In each of 5 cows, 2 quarters which had been regularly milked throughout the whole of pregnancy produced less milk in the next lactation than the other 2 quarters which had a normal dry period. The concentrations of the individual constituents in the milk of all 4 quarters of each cow were similar. The effect appears to be due to factors located within the individual quarter of the mammary gland rather than to factors such as the hormonal background or plane of nutrition which affect the whole animal.

The susceptibility of total casein and the individual caseins in reconstituted skim milk to transglutaminase (TGase)-induced cross-linking was studied as a function of incubation temperature (5–40 °C), pH (5·0–7·0) and mineral addition. Within the ranges studied, the level of total casein cross-linked increased with increasing temperature, pH and concentration of added trisodium citrate, whereas adding calcium chloride had the opposite effect. These effects can be largely related to the effects of these parameters on TGase activity. In addition, the parameters were also found to influence the susceptibility of κ-casein, and to a lesser extent β-casein, to cross-linking, whereas the susceptibility of αs1-casein was not affected. The susceptibility of κ-casein to cross-linking increased with increasing temperature and calcium chloride addition, but decreased with increasing pH and citrate content, whereas the susceptibility of β-casein to TGase-induced cross-linking decreased with increasing temperature, but was not affected by other parameters. These findings highlight the fact that selection of environmental conditions during cross-linking can be applied to tailor the surface, and hence possibly colloidal stability, of casein micelles in TGase-treated milk.

In this study, the chemical composition, proteolysis and in vitro angiotensin-converting enzyme-(ACE)-inhibitory and antioxidant activities of white cheeses made using probiotic adjunct cultures (Bifidobacterium bifidum DSMZ 20456 and Lactobacillus acidophilus DSMZ 20079) were investigated. The cheeses were ripened in a vacuum package or brine for 120 d at 4 °C. The cheese samples maintained the probiotic characteristics of the viable cells as >106 cfu/g even after ripening for 120 d. The proteolysis degrees in terms of water-soluble nitrogen/total nitrogen (WSN/TN), trichloroacetic acid-soluble nitrogen/total nitrogen (TCA-SN/TN) and phosphotungstic acid-soluble nitrogen/total nitrogen (PTA-SN/TN) values in the cheeses increased throughout the ripening. The highest levels of proteolysis were found in cheese made using Lb. acidophilus DSMZ 20079 and ripened in a vacuum package. ACE-inhibitory activity of the water soluble extracts (WSEs) of the cheeses increased significantly (P < 0·05) throughout the ripening (IC50 values 82·78–140·99 μg/ml). Use of Lb. acidophilus DSMZ 20079 and packaging under vacuum significantly increased the percentage of ACE inhibiting activity. WSEs had DPPH scavenging activity (the IC50 values were 2·41–5·39 mg/ml and the inhibition values were 5·10–10·38%), increasing up to 60 d ripening. In the present study, it was observed that Lb. acidophilus DSMZ 20079 was more effective than Bifido. bifidum DSMZ 20456 in terms of the cheese characteristics investigated.

Kefir is a refreshing fermented milk with a slightly acidic taste obtained by incubating milk with kefir grains (Saloff-Coste, 1996). Kefir grains are a complex mixture of lactic acid bacteria and yeasts in a strong and specific association. They are characterized by an irregular form, a folded and uneven surface and a white or slightly yellow colour. They are tough and resilient and have a characteristic acid taste (Bottazzi et al. 1994). The basic microflora contains lactococci, homofermentative and heterofermentative lactobacilli, yeasts and acetic acid bacteria (Bottazzi et al. 1994; Rea et al. 1996). Among the yeasts isolated from grains and identified are Candida kefir, Saccharomyces cerevisiae, Sac. delbrueckii, Cand. holmii, Sac. unisporus and Sac. lipolytica (Angulo et al. 1993; Marshall, 1993; Garrote et al. 1997). Lactobacillus brevis, Lb. viridescens, Lb. casei, Lb. kefir, Lb. kefiranofaciens, Lb. kefirgranum, Lb. parakefir, Leuconostoc spp. and Lactococcus lactis are among the lactic acid bacteria present in the grains (Marshall et al. 1984; Toba et al. 1991; Takizawa et al. 1994; Garrote et al. 1997). The mixed microflora of yeasts and bacteria is held together by a matrix containing (g/kg dry weight) protein 340, polysaccharide 470 (Ottogalli et al. 1973).

The study of kefir grains in milk has been centred on the characteristics of the polysaccharide produced by lactobacilli within the grain (Yokoi et al. 1991). This polysaccharide, named kefiran, is composed of glucose and galactose (Yokoi et al. 1991). It has been suggested that proteins are incorporated from the growth media (Bassette & Acosta 1988), but no details about structure and composition are available.

Soyabeans are an important component of the diet in many countries and have been used to obtain fermented products such as sogurt (Mann, 1991). Special attention has been given to the growth of, and sugar utilization by, Bifidobacterium spp., Lb. delbrueckii subsp. bulgaricus and Streptococcus thermophilus in soya milk (Buono et al. 1990; Murti et al. 1993; Ankenman Granata & Morr, 1996). To our knowledge, there have been no attempts to ferment soya milk with kefir grains.

The aim of this study was to investigate the growth of kefir grains in soya milk and the composition of these grains, focusing on the matrix proteins.

(1) Increasing the pre-heating temperature of the milk from 165 to 190° F. before spray-drying greatly improved the resistance of the powder to the development of tallowy ‘off’-flavours on storage. From the results of accelerated tests at 47 and 37° C. a storage life (in air) of the order of 2 years could safely be predicted for the powders from milk pre-heated at the higher temperature. Confirmation has now been obtained by storage of the powders for 2½ years at 15° C. and at room temperature.

(2) Improvement in the bacteriological quality of the milk supply, and more careful control during its passage through the plant, produced some improvement in the keeping properties of powders prepared from milk pre-heated at the lower temperature, but had relatively little effect on the keeping properties of the powder when the higher preheating temperature was used.

(3) The shapes of the deterioration curves for the various powders showed that the effect of the higher pre-heating temperature and of improvement in the milk supply was to extend the induction period which precedes rapid deterioration. The protective effect of both factors was greater when measured at the ‘slightly tallowy’ or ‘fairly good’ stage than when comparison was made at a more advanced stage of decomposition.

Fifteen lactating cows were milked throughout pregnancy, and the effects on milk performance were studied during this period and during the succeeding lactation, relative to 11 conventionally managed cows (2 months dry before calving) as controls. During the last 2 months of pregnancy, only nine cows did not dry off spontaneously. Protein and fat concentrations in milk increased rapidly, but the concentration of lactose, corrected for milk yield, did not change. The ratios of individual caseins to total protein decreased with the quantity of milk produced, but only for yields below ˜ 6 kg/d. The relative proportion of κ-casein tended to decrease in the last milkings. During the succeeding lactation (first 15 weeks after calving and first 6 weeks of grazing) continuously milked cows yielded 4 kg milk/d less than the cows of the other group. The protein content of their milk was higher (2–3 g/kg depending on the period) than that of the control group, and the lactose content tended (p < 0·10) to be lower. Changes in the relative proportions of nitrogenous fractions with time were not different in the two groups. Differences between the two groups in the concentration of protein in milk, and in the concentration of glucose and non-esterified fatty acids in the plasma, suggest a better energy balance for the continuously milked cows during the succeeding lactation.

Proteolysis during storage of UHT skim and whole milks processed by either direct or indirect systems has been studied. All the proteolysis indices determined (measurement of free amino groups, PAGE of caseins and PAGE and reversed-phase HPLC of the fraction soluble at pH 4·6) revealed greater proteolytic degradation during storage of skim milks compared with that of whole milks subjected to the same UHT treatments. Increased activities of both native milk proteinase and proteinases of bacterial origin were observed in skim UHT milks. The different behaviour of UHT skim and whole milks on storage would have to be taken into account in establishing the process conditions.

The effect of homogenization on the heat stability characteristics of milk was examined. The heat stability of homogenized milