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.

Experiments were undertaken to validate a method (using adrenaline injection) for determination of the size of cisternal and alveolar compartments in the udder, to use this method to determine the pattern of milk accumulation in the udder over time and to determine the relationship between the size of the alveolar and cisternal compartments and tolerance of once daily milking. Cows received intrajugular injections of adrenaline (3 mg) immediately before milking, to block milk ejection and allow harvesting of the cisternal milk fraction. This was followed by removal of the alveolar fraction 30 min later after intrajugular oxytocin (5 i.u.) injection. Results obtained were similar to those obtained by catheter drainage. The alveolar compartment was 90% full at 16 h post milking while the cisternal compartment filled more slowly and was only 70% full at 24 h post milking. At full capacity (measured at 40 h), the volumes of milk contained in the cisternal and alveolar compartments were similar. In a further experiment involving identical twin cows, it was shown that the greater the degree of filling of the cisternal compartment at 24 h, the lower was the production loss on once daily milking. This suggests that the freedom of the alveoli to drain was an important factor in the production loss on once daily milking. Although there were significant correlations within twin sets for milk yield and the size of udder compartments, the relationship within twin sets for yield loss on once daily milking was not statistically significant.

Records of claw trimmings were analysed in seven organic and six conventional Danish herds (a total of 974 cows). The housing systems represented were tie stall systems, loose housing system with slatted floor (one organic herd), and deep litter systems (deep straw bedding). Occurrence of sole disorders was analysed separately for cows in first lactation and for cows in later lactations. Three different responses (acute haemorrhage, sole ulcer in one leg and sole ulcer in two or more legs) were analysed using three binomial logistic regression analyses for each group. Herd analysed as a fixed effect was a strong risk factor for all kinds of sole ulcer. Lactation stage was a risk factor for acute haemorrhage in both groups of cows, and for sole ulcer in first parity cows. In general, there was a strong positive association between the period 61–120 d post partum and the presence of sole disorders. Breed was associated with acute haemorrhage in cows in second and later parities, and sole ulcer in one leg only in first parity cows in an interaction with lactation stage in both conditions. Danish Friesian cows were strongly associated with sole disorder, although the combination of lactation stage from 61 to 120 d post partum in cows of other dual purpose breeds was positively associated with the presence of sole ulcer in one leg only in first parity cows. The time of year for claw trimming was a risk factor for acute haemorrhage in first parity cows, with the period from December to January most strongly associated with acute haemorrhage. Previous disease treatment was a risk factor for sole ulcer in two or more legs in second and later parities. Udder related disorders and disorders other than reproductive problems were positively associated with the occurrence of sole ulcer. Body weight at calving was associated with acute haemorrhage in cows in second and subsequent parities. Body weight lower than the mean herd level by >50 kg was negatively associated with acute haemorrhage.

We found previously that the current recommendations for Na+, K+, and Cl− contents in the diet do not meet the needs of lactating cows. The responses of cows receiving a ration with increased amounts of Na+, K+, and Cl− (E cows) were compared with those of cows consuming the same ration with a fixed concentration of these ions (C cows) between weeks 2 and 8 post partum (PP). Milk, protein, fat and lactose yields, and dry matter intake between weeks 2 and 4 PP were higher in E than in C cows. These differences did not occur between weeks 4 and 8 PP, mainly because of a higher incidence of PP complications in E cows. A greater increase in plasma insulin-like growth factor-1 concentration in E than in C animals during weeks 2 and 3 PP was consistent with the milk responses. A reduction in aldosterone concentration in E cows in weeks 2 and 3 PP was a consequence of their Na+ requirements being satisfied as a result of their enhanced Na+ intake. A subsequent elevation in aldosterone concentration in E animals was probably related to a moderate excess in K+ intake. This increase in aldosterone explains the urinary potassium loss that was detected at week 6 PP. The absence of differences between E and C cows in plasma renin activity was consistent with an absence of differences in urine volume and with the apparent utilization of the enhanced ion intake for body functions.

Forty-two multiparous dairy cows of three different breeds (Holstein, Montbéliarde and Tarentaise) were fed on the same type of forage (natural grassland) preserved in the form of either hay (H) or silage (GS), according to a changeover design (two 4 week periods). The proportion of concentrate in the diet and the energy and nitrogen contents were similar in both treatments. The milk produced by these cows was used for the manufacture of Saint-Nectaire type cheeses, under controlled and identical cheesemaking technological conditions. More cheese was produced with the H treatment milk. The cheeses made with the GS treatment milk were more yellow and tended to be more bitter. The other chemical and sensory characteristics did not differ much between the two treatments. Of the 51 volatile compounds identified, four were in significantly higher proportion in the GS than in the H cheeses. Cheeses produced from Tarentaise cows' milk were more yellow and their pH was higher than those made with the milk of Holstein or Montbéliarde cows. The cheeses from Montbéliarde and Tarentaise cows' milk were firmer, more melting and tastier than those made with the milk of Holstein cows. Although some trends were apparent, there were no significant differences in cheese volatile compounds for different breeds.

