Macrophages isolated from the involuted bovine mammary gland were cultured in vitro. Phagocytosis of opsonized Staphylococcus aureus occurred rapidly, but intracellular killing of bacteria was slow. Many intracellular staphylococci survived for up to 4 d exposure to extracellular cloxacillin and emerged from within the macrophages to multiply extracellularly when the antibiotic was inactivated. Rifampicin was significantly more efficient than cloxacillin in killing intracellular S. aureus after 18 h incubation, but it too failed to sterilize the cultures within 3 d. Staphylococci, which had remained viable within macrophages during 20 h incubation with extracellular cloxacillin, showed an increased sensitivity to dilute lysostaphin on subsequent exposure. A 3 d course of intramammary therapy with cloxacillin, commencing simultaneously with an infecting inoculum of ∼108 colony forming units (c.f.u.) S. aureus, apparently eliminated the infection from one quarter of the udders of each of three lactating cows, but bacteria were re-isolated from two cows after a delay of several days. However, when other quarters of the same cows were infected with ∼108 c.f.u. S. aureus which had been phagocytosed by autologous mammary macrophages, similar simultaneous antibiotic therapy failed to affect these infections. The in vitro and in vivo findings indicate the significance of intracellular survival of S. aureus as a factor contributing to failure of antibiotic therapy.

1. A diffusion cup technique using Micrococcus lysodeikticus has been found suitable for lysozyme estimation. By this method raw milk gave only small zones of inhibition. The inhibitory substance of milk was shown not to be lysozyme.

2. Dose-response curves (dilution v. pH) were obtained with Streptococcus pyogenes as the test organism. They indicated that the effect of milk on this organism was the result of the interaction of one stimulatory and at least two inhibitory substances.

3. The existence of two inhibitory substances was confirmed by dilution assays, one, L1, occurring mainly in colostrum, the other, L2, occurring mainly in milk. The inhibition is greatest when L1 and L2 act simultaneously.

4. The pH and heat stabilities of the two substances are slightly different. L1 is most stable at pH 6−6·5, 90% being destroyed at 68° C. L2 is most stable at pH 7 and 95% is destroyed at 74°C.

Variations in mammary glucose uptake were measured during the normal pregnancy-lactation cycle in dairy goats. In addition mammary glucose uptake was studied in response to somatotropin (ST) treatment in mid-lactation and acute increases in glucose concentration induced by sodium-propionate challenge in early lactation. Mammary glucose uptake was independent of arterial glucose, insulin and Insulin-like Growth Factor-1 (IGF-1) concentrations during lactation and during acute increases in arterial glucose concentration. Glucose uptake in the lactating mammary gland of the goat must therefore be carried out by an insulin-independent carrier, possible GLUT1, and glucose supply is not a limiting factor for uptake under in vivo conditions. Extraction of glucose uptake changed markedly during the normal course of lactation, following the overall changes in milk yield. Concentrations of glucose in skimmed milk, believed to reflect intracellular glucose concentration, changed in opposite directions, resulting in decreasing ratios of arterialratioskimmed milk glucose concentration with progressing lactation. Thus, mammary synthetic capacity also involves a capacity for glucose uptake, which may be influenced by variations in glucose carrier numbers, as well as mammary metabolic activity (intracellular glucose concentration). In contrast to the situation during the normal course of lactation, ST stimulated milk yield, despite less efficient glucose extraction.

As moderate dilution causes little change in the surface tension of milk, it is shown to be advantageous to use σ-dilution curves in place of the σ-values of the undiluted fluid as a characteristic of the surface properties of milk. The complications arising from the presence of the milk fat are described, and it is suggested that the influence of the fat is of a physical rather than of a chemical nature. A study of the role of the various milk proteins shows the casein to be of great importance, whilst the heat-coagulable proteins have little influence. The serum obtained after removal of the casein and heat-coagulable proteins contains a residual fraction of protein-like material which is markedly surface active though constituting only about 3% of the total milk proteins. The surface-active material (σ-proteose) has been concentrated and isolated, and its properties are described in some detail.

