuous by the addition of fresh whey to replace the lost permeate. Three mineral membranes with 10, 30 and 50 kDa molecular mass cut-off were tested and the 30 kDa membrane was selected for the continuous process. The simultaneous purification and concentration of β-lactoglobulin from clarified goats' whey was achieved in a single step. The ultrafiltration membrane enzymic reactor could treat eight reactor volumes of clarified whey. The recovery of β-lactoglobulin was 74%, its purity was 84% and its concentration 6·6-fold that in the initial clarified whey.

An adaptive stochastic dynamic programming model was used to solve the optimum replacement decision problem for the dairy cow under a range of alternative mastitis control procedures. The model predicted that reducing milk yield losses and somatic cell count penalties by using milking machine test, post-milking teat disinfection and dry cow therapy added approximately £4, £10 and £13 respectively to an original annuity equivalent net present value for the replacement heifer of £286. Assuming that these procedures also reduced involuntary culling due to mastitis by 50% added £8·90 to the annuity. This latter figure indicated that an important part of the benefit of mastitis control procedures might come from a reduction in the culling risk of persistent clinical cases. We concluded that the strength of the dynamic programming model in this context was that it provided an integrated evaluation of the various impacts of each alternative mastitis procedure in the long term, which is essential for correct economic evaluation of mastitis.

The effects of cold storage at 4 °C for 12, 24 and 48 h on the physicochemical characteristics and renneting properties of ewes', goats' and cows' milks were compared. The most important changes were observed in cows' milk. Soluble calcium concentrations were not affected in ewes' milk but were increased by 10% in cows' milk and 7% in goats' milk. More casein was dissociated on cooling cows' (+300%) than goats' (+100%) milk, and there was no change in soluble casein in ewes' milk. The coagulation characteristics of cows' milk were more impaired by cold storage than goats' or ewes' milks. Coagulation times increased by ∼30% and whey draining capacity decreased by 40% after cows' milk was cooled, but there were no changes with ewes' milk and only a slight decrease in coagulation time with goats' milk. We propose an interpretation of these results based on the physicochemical properties of each type of milk.

We have investigated the effects of adding a range of mineral salts and calcium-chelating agents on the distribution of casein and minerals between the non-pelleted and pelleted phases of milk obtained upon centrifugation at 78000 g for 90 min. Adding CaCl2 or mixtures of NaH2PO% and Na2HPO% to reconstituted skim milk (90 g milk solids/kg) at pH 6·65 increased both pelleted casein and pelleted calcium phosphate. Opposite effects were obtained by adding citrate or EDTA. The change in pelleted calcium phosphate was not simply related to casein release from the micelle. Upon adding 5 mmol EDTA/kg milk, 20% of the pelleted Ca, 22% of the pelleted phosphate and 5% of the micellar casein were removed. Increasing the concentration of EDTA to 10 mmol/kg milk decreased the pelleted Ca by 44% and the pelleted phosphate by 46%, and caused 30%of the micellar casein to be released. The effects of adding phosphate, citrate or EDTA at pH 6·65, followed by the addition of CaCl2, demonstrated the reversibility of the dissolution and formation of the micellar calcium phosphate. There were limits to this reversibility that were related to the amount of colloidal calcium phosphate removed from the casein micelles. Adding CaCl2 to milk containing [ges ] 20 mmol EDTA or [ges ] 30 mmol citrate/kg milk did not result in complete reformation of casein micelles. Light-scattering experiments confirmed that the dissolution of moderate amounts of colloidal calcium phosphate had little effect on micellar size and were reversible, while the dissolution of larger amounts of colloidal calcium phosphate resulted in large reductions in micellar size and was irreversible.