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.

The production of milk, fat and protein of 37216 mothers of twins was compared with that of 1691 127 mothers of single calves of Black and White breeds of cattle recorded in England and Wales. At every parity, the mothers of twins produced more milk, fat and protein than the mothers of single calves. The proportion of twin-bearing cows retained increased with parity, from 0·67% at first calving to ∼ 4·0% at the tenth or eleventh calving. More mothers of twins achieved 305 d of lactation subsequently than mothers of single calves, although their inter-calving intervals were not generally longer.

As mastitis is associated with leakage of small molecular weight plasma proteins, such as α1-antitrypsin, into milk, this antitrypsin can be used as an indicator of mastitis. A colorimetric procedure was developed for large scale monitoring of milk antitrypsin activity, using microtitration plates and the Multiskan system. The effect of stage of lactation and age of the cow on the antitrypsin concentration and its interrelationship with other mastitis indicators (bovine serum albumin (BSA), somatic cell count) was analysed by computer programs on 1029 cows. Milk antitrypsin activity was high after parturition owing to colostral inhibitors. After the first month of lactation the assay measures only blood-derived antitrypsin and is a good indicator for detecting an increased permeability between blood and milk due to mastitis. Increasing lactation number only slightly affected the antitrypsin and BSA concentrations whereas somatic cell content was markedly affected.

Housed dairy heifers (62 cows) were fed concentrates exclusively through variable-time, out-of-parlour dispensers. They were observed to visit the dispensers on average 20·4 times/d but significantly less frequently (13·9 times/d) shortly after calving. The dispensers were occupied for 25·8% of the night (22.00–06.00 h) compared with 30·4% of the day (06.00–22.00 h). This difference was small but significant, whereas the differences between the proportion of night and day spent standing (32·5 v. 66·2%) or eating silage (4·8 v. 24·7%) were noticeably greater. In a herd of ∼ 80 cows in which the daily average number of visits to the dispenser was 20·5/cow there was a significant decrease in the number of visits/d (y) with increasing time after calving (x) where y = 23·4–0·030x (r = 0·88). Some cows repeatedly visited the dispensers more frequently than others (range 13·2–34·9±5·3 visits/d, giving a repeatability of 0·58). Cows in a grazing herd (∼ 60) visited the dispensers on average 14·6 times/d during an average 3·6 visits to a cubicle house to which they had continuous access. The number of visits to the dispensers (y) again decreased with increasing time after calving (x) where y = 22·7–0·072x (r = 0·45). The average time spent in the cubicle house (z) also decreased as the days after calving (x) increased (z = 8·4 – 0073x+0·000203x2, r =–0·66). All cows, however, stayed in the cubicle house during periods of heavy rainfall.

Changes in the level of the tissue damage marker enzyme, N-acetyl-β-D-glucosaminidase (NAGase) in quarter fore milks were found to be related to the presence and types of pathogenic bacteria present and to somatic cell counts (SCC). Minor pathogens (coagulase-negative staphylococci, Corynebacterium bovis) elicited a mild SCC increase (from a mean of 243 × 103/ml in healthy quarters to 504 × 103/ml in infected quarters) with marginal tissue damage (mean NAGase activity increased from 21 in healthy quarters to 28 in infected quarters). Major pathogens (i.e. Staphylococcus aureus, Streptococcus agalactiae, Str. dysgalactiae and Str. uberis) caused more severe tissue damage (mean NAGase of 48) and SCC increases (mean, 2803 × 103/ml). The NAGase test could also be used effectively on composite milk samples where regular monthly NAGase analysis was able to identify correctly 74% of animals having infected quarters. The possibility of combining SCC and NAGase data in order to give a more definite diagnosis of bovine mastitis is discussed.

First lactation records of a total of 242 Jersey (JH), Brown Swiss (BH) and Holstein-Friesian (FH) halfbreeds from Hariana cows were studied for projecting 300 d milk yield from 12,14,16,18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38 and 40 weeks’ milk yield. The correlation coefficients between 300 d milk yield and part records ranged from 0·84 to 0·9 in the JH, 0·80 to 0·90 in the BH and 0·72 to 0·86 in the FH groups. Prediction equations and standard errors of estimation for prediction of 300 d milk yield for an individual animal are presented. It is suggested that part records up to 16, 22 and 22 weeks may be used for projecting 300 d yield with sufficient accuracy in the JH, BH and FH groups respectively.

Data from experiments in 55 commercial herds have been examined to study the patterns of Corynebacterium bovis infection. Such infections are most common in herds that are not subjected to regular control methods but their commercial significance appears to be small. In herds using teat disinfection and dry cow therapy levels of infection with this organism are low. There is no evidence from these data to suggest that C. bovis infections protect the udder against invasion by a major pathogen. On the contrary these infections do not persist when major pathogens invade.