Similar, marked variations in the freezing-point depressions of jugularvenous blood and of milk throughout a day were observed in cows when drinkingwater was offered for a single, short period each day. Values for milk were found to agree more closely, however, with those for mammary-venous blood than with those for jugular-venous blood. It appears that milk is in osmotic equilibrium with the blood flowing through the udder continuously throughout the period the milk remains within the udder and not only during its formation, and that milk secretion causes a slight alteration in the osmotic pressure of fluids within the immediate locality of the mammary gland. Changes in the milk composition that occurred in association with the observed changes in freezing-point depression were consistent with a movement of water into or out of the udder in response to any change in the osmotic pressure of blood.

It is fairly easy to control the enzymic hydrolysis of proteins in alkaline conditions by measuring the base consumption required to keep the pH constant in the reactor. Unfortunately, however, base consumption is not related in any simple way to the degree of hydrolysis reached at any given moment and to establish this relationship it is essential to find out the mean pK of the α-amino groups released during the hydrolytic process. We have shown here that the correct mean pK value varies according to the pH of the working conditions and that the relationship between these values may depend upon the kind of protein and protease used. We have put forward a method for determining this relationship experimentally by using a given protein–protease system, consisting of an alkaline titration of the raw protein and when partially hydrolysed. We have tested the results predicted by our theoretical model by applying it to the hydrolysis of whey proteins with a bacterial protease from Bacillus licheniformis at 50 °C, pH 8·0. This model can easily be applied to any hydrolytic process involving the appearance of functional groups that are partially protonizable under the working conditions in question in order to follow the kinetics of the reaction via the consumption of the neutralizing agent required to keep pH constant.

The overall objective of this study was to investigate how relevant research publications address the validity of diagnostic methods for acute puerperal metritis (APM) in dairy cows, a disease commonly treated with antibiotic drugs. Therefore, a literature search was conducted in Journal of Dairy Science, Theriogenology, Animal Reproduction Science and The Veterinary Journal utilizing the ScienceDirect database. The search revealed 259 articles addressing APM. After applying exclusion criteria, a total of 48 trials remained. It was determined whether the author gave a clear definition of APM, the time of diagnosis relative to calving, and the person who performed the diagnosis. Studies were checked for the presence of definitions of possible findings, thresholds, and test characteristics of the methods used. Overall 9 different diagnostic methods were employed. On average 2·5 ± 1·75 diagnostic methods were used in a study. References to support the use of the diagnostic methods were provided in 10 of 48 articles (20·8%). Vaginal discharge, transrectal palpation and rectal temperature were examined in 39, 22, and 21 of the studies, respectively. Thresholds for diagnostic tests and test characteristics were mentioned in 6 and 3 of the 48 articles, respectively. Based on this systematic review of 48 research papers the evidence supporting the use of the diagnostic methods to identify cows with APM has either been not reported or is weak. In conclusion, the reporting of the diagnostic methods to identify cows with APM needs to be improved and further high-quality research is necessary to improve diagnostic performance of the methods employed.

The aim of this study was to determine the influence of the somatic cell content (SCC) of milk on Parmigiano-Reggiano cheese yield, produced in commercial cheese factories under field conditions. The study was carried out following the production of 56 batches of Parmigiano-Reggiano in 13 commercial cheese factories by processing milk collected from Italian Friesian cattle herds. The vat-milk (V-milk) used for making each cheese batch was obtained by mixing evening milk (partially skimmed following spontaneous separation of fat overnight, natural creaming) and morning milk. The batches of cheese produced were divided into 5 classes according to the SCC value of the evening milk determined prior to natural creaming (class 1, from 0 to 200 000; 2, 201 000–300 000; 3, 301 000–400 000; 4, 401 000–500 000; 5, over 501 000 cells/ml). The cheese yield was calculated as the amount of 24-h cheese, expressed in kilograms, obtained from 100 kg of V-milk (24 h ACY). The values of fat, crude protein, true protein, casein and 24 h ACY of V-milk were negatively correlated with the somatic cell score (SCS) of the evening milk. Conversely, a positive correlation was observed between chloride and SCS. Fat, protein fractions (crude protein, casein and whey proteins), P and titratable acidity of V-milk were positively correlated with its 24 h ACY, while chloride, pH and SCS showed a negative correlation. A significant drop in 24 h ACY was observed in classes 3, 4 and 5, therefore when the SCC of the evening milk exceeded 300 000 cells/ml. Finally a lower recovery of milk fat in cheese was observed as SCC of evening milk increase.

Three groups of coryneform bacteria isolated from the rind of nine soft and semi-hard cheese varieties, i.e. the orange, white and yellow strains, were studied with regard to morphological and physiological features. The 115 orange isolates of the Brevibacterium linens type formed only one homogeneous phenon. The 96 white and 161 yellow pigmented strains were grouped into 15 phena. Five phena (58% of the strains) belong to the species ‘B. ammoniagenes’, Arthrobacter variabilis and A. nicotianae. The other strains were arranged into five Arthrobacter spp. and four Corynebacterium spp. groups as well as a Rhodococcus sp. group. Usually specific forms for the individual cheese varieties were involved. In cheeses of poor surface-smear quality B. linens was found in reduced numbers.

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.

Forty-eight Manchega dairy ewes were used during a complete lactation in a 2×2 factorial design to determine the effects of supplementing diets with fat (calcium soaps of palm oil fatty acids, CSFA) and rumen undegradable protein (RUP) on milk production and composition. Factors tested were amounts of CSFA (0 or 200 g/kg) and RUP (300 or 450 g/kg crude protein) in the concentrate. RUP was altered by adding a mixture of maize gluten meal and blood meal. Lactation was divided into one nursing period (period 1, weeks 1–4), and three milking periods (periods 2–4, weeks 5–8, 9–14 and 15–21). Concentrates were given at 0–8 kg/d during periods 1 and 2, and at 0–6 kg/d in periods 3 and 4. Ewes grazed rotationally in an Italian rye-grass pasture and received a daily supplement of 0·8 kg vetch–oat hay during period 1, and 0·3 kg lucerne hay during periods 2–4. For the whole lactation, supplemental fat markedly increased milk fat content (+23%) and yield (+16%), and decreased milk protein content (−9%). The positive effect of feeding CSFA on milk fat content was more evident at the beginning of lactation; however, its negative effect on milk protein was more pronounced in late lactation. Supplementary RUP had little effect, increasing milk protein content only in period 3, when the crude protein content of pasture was lower. Milk yield and lamb growth were not affected by dietary treatments. The results indicated that CSFA can be useful for increasing the milk fat content of dairy ewes at pasture, which may help farmers to produce milk reaching the minimum requirements of fat content for the cheese industry.

The changes in milk fat globules and fat globule surface proteins during concentration of whole milk using a pilot-scale multiple-effect evaporator were examined. The effects of heat treatment of milk at 95 °C for 20 s, prior to evaporation, on fat globule size and the milk fat globule membrane (MFGM) proteins were also determined. In both non-preheated and preheated whole milk, the size of milk fat globules decreased while the amount of total surface proteins at the fat globules increased as the milk passed through each effect of the evaporator. In non-preheated samples, the amount of caseins at the surface of fat globules increased markedly during evaporation with a relatively small increase in whey proteins. In preheated samples, both caseins and whey proteins were observed at the surface of fat globules and the amounts of these proteins increased during subsequent steps of evaporation. The major original MFGM proteins, xanthine oxidase, butyrophilin, PAS 6 and PAS 7, did not change during evaporation, however, PAS 6 and PAS 7 decreased during preheating. These results indicate that the proteins from the skim milk were adsorbed onto the fat globule surface when the milk fat globules were disrupted during evaporation.

The effects on rumen fermentation and milk-fat secretion of a dietary supplement of protected tallow given to 4 Friesian cows established on a low-roughage ration and with depressed milk fat is reported. The ratios of acetate to propionate in the rumen were unaffected by the supplement and remained typical of those associated with low-roughage diets in that the proportion of propionate was increased. The supplement produced almost complete recoveries in yield and content of milk fat without any increase in intramammary fatty-acid synthesis. The recoveries were due to transfer of about 20% of the total fatty acids of the tallow supplement. These results are discussed in relation to the effects of low-roughage diets on milk-fat secretion and it is concluded that in the ‘low-fat syndrome’ the capacity of the mammary gland to absorb preformed fatty acids is not impaired.

The concentrations of lactose, potassium, sodium, chloride and citric acid were determined in milks of cows, sows, ewes and goats. For cows, sows and ewes trends with lactation are reported and for all species correlations between constituents in milks secreted in mid-lactation are given. The results are discussed in relation to the secretion of water-soluble constituents in milk.

At atmospheric pressure, inactivation of lactoperoxidase (LPO) in milk and whey was studied in a temperature range of 69–73 °C and followed first order kinetics. Temperature dependence of the first order inactivation rate constants could be accurately described by the Arrhenius equation, with an activation energy of 635·3±70·7 kJ/mol for raw bovine milk and 736·9±40·9 kJ/mol for diluted whey, indicating a very high temperature sensitivity. On the other hand, LPO is very pressure resistant and not or only slightly affected by treatment at pressure up to 700 MPa combined with temperatures between 20 and 65 °C. Both for thermal and pressure treatment, stability of LPO was higher in milk than in diluted whey. Besides, a very pronounced antagonistic effect between high temperature and pressure was observed, i.e. at 73 °C, a temperature where thermal inactivation at atmospheric pressure occurs rapidly, application of pressure up to 700 MPa exerted a protective effect. At atmospheric pressure, LPO in diluted whey was optimally active at a temperature of about 50 °C. At all temperatures studied (20–60 °C), LPO remained active during pressure treatment up to 300 MPa, although the activity was significantly reduced at pressures higher than 100 MPa. The optimal temperature was found to shift to lower values (30–40 °C) with increasing pressure.

Cows managed for extended lactations of 16 months duration were milked on a half-udder basis twice or thrice daily, commencing in lactation week 9. Mammary epithelial integrity (assessed by milk sodium[ratio ]potassium ratio) was greater in the half-udder which was milked thrice daily. This difference was evident throughout the lactation but became greater after week 41. Milk protein composition was assessed during late lactation (52±3 weeks). Casein number (casein as a proportion of total protein) was significantly higher in half-udders milked thrice daily, as were the relative amounts of α- and β-caseins, whilst those of κ- and γ-caseins were reduced. Two days of inverted milking frequency (i.e. thrice-milked udder halves now milked twice, and vice versa) only partly reversed these differences. We concluded that thrice-daily milking will help to prevent or ameliorate the usual decline in milk processing quality associated with late lactation. Part of this effect is due simply to reduced exposure to proteolytic enzymes as a result of decreased storage time in the udder, but part is due to a better maintenance of epithelial tight junction integrity as lactation advances, which restricts leakage of proteolytic enzymes from serum into milk.

Multiple regression analysis has been used to relate contributions of individual flavour components to the flavour of Cheddar cheese. Equations have been derived that relate Cheddar flavour to the headspace concentrations of 2-pentanone, methanethiol and methanol. The concentrations of ethanol and H2S found in cheese headspace have been related to the flavour defects ‘fruity’ and ‘sulphide’ respectively

Unsupplemented acid goat whey containing 0·96% protein and 2·76% lactose was fermented aerobically with 32 microflora extracted from various raw milk cheeses and dairy products. These microflora were screened for their ability to hydrolyse whey proteins (α-lactalbumin and/or β-lactoglobulin) and to generate peptides inhibitors of Angiotensin I Converting Enzyme. Five microflora were able to degrade whey protein. The most efficient microflora was able to fully hydrolyse α-lactalbumin and to a lesser extend β-lactoglobulin. It was extracted from Bamalou des Pyrenées cheese. Micro-organisms involved consisted of yeast Kluyveromyces marxianus and lactobacillus Lactobacillus rhamnosus. Both were able to produce ACE inhibitory peptides after whey fermentation.

We describe a case of listerial mastitis in a flock of 130 sheep. The animals were housed at a farm where the bulk raw ewe milk was processed to produce raw milk soft cheese. List. monocytogenes was shed from the right mammary complex. Shedding was observed over a period of 99 d. A mean level of 4·56×104 cfu (colony forming units) Listeria monocytogenes/ml was recovered from the raw milk originating from the infected udder. The numbers ranged from 9×101 to 2·95×105. The bulk milk was contaminated by approx. 5·7×103 cfu/ml. In the cheese product, 2·0×102 cfu List. monocytogenes/g were constantly detectable for a period of 7 d post manufacture. The starter culture used for coagulation had a pivotal influence on the behaviour of List. monocytogenes during cheesemaking. Using the same mesophilic buttermilk culture as used by the farmer allowed numbers of Listeria to increase 60-fold within 12 h owing to a delayed acidification of the bulk milk. Addition of a thermophilic yogurt culture reduced the numbers of Listeria within 8 h of incubation.