In order to describe the kinetics of rennet coagulation, measurements of turbidity as a function of wavelength were used to determine the weight-average degree of polymerization, x¯w, during renneting of milk at three different concentrations of enzyme and three concentrations of casein, including the normal casein concentration of milk. The change of x¯w as a function of time was described using Von Smoluchowski's equation, testing a number of expressions for the aggregation rate constant, kij. The best description was achieved when kij was taken as a function of an energy barrier against aggregation that was diminished by the proteolysis of κ-casein. The initial value of the energy barrier partly depended on the casein concentration, and had a value >25 kBT at normal casein concentration, where kB is Boltzmann's constant and T the absolute temperature. When the proteolysis of κ-casein was complete, the energy barrier was reduced to 11 kBT and was independent of casein concentration.

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.

Cows with subclinical intramammary infections were identified by milk bacteriology. The mastitis pathogens included Staphylococcus aureus (n=9), Streptococcus uberis (n=10) and coagulase-negative staphylococci (n=10). Samples of first fore milk, main flow milk and strippings milk fractions were collected from each quarter and laboratory measurements were made of electrical conductivity, milk fat concentration and somatic cell count. Conductivity measurements were corrected for milk fat concentration and within-cow inter-quarter conductivity ratios calculated. Repeatability estimates of all measurements between days were calculated. In the case of infected quarters, all conductivity values decreased markedly (P<0·05) from first fore milk to main flow milk fractions. Conductivity differences between quarters of infected cows were substantially lower during the main milk flow phase. For quarters infected with Staph. aureus an increase in conductivity was observed (P<0·05) from main flow to strippings fractions. For uninfected quarters, conductivity declined as milk fat concentration increased with successive milk fractions. Variation, both within and between milk fractions, was greater for somatic cell count than for conductivity. Differences in conductivity between milk fractions from individual infected quarters were not accounted for by changes in fat concentration and may result from the mixing of milk from infected and uninfected regions of the gland. Localized infection may produce a decrease in conductivity between fore milk and mid-flow fractions while differential drainage from an infection site in the secretory tissue may additionally produce an increase in conductivity from mid-flow to strippings fractions. Such changes may thus provide information on the location and magnitude of an infection. The results clearly demonstrate the importance of the milk fraction when using conductivity as a diagnostic of intramammary infection, the highest diagnostic sensitivity being achieved by using first fore milk samples.

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 interactions of denatured whey proteins with casein micelles on the rheological properties of acid milk gels was investigated. Gels were made by acidification of skim milk with glucono-δ-lactone at 30°C using reconstituted skim milk powders (SMP; both low- and ultra-low-heat) and fresh skim milk (FSM). The final pH of the gels was ∼4·6. Milks containing associated or ‘bound’ denatured whey proteins (BDWP) with casein micelles were made by resuspending the ultracentrifugal pellet of heated milk in ultrafiltration permeate. Milks containing ‘soluble’ denatured whey protein (SDWP) aggregates were formed by heat treatment of an ultracentrifugal supernatant which was then resuspended with the pellet. Acid gels made from unheated milks had low storage moduli, G′, of <20 Pa. Heating milks at 80°C for 30 min resulted in acid gels with G′ in the range 390–430 Pa. The loss tangent (tan δ) of gels made from heated milk increased after gelation to attain a maximum at pH ∼5·1, but no maximum was observed in gels made from unheated milk. Acid gels made from milks containing BDWP that were made from low-heat SMP, ultra-low-heat SMP and FSM had G′ of about 250, 270 and 310 Pa respectively. Acid gels made from milks containing SDWP that were made from ultra-low-heat SMP or FSM had G′ values in the range 17–30 Pa, but gels made from low-heat SMP had G′ of ∼140 Pa. It was concluded that BDWP were important for the increased G′ of acid gels made from heated milk. Addition of N-ethylmaleimide (NEM) to low-heat reconstituted milk, to block the —SH groups, resulted in a reduction of the G′ of gels formed from heated milk but did not reduce G′ to the value of unheated milk. Addition of 20 mm-NEM to FSM, prior to heat treatment, resulted in gels with a lower G′ value than gels made from reconstituted low-heat SMP. It was suggested that small amounts of denatured whey proteins associated with casein micelles during low-heat SMP manufacture were probably responsible for the higher G′ of gels made from milk containing SDWP and from milk heated in the presence of 20 mm-NEM, compared with gels made from FSM.

The transport of L-glutamine by the lactating rat mammary gland has been investigated using rat mammary tissue explants and the in situ perfused rat mammary gland. L-glutamine uptake by both explants and the perfused mammary gland was via both Na+-dependent and Na+-independent pathways. It appeared that these pathways are situated on the blood-facing aspect of the mammary gland. L-glutamine uptake by both mammary preparations was markedly inhibited by 2-aminobicyclo[2,2,1]heptane-2-carboxylic acid in the absence of external Na+. This is consistent with L-glutamine uptake via system L. The Na+-dependent component(s) of L-glutamine uptake remains to be precisely identified. However, system A can be ruled out on the basis that L-glutamine was not inhibited by (methylamino)isobutyric acid. Mammary tissue concentrates L-glutamine with respect to both milk and plasma: we suggest that the Na+-dependent component(s) of L-glutamine uptake is responsible for generating the intracellular to extracellular concentration gradient.

Milk was collected from three spring-calving herds, on different daily herbage allowances (DHA) of perennial rye-grass (16, 20 or 24 kg dry matter (DM)/cow for a 17 week period. On five occasions, at weekly intervals in the middle of the period, the three different milks were converted into low-moisture part-skim Mozzarella cheese. Increasing the DHA resulted in significant increases in the concentrations of protein in the cheesemilk (P<0·05) and cheese whey (P<0·02). The moisture-adjusted cheese yield increased significantly (P<0·01) on raising the DHA from 16 to 24 kg grass DM/cow. DHA had no significant effects on any of the gross compositional values of the cheese (although moisture and fat-in-DM levels tended to decrease and increase respectively with increasing DHA). The hardness of the uncooked cheese and functionality of cooked cheese (i.e. melt time, flowability, stretch and viscosity) were not significantly influenced by DHA over the 115 d ripening period at 4°C.

Concurrent falls in milk production and electrical resistance of composite milk were examined in Israeli Holstein cows. The cows were milked three times a day by a system that recorded yield and the lowest electrical resistance in the composite milk from the four glands. The study included two groups: cows that experienced on day 0 a decline in resistance and milk production [ges ]20% from the mean of the previous 9 d (62 cows, case group) and cows that experienced no such episodes over 9 d before and after a fixed day (118 cows, control group). Bacteriological status and somatic cell count (SCC) or California mastitis test scores were assessed on the fixed day in the control group, and on days 0, 1 and 2 in the case group. California mastitis test scores greater than 2 and SCC thresholds of 5×105 cells/ml were used to create two classes of leucocytosis. There was no statistically significant difference between the two groups in frequency distributions of pathogens and their types: in 30% of cows infection was not detected, 33% were infected by major pathogens (95% of which were Staphylococcus aureus), and 53·5% by minor pathogens (80% Micrococcus spp.). Cows in the case group had lower milk production during the 8 d following day 0. Mean electrical resistance was lower in infected cows and particularly in cows infected by Staph. aureus. High leucocytosis was associated with reduced electrical resistance in both groups, and was found in 93% of cows in the case group v. 25% in the control group. The results suggest that falls in electrical resistance of milk and in milk production were not linked to a specific pathogen, and were followed by 3–8 d of reduced milk production and electrical resistance. The study suggests that there are episodic aggravations in mammary health that do not evolve into clinical mastitis but may induce significant losses in milk yield and quality.

αs1-, αs2-, β- and κ-caseins from Somali camels (Camelus dromedarius) were purified by acid precipitation at pH 4·4, crudely separated into an α-CN and a β-CN fraction and further purified by reversed-phase HPLC. Fragments of tryptic digests were sequenced. Amino acid patterns obtained were used to screen a cDNA library constructed from mRNA from lactating udder tissue. Full length clones corresponding to the four caseins were sequenced. The numbers of residues in the sequences deduced were αs1-CN 207, αs2-CN 178, β-CN 217, κ-CN 162. Percentage similarity to bovine proteins was αs1-CN A 39, αs2-CN 56, β-CN 64, κ-CN 56. Acid-precipitated casein of pooled milk was separated by reversed-phase HPLC and monitored at 220 nm, and its composition, estimated from peak integration, was (g/kg total casein) αs1-CN 220, αs2-CN 95, β-CN 650, κ-CN 35. Degrees of phosphorylation and glycosylation were determined by laser ionization mass spectrometry and sequence pattern analysis. Molecular masses determined were (kDa) αs1-CN A, 24·755 and 24·668; αs1-CN B, 25·293; αs2-CN 21·993; β-CN, 24·900; κ-CN 22·294–22·987. The pH values of the most probable isoelectric points were: αs1-CN A 6P 4·41, αs1-CN B 6P 4·40, αs2-CN 9P 4·58, β-CN 4P 4·66, κ-CN 1P, with ten sialic acid residues bound, 4·10.