Experiments are described in which lactating cows were exposed to Corynebacterium bovis either by dipping the teats in a suspension of the bacteria or by inoculating the bacteria into the teat duct or the teat sinus. All three methods readily led to ‘infection’ being established. The effect of these infections on somatic cell count was minor and no clinical mastitis resulted during the course of the experiments although some cases occurred subsequently. There was evidence that 44% of these infections were confined to the teat duct. Quarters excreting C. bovis in milk continued to do so during endotoxin-induced inflammation and showed a similar cellular response to that of uninfected quarters.

Quantitative cytological analysis demonstrated a marked and progressive increase in concentration of subepithelial plasma cells from milk-secreting parenchyma to the distal teat end mucosa in the normal, lactating bovine mammary gland. Parenchymal plasma cells exhibited typical ultrastructure with abundant, flattened stacks of rough endoplasmic reticulum, and hypertrophied Golgi components, while many cells in the teat end mucosa displayed an ergastoplasm distended with fine, flocculent material. Intraepithelial plasma cells were also observed in teat end mucosa. Results suggest that teat end tissues function by recruiting plasma cells to serve as protection against invading mastitis-causing organisms.

Effects of genetic variants of the milk proteins, αs1-casein, β-casein, κ-casein and β-lactoglobulin (βlg), on milk yield and composition, particularly the protein composition, were investigated in milk samples from 289 Jersey and 249 Friesian cows in eight commercial herds.

Milk protein genotypes had no significant effect on yields over a complete lactation of milk and fat, but significant differences in fat content were detected for β-casein (B, A1B, A2 > A1A2) and βlg (B, AB > A) variants. Significant differences between β-lg variants were also found with total solids (B, AB > A), casein (B, AB > A), whey protein (A > AB > B) and βlg (A > AB, AC > B > BC) concentrations. Casein genotypes were not significantly different in total protein and casein concentrations but many differences were found in casein composition. αs1-Casein variants significantly affected αs1-casein (BC > B) and κ-casein (B > BC) concentrations. β-Casein variants affected concentration and proportion of β-casein (A1B, A2B > A1, A1A2, A2, B), αs1-casein (A1, A2 > B) and κ-casein (B > A2) and concentration of whey protein (A1 > most other β-casein variants). κ-Casein variants affected concentration and proportion of κ-casein (B > AB > A), proportion of αs1-casein (A > AB > B) and concentration of βlg (A > AB, B) and α-lactalbumin (A, AB > B). Differences in milk composition were found between breeds, herds and ages, and with stage of lactation. The potential use of milk protein genotypes as an aid in dairy cattle breeding is discussed.

Growth of Streptococcus thermophilus was faster in mastitic milk, aseptically collected following intramammary infusion of Escherichia coli endotoxin, than in normal aseptically collected milk. The contribution of polymorphonuclear leucocytes (PMN), of plasma and plasmin to this stimulation has been investigated. Addition of plasmin to low cell count milk stimulated growth of the streptococcus to the same degree as mastitic milk. However, addition of plasma or PMN from blood or from milk were less stimulatory. The part played by these components in providing casein breakdown products for the weakly proteolytic Str. thermophilus in mastitic milk is discussed.

Oxytocin was infused for 60 min into the jugular vein of 8 lactating cows with full udders before milking. Under these circumstances milk loss occurred spontaneously in 3 cows and was evoked in 5 cows by reducing pressure around one teat (–30 cm water) using a plethysmographic chamber. Milk loss was measured continuously. The postsynaptic α-blocking agent prazosin (PRZS) injected either into the udder artery (2 or 5 mg) or into the jugular vein (200 mg) consistently induced a significant increase in milk leakage. It is suggested that PRZS effectively inhibits α-adrenoceptors in the teat sphincter muscles and thus promotes the action of β-adrenoceptors, in this way inducing relaxation of the teat sphincter.

In addition to a control diet, lactating cows were offered saturated fatty acid mixtures in three forms, free acids, free triglycerides and protected triglycerides, i.e. triglyceride encapsulated within a protein matrix which was cross linked by exposure to formaldehyde. Relative to the control diet, all three supplements increased milk yield. However, only the free fatty acids gave rise to increased yields of the three major milk components. The free fat and the protected fat caused significant increases only in the lactose yield. The different effects of the supplements on the yield of milk fat are suggested to be due to the types of long chain acid reaching the mammary gland rather than to any change in rumen activity. Changes in the concentrations of the soluble multivalent ionic constituents of the milks were consistent with this conclusion.

