The effect of storing Cheddar cheeses at elevated temperatures on proteolysis and flavour development was examined. Cheeses were stored under the following ripening conditions: (1) 8 °C for 32 weeks (control); (2) 15 °C for 8 weeks then 8 °C for 24 weeks; (3) 17·5 °C for 8 weeks then 8 °C for 24 weeks; (4) 20 °C for 8 weeks then 8 °C for 24 weeks; (5) 15 °C for 32 weeks; (6) 17·5 °C for 32 weeks; and (7) 20 °C for 32 weeks. The rates of build up of products of proteolysis and flavour increased in the order of the above treatments and substantial acceleration of ripening was achieved. However, after 32 weeks at 17·5 °C and 16 weeks at 20 °C, cheeses received significantly lower preference scores than the control cheeses (P < 0·05) mainly owing to the development of off flavours. The maximum temperature at which cheeses could be stored for 32 weeks without reduction in quality was 15 °C. Cheeses stored at this temperature for the entire period had levels of products of proteolysis at 12 weeks of age which were similar to those of controls at 32 weeks, and estimations of age at 20 weeks which were similar to those of controls at 32 weeks.

Differences in flavour scores and in the concentrations of free fatty acids, methyl ketones and H2S were measured in Cheddar cheeses containing various groups of non-starter bacteria or starter streptococci alone, made under controlled bacteriological conditions by the aseptic vat technique. The non-starter bacteria were made up of lactobacilli, leuconostocs, pediococci, micrococci and Gram-negative rods isolated in commercial creameries from raw milk or fresh cheese curd. These were added to the experimental cheese as single groups or as complete floras (reference floras). Several bacterial groups influenced the measured concentrations of the flavour compounds, but flavour differences were not correlated with these chemical differences. Only cheese containing a curd-derived whole reference flora or cheese made in open vats in the N.I.R.D. Experimental Dairy had significantly better flavour than starter-only cheese, but this improvement was not attributable to any particular group of bacteria.

Rheology is denned as the science of the deformation of matter. It includes the study of all deformations, recoverable (elastic) and non-recoverable (plastic). In this paper cheese is primarily considered although the fundamental principles apply to all materials.

The elastic and plastic properties of cheese exercise a great influence in the ripening processes and the incidence of faults. Although much progress has been made in the bacteriological and chemical investigations of cheese ripening, little fundamental work on the nature of and the factors controlling the physical properties of curd and cheese has been carried out.

The objective was to compare the efficacy of two experimental Staphylococcus aureus mastitis bacterins and a currently marketed five-isolate-based Staph. aureus bacterin (Lysigin™, Boehringer Ingelheim Vetmedica, Inc.) with unvaccinated controls. Forty-seven Holstein-Friesian heifers were randomly assigned to one of four groups such that Group 1 (n=11) received a three-isolate experimental bacterin, Group 2 (n=11) received a five-isolate experimental bacterin, Group 3 (n=14) received Lysigin, and Group 4 (n=11) served as unvaccinated controls. Vaccinations were administered twice 28 d apart in late gestation. All groups were challenged with a heterologous strain of Staph. aureus (ATCC 29740) on days 6, 7, and 8 of lactation. Mastitis score, somatic cell count (SCC), milk culture yield, and total daily milk yield data were collected before and after challenge. All 47 cattle developed a Staph. aureus IMI post-challenge with three animals in Group 1 and one animal in Group 3 clearing their Staph. aureus IMI by the end of the study. However, there was no evidence of a difference between vaccinates and control with regard to Staph. aureus clearance rates post-challenge (P[ges ]0·214). Cattle vaccinated with Lysigin had a lower mean duration of clinical mastitis and lower total mastitis score post-challenge than controls (P=0·045 and P=0·046, respectively). Overall, there was no evidence that any of the vaccinated groups had a lower mean SCC than control (P[ges ]0·148) for the tested study days. Likewise there was no evidence that vaccinates had greater milk yield than controls post-challenge (P=0·617). Hence, there was no evidence that the vaccines reliably prevented Staph. aureus IMI, but Lysigin showed benefit in reducing the clinical severity and duration of clinical disease post-challenge. Neither of the experimental bacterins appeared to perform better than Lysigin.

A total of 1304 goat udder halves were sampled monthly during an entire lactation (6262 samples) with the aim of identifying factors affecting milk somatic cell count (SCC). Bacteriological analyses for identification of mastitis pathogens were carried out on all samples and SCC was also determined. All animals were examined for infection by caprine arthritis-encephalitis virus (CAEV) using a commercial ELISA test kit. Results obtained were arranged in two databases (whole-lactation average half-udder database and monthly half-udder database) and two mixed models were applied. Random effects of half udder nested into flock and fixed effects of flock, intramammary infection (IMI) status, number of kids born, length of lactation and interaction of parity with IMI status were significant for the first database. CAEV infection and its interaction with IMI status was not significant. Milk SCC was significantly increased for infected udder halves and milk from udder halves infected with minor pathogens had lower SCC than udder halves infected with major pathogens. For healthy udder halves, SCC was higher in older animals but this effect was not evident in halves with IMI. Multiple birth and short-duration lactation were factors associated with elevated milk SCC. The second mixed model considered repeated measures in time for consecutive samplings throughout lactation (stage of lactation) which was also a significant factor with increasing stage of lactation. The influence of all these factors should be taken into account in the establishment of more reliable diagnostic SCC thresholds for IMI.

The possible hypocholesterolaemic properties of milk and fermented milk products have been investigated in groups of albino rats given a basal diet, basal diet plus cholesterol, and basal diet plus cholesterol together with whole milk or standard or bifidus yogurt. The yogurts were fortified with skim milk powder, condensed whey or lactose-hydrolysed condensed whey. After 30 d, triacylglycerols, total cholesterol, HDL-cholesterol and LDL-cholesterol were measured in serum. Whole milk and ordinary yogurt had no hypocholesterolaemic effect, but standard yogurt containing lactose-hydrolysed condensed whey and all bifidus yogurts lowered serum cholesterol. In general, yogurts changed HDL-cholesterol little, but tended to raise triacylglycerols. There was marked lowering of LDL-cholesterol in rats given either type of yogurt fortified with whey proteins. This study has demonstrated in a rat model that bifidus yogurts and yogurts fortified with whey proteins can reduce total and LDL-cholesterol, and suggests that if they have the same effect in human subjects they have potential value in cholesterol-lowering diets.

Aseptic udder quarter secretion samples (n = 1112) from 200 mastitic and 65 non-mastitic control heifers from 160 different farms were collected for bacteriological examination and in vitro antimicrobial susceptibility testing. Coagulase-negative staphylococci (CNS) were the most frequently isolated bacteria (57·8%). The percentage of Staphylococcus aureus was 20·1, of streptococci 11·3 and of other pathogens 10·8. Staph. simulans, Staph. hyicus, Staph. xylosus and Staph. chromogenes were the most common CNS. Staph. simulans and Staph. hyicus were most frequently found in clinical samples after parturition, whereas other CNS were equally or more often found in control samples. Most CNS infections detected in the prepartum period were eliminated spontaneously or with antibiotic treatment during early lactation, but the infected quarters were more susceptible to new infection by other pathogens. Milk N-acetyl-β-glucosaminidase activities were measured to monitor changes in inflammation. Among the CNS infections, N-acetyl-β-glucosaminidase activities were highest for Staph. hyicus and lowest for Staph. xylosus in the lactating mammary gland of heifers. CNS were resistant to trimethoprim–sulphonamide, ampicillin and erythromycin. Staph. aureus was more resistant than CNS to β-lactamase-sensitive penicillins.

Milk which had been heated to 85°C for 40 min was examined for changes to milk salts taking place upon cooling to 4, 20, 40 and 60°C. The time-concentration curves showed a two-step reaction for Ca, Pi and pH. The effect of the heat treatment in milk heated to 85°C for 40 min was to produce greater changes in the second step of the reaction. Because of the variability of the results for other species (Mg and citrate), the reversibility was calculated only for Ca and P. It was 90–95% and 93–99% respectively depending on the cooling temperature. The reversibility of the heat-induced changes and especially of the possible dissolution equilibrium between Ca phosphate and citrate precipitates on cooling are discussed.

Cation-exchange chromatography on a Mono S column (Pharmacia) was used to separate macropeptide from whey proteins. Macropeptide was eluted by 0·1 M-NaCl in a 20 mM-KCl–HCl buffer, pH 2. This technique was suitable for quantitative determination of macropeptide in rennet whey and also for following the action of chymosin on κ-casein in skim milk. Precipitation at pH 4·6 was used to remove residual caseins and to keep macropeptide in solution. In comparison with other methods for determining macropeptide, the present one eliminates the need for pretreatment of samples with trichloroacetic acid (TCA) and allows the recovery of all the macropeptide. Quantitative determination of macropeptide in the 8% TCA-soluble fraction by cation-exchange chromatography showed that only 50–75% of the macropeptide was recovered. This chromatographic technique could also be applied for isolating and producing whole macropeptide on a preparative scale.

The incorporation of serum proteins into the synthetic fat globule membrane in milks that had been heated and subsequently homogenized has been quantitatively determined using electrophoresis and densitometry. The results indicated that the amounts of serum proteins present in the fat globule membrane were smaller in milk that was heated before homogenization than in milk that had been homogenized and then heated. The thickness of the adsorbed layer of protein on the surfaces of the fat particles, measured using photon correlation spectroscopy in dissociating and non-dissociating buffer solutions, was also lower in the former case (∼ 20 nm) compared with the latter (∼ 34 nm). The kinetics of adsorption of both β–lactoglobulin and α-lactalbumin in heated homogenized milk on the vacant ‘sites’ on the synthetic fat globule membrane followed similar trends at 75 and 90 °C.

The effects of supplementing a basal diet, in which the low level of fatty acids limited milk production, with soya oil, a palm oil/palmitic acid mixture and tallow, on the yield of milk and of its constituents, and on the composition of the milk, are reported. The yields of milk and milk fat were greatly increased by all the oil-supplemented diets; the mean daily yield of solids-not-fat (SNF) was also increased, but supplementation with soya oil caused the yield of crude protein (CP) to decrease, whereas the other fat-rich concentrate mixtures gave the same mean yield of protein as did the low-fat, control diet. All 3 oil-supplemented diets lowered the proportion of CP in the milk, but the SNF content was unchanged by any treatment. Dietary soya oil tended to lower the proportion of fat in the milk, whilst the palm oil/palmitic acid mixture raised it, with the tallow exerting no effect. The results are discussed in relation to previous work in which these dietary oils have been used, but in which the intake of fatty acids from the control diets did not limit milk production to the extent reported here.

A sandwich ELISA (enzyme-linked immunosorbent assay) of the two-site type has been successfully developed for the detection of cows' milk in goats' or ewes' milk. The assay uses two monoclonal antibodies (MAb) raised in mice against cows' β-lactoglobulin (β-lg). These MAb recognize different epitopes of the β-lg, which are sufficiently distinct to allow simultaneous binding of the corresponding antibodies. One of the MAb recognizes a species-specific epitope of the bovine β-lg and was adsorbed to a plastic microtitration plate (capture antibody). The second MAb was labelled with peroxidase and used to detect the captured cows' β-lg. Factors affecting assay performance were investigated. The optimized assay is highly specific, reproducible (intra- and inter-assay CV were 8 and 13% respectively) and sensitive: as little as 5 ng β-lg/ml or 1 part cows' milk per 100000 parts goats' or ewes' milk can be detected. The technique is robust, cheap, rapid, reliable and suitable for high sample throughput, semi-automation and screening surveys. The MAb used guarantee the high specificity of the assay and indefinite reagent supply of constant quality once approved by collaborative national or international trials.

The effect of bacterial infections, produced by infusions of Streptococcus dysgalactiae and of Staphylococcus pyogenes into one quarter of the udder, on milk yield and composition in heifers has been investigated. After a quarter became infected there was invariably a decrease in the yield of milk and in the concentrations of lactose and potassium, while there was an increase in those of sodium, chloride and non-casein proteins. These effects persisted into the 2nd lactation if the infection was not eliminated. If the infection was eliminated during the 1st lactation or during the dry period before 2nd calving there was a complete recovery in the composition of the milk in the 2nd lactation, but the recovery in milk yield was not complete.

Peptide and amino acid transport in Streptococcus lactis was both energy-and temperature-dependent. Separate systems were present for the transport of amino acids, dipeptides and oligopeptides as well as for individual amino acids. Both tri- and tetra-peptides were readily transported while an apparent size restriction at the point of uptake resulted in a relatively low rate of uptake of the pentapeptide. The effects of pH, temperature and salt concentration on the uptake of amino acids and peptides were studied. Exchange reactions played a significant role in the uptake of both peptides and amino acids.

The aim was to determine whether administration of melatonin would alter the yield and composition of milk from grazing dairy cows in summer. Twelve sets of spring-calving identical twin Friesian cows were used in the experiment. In late-November (late spring), one twin from each set was given slow-release melatonin implants behind the ears (108 mg melatonin/cow). Two further implantations occurred at 4-weekly intervals to maintain increased circulating concentrations of melatonin for 12 weeks. The other twin served as a control. Milk yield and composition were measured twice prior to treatment and then four times over the following 12 weeks. Concentrations of melatonin, prolactin and insulin-like growth factor 1 (IGF-1) were measured in blood plasma twice before treatment and then either seven (melatonin and prolactin) or three (IGF-1) further times during the experiment. Management procedures for all cows were similar and cows grazed a daily pasture allowance of approximately 30 kg DM/cow as their sole feed source. In melatonin-treated cows there was a decrease in mean concentrations of prolactin in plasma, but concentrations of IGF-1 did not change. Melatonin reduced milk yield by 6 weeks after treatment and by the end of the 12-week experimental period milk yield in melatonin-treated cows had fallen by 23%. Melatonin also reduced concentrations of lactose in milk, but increased concentrations of fat, protein and casein, changes that were broadly similar to those that occur in late lactation in seasonally calving dairy cows. Thus, the results suggest that some of the variation in the volume and quality of milk throughout the season in New Zealand dairy systems may be due to changes in photoperiod mediated by increased concentrations of plasma melatonin in association with decreased concentrations of plasma prolactin.

The effect of physicochemical characteristics (pH, temperature, composition) of the aqueous phase on the mineral balance in milk and milk retentate has been studied. The ratio of colloidal Ca to total protein decreased with pH, but at any given pH the higher the protein concentration, the higher was the ratio of colloidal Ca to total protein. The solubilization of Ca during cooling and the decrease in soluble Ca during heating were approximately the same in retentates and in milk. Among the components of the aqueous phase, soluble Ca and citrate ions were related to the amount of colloidal Ca.

The distribution and activity of alkaline phosphatase (E.C. 3.1.3.1), acid phosphatase (E.C. 3.1.3.2), catalase (E.C. 1.11.1.6), xanthine oxidase (E.C. 1.2.3.2), aldolase (E.C. 4.1.2.7 and 4.1.2.13), ribonuclease (E.C. 2.7.7.16) and carbonic anhydrase (E.C. 4.2.1.1) were studied in the major components of bovine milk. Fractionation was accomplished by centrifugation of milk, skim-milk and buttermilk, and ammonium sulphate precipitation of skim-milk serum. The range of activities found for the enzymes studied are tabulated together with the activities of some of the enzymes in mastitic milk, and the significance of the results obtained is discussed. No carbonic anhydrase activity was detected in any of the samples tested. The other enzymes studied were found to have a greater proportion of their total activity located in the skim-milk fraction. However, all of these enzymes except ribonuclease had a higher specific activity in the fat fraction.

1. A statistical analysis has been made of lactation records from 500 dam-daughter pairs of the Ayrshire breed. Measurements were taken of milk yield, fat, S.N.F., crude protein, casein and lactose.

2. A correction was made for the age of the cow. This contributed a substantial fraction of the total variation for milk yield, S.N.F. and lactose contents. Month of calving had a trivial effect on the lactation average for the characters when compared with the individual variation between cows.

3. After correction for age, the variation between lactations was highest for fat content (standard deviation 0·35%) and slightly less for S.N.F. (0·26%).

4. The heritability was estimated by the dam-daughter correlation within herds. Values in the neighbourhood of 0·5 were obtained for S.N.F., crude protein, casein and ‘ash’ (determined by difference), about 0·35 for fat and lactose, 0·25 for milk yield and 0·13 for non-casein nitrogen.

5. The phenotypic correlations between yield and lactose were positive for both dam and daughter groups. The correlations between yield and the other constituents were more negative in the daughter group than in the dam group. The correlation between yield and S.N.F. was, in fact, positive in the latter. The correlation between fat and S.N.F. averaged 0·36, being derived mainly from the correlation between fat and casein (0·43), though that with lactose was also positive (0·13).

6. The variation in S.N.F. was mostly determined by that in casein (r=0·85). There was a positive correlation between lactose and casein (0·24), and a negative one between lactose and non-casein nitrogen (–0·41). Lactose was negatively correlated with ‘ash’ (–0·31). ‘Ash’ was positively correlated with non-casein nitrogen (0·23)

7. There were positive and significant genetic correlations between casein and both fat and lactose, and that between the last two was almost significant.

8. No significant genetic correlations were found between yield and the chemical constituents, although figures of practical importance may exist which did not reach significance in an experiment of this size.

9. The practical significance of these results is discussed. It is concluded that selection for yield alone is not likely to cause any great decline in S.N.F. content of the milk, and that this could probably be held in check by paying some attention to the fat content of the milk.

Lactoperoxidase activity and thiocyanate content were monitored in the milks of seven individual cows and six goats during lactation. Lactoperoxidase activity exhibited a cyclic pattern with alternating peaks and troughs throughout lactation. Extremely large variations were observed between and within animals. For example, lactoperoxidase activity ranged from 0·05–5·60 U/ml for one cow on different sampling days. Variations between cows were also large, ranging from 0·98–5·1 U/ml on a specific sampling day. Mean lactoperoxidase activity in cows ranged from 1·5–2·7 U/ml with an overall mean of 2·3±1·0 U/ml while the thiocyanate concentration ranged from 6·0–10·2 μg/ml with an overall average of 8·5±5·1 μg/ml. Lactoperoxidase activity means for goats ranged from 0·04–0·16 U/ml with a grand mean of 0·1±0·06 U/ml while the thiocyanate content means ranged from 6·6–8·2 μg/ml with a grand mean of 7·0±2·59 μg/ml. There was no correlation between lactoperoxidase activity and thiocyanate content in either cows' (R2 = 0·011) or goats' (R2 = 0·015) milk. These experiments have revealed that lactoperoxidase activity is affected by many factors including the individual animals, species, feed and stage of lactation. Therefore the exogenous supply of thiocyanate and hydrogen peroxide needed to activate the lactoperoxidase system for raw milk preservation will vary in quantity depending on these factors.

Intracellular peptidases of Lactobacillus helveticus may play a major role in the proteolysis of Swiss cheeses, provided that they are released through bacterial lysis. Experimental Swiss cheeses were manufactured on a small scale from thermized and microfiltered milk using as starters (in addition to Streptococcus thermophilus and Propionibacterium freudenreichii) one of two Lb. helveticus strains, ITGLH1 and ITGLH77, which undergo lysis to different extents in vitro. All the cheeses were biochemically identical after pressing. The viability of Lb. helveticus ITGLH1 and ITGLH77 decreased to a similar extent (96–98%) while in the cold room, but the concomitant release of intracellular lactate dehydrogenase in cheeses made with strain ITGLH1 was 5–7-fold that in cheeses made with ITGLH77. Protein profiles and immunoblot detection of the dipeptidase PepD confirmed a greater degree of lysis of the ITGLH1 strain. Free active peptidases were detected in aqueous extracts of cheese for both strains, and proteolysis occurred principally in the warm room. Reversed-phase HPLC revealed a more extensive peptide hydrolysis for ITGLH1, which was confirmed by the greater release of free NH2 groups (+33%) and free amino acids (+75%) compared with ITGLH77. As the intracellular peptidase activities of ITGLH1 and ITGLH77 have previously been shown to be similar, our results indicated that the extent of lysis of Lb. helveticus could have a direct impact on the degree of proteolysis in Swiss cheeses.