Digestion of bovine αs1-casein with bovine trypsin produced peptide(s) with an inhibitory effect on concanavalin A-induced proliferation of mouse spleen cells. One of these peptides was isolated from the αs1-casein digest following ultrafiltration, hydroxyapatite chromatography and reversed-phase HPLC, and amino acid composition and sequence analyses showed it to be sequence 59–79 from the phosphoserine-rich region of αs1-casein. The isolated peptide significantly inhibited the concanavalin A-induced proliferation of mouse spleen cells and rabbit Peyer's patch cells, whereas it enhanced the lipopolysaccharide- and phytohaemagglutinin-induced proliferation of both cells. The peptide displayed mitogenic activity in the cell cultures without the commercial mitogen, and significantly enhanced immunoglobulin production. Moreover, residues 1–25 from the phosphoserine-rich region of bovine β-casein had a similar effect on the proliferation of mouse spleen cells and rabbit Peyer's patch cells stimulated or not stimulated by the commercial mitogen. These results indicate that caseinophosphopeptides may act as a humoral immunostimulator in cell cultures.

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.

The aim of this study was to determine whether carbonic anhydrase (CA) activity in goat mammary capillaries is regulated mainly by local or systemic mechanisms. One gland was dried before the contralateral gland, and after parturition only one gland was milked. Biopsies were taken from the mammary glands of three goats at 14 d intervals during involution and the start of the following lactation. A histochemical method was used to visualize sites of CA activity. To follow the involution process, milk (liquid) samples were taken from both teats each week and analysed for pH and composition. The time course of CA activity disappearance and reappearance in the capillaries was related to changes in milk composition and alveolar area. A dense network of capillaries showing membrane-bound staining for CA was found surrounding the alveoli in the lactating gland. CA activity gradually decreased in the drying gland, although the other gland was being milked. After 8 weeks involution the dried gland had a significantly lower number of stained capillaries than the milked gland. Almost no stained capillaries were found during late pregnancy, when both glands were dried and the tissue growth maximal. During lactation milk pH was 6·6±0·3 and this increased to 7·0±0·1 in the course of involution. In the last trimester of pregnancy the pH returned to its lower value, while the mammary gland was devoid of stained capillaries. Therefore, the capillary CA could not have been directly involved in the pH regulation of milk. The CA activity reappeared in the capillaries directly after delivery, but only in the milked gland. Clearly the regulation of CA activity is influenced more by local than by systemic factors and is associated with the metabolic activity of milk secretion.

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.

Bovine αs1-casein F (αs1-CN F) was found in a genetic resource of Deutsches Schwarzbuntes Niederungsrind cows at a frequency of 0·009. Biochemical characterization of this new variant was obtained by automated sequencing of reversed-phase HPLC-separated tryptic peptides of αs1-CN F and αs1-CN B. αs1-CN F was found to be a subtype of αs1-CN B with a single amino acid substitution (SerP/Leu) in position 66. DNA sequencing revealed a C/T transition in position 8418 of the gene. Sequence-specific primers were designed to perform an allele-specific polymerase chain reaction for detection of αs1CnF. Typing of artificial insemination sperm samples included in the genetic resource sperm pool identified one sire heterozygous for αs1CnF.

Homocitrulline arises from the reaction between cyanate and the ε-amino group of lysine residues; in milk, cyanate derives from heat-induced urea breakdown. Since homocitrulline levels were unknown in heated milk, its formation was studied in the temperature range 100–150°C. Firstly, an analysis method based on ion-exchange chromatography using an amino acid analyser was developed. Homocitrulline, liberated from milk protein by enzymic hydrolysis, was eluted well separated from other amino acids and could be readily distinguished using this technique. Secondly, homocitrulline levels were determined for various time–temperature combinations in samples of milk, milk to which 10 mm-urea had been added, and milk that was made urea-free. No homocitrulline was formed in urea-free milk, while homocitrulline formation was stimulated by urea addition. Upon prolonged heating, we found extensive subsequent breakdown of homocitrulline. The kinetics of homocitrulline formation was quite complicated, but an approximation was possible as the initial formation reaction could be modelled using zero order reaction kinetics. The apparent activation energy for homocitrulline formation was estimated at ∼90 kJ/mol. In general, the levels of homocitrulline to be expected in heated milk appeared to be quite low: ∼0·3 mm in in-bottle sterilized milk, and <0·01 mm in UHT sterilized milk.