A practical effective method was devised for the removal of bacteria from soiled glass, rubber and stainless steel pipe sections which allowed the isolation of bacteria more loosely associated with the surfaces. This method, consisting of a pre-rinse followed by six successive double squeegee rinses, was compared with six other procedures (involving scraping followed by one or more successive swabbings) for the removal of bacteria from rubber pipe sections which had been inserted into an experimental milking plant. The squeegee method was found to be the most effective. It was also found that one particular part of these pipe sections was consistently more heavily contaminated.

Milk composition and yield were measured in the black Bedouin goat during 4 d of dehydration followed by 2 d of rehydration. During this time the goats were exposed to strong solar radiation with no access to shade; all of these conditions occur commonly in the desert. Milk yield, initially similar to that found in the desert, was maintained during the first 2 d of dehydration, fell to 35% of the initial value over the third and fourth days and recovered fully during 2 d of rehydration. As yield fell, milk osmolality, and milk fat and protein concentrations rose. During rehydration all 3 fell once more, but whereas osmolality finished below initial values the concentrations of fat and protein remained higher than before dehydration. The milk remained isosmolar with plasma throughout. Total yields of milk solids and milk water during the whole experimental period were 70 and 67% of normal respectively and normal growth of the young was not disturbed.

The effects of κ-casein genetic variants, lactation number, pH and casein content on the renneting properties of individual milks were studied. Rennet coagulation time (RCT) and curd firmness were determined with the aid of a lactodynamograph and the results statistically analysed. Renneting properties were strongly related to κ-casein variants and lactation number, with κ-casein B and higher lactation numbers giving shorter RCT and increased curd firmness, the effects on curd firmness being more pronounced. The effect of κ-casein variant on RCT could be eliminated by reducing pH and adding CaCl2 to the milk samples. The effect on curd firmness was not altered by this treatment, however. The results show that the renneting properties of individual milks are to a considerable degree genetically controlled.

N-acetyl-β-D-glucosaminidase (NAGase) levels and somatic cell counts (SCC) were determined monthly for 6 months in the bulk milk of 181 suppliers (1063 samples). A highly significant correlation (r = 0·74; P < 0·001) was found between supplier's bulk herd milk geometric mean NAGase activity and SCC. Monthly trends which grouped suppliers into various categories defined by different NAGase and SCC thresholds showed that a similar overall pattern was obtained with both SCC and NAGase. However, further analysis indicated that 18% of the bulk milk which had SCC < 500 × 103/ml had NAGase levels > 25 units. Also, 34% of samples with SCC > 500 × 103/ml had NAGase levels < 25 units. Overall, 24% of all samples did not have corresponding NAGase and SCC results. When the bulk milk of 2 commercial dairy herds was tested monthly over 12–18 months whilst the infection status of all quarters in the herds was regularly monitored, those herds with low incidence of mastitis (5% quarters infected) had significantly lower bulk milk SCC and NAGase levels than those with a high incidence of mastitis (22% of quarters infected). This suggests that NAGase measurement on bulk herd milk would be a simple means of monitoring infection status of dairy herds and of rapidly classifying a supplier's milk as being of low, medium or high SCC status. The possible combined use of SCC and NAGase levels in bulk milk monitoring schemes is discussed.

The composition and weight of deposit formed in all sections of an ultra-high-temperature milk sterilization plant were determined. Deposits formed in the preheating, heating and cooling sections during sterilization of pasteurized whole milk were analysed for dry matter, protein, fat and mineral contents. The weight and composition of components of the deposit varied in different sections of the plant and with the heating temperature. Two categories of deposit could be distinguished: one in the preheating section consisting of (w/w) protein 50%, minerals 40% and fat 1% and another in the heating section consisting of minerals 75%, proteins 15% and fat 3%. Concerning the rate of formation of these deposits, a comparison of the results with those obtained for pasteurization shows that fouling was more rapid during pasteurization of raw milk than during sterilization of previously pasteurized milk. Hypotheses concerning mechanisms of formation of protein deposition are discussed.

An attempt was made to partition changes in milk yield during lactation in the rat between changes in the size of the secretory cell population and changes in cellular activity. Groups of rats were studied on d 2, 7, 14, 21 and 28 of lactation. Milk yield increased between d 2 and 7, less markedly between d 7 and 14 and decreased between d 14 and 21. Milk secretion had apparently ceased on d 28. Total mammary DNA (DNAt) and RNA (RNAt) increased from d 2 to 14. It is concluded that of the increase in milk yield between d 2 and 14, an increase in cell population accounted for 75% and an increase in cellular activity for 25%. A decrease in activity appeared to be entirely responsible for the decrease in yield between d 14 and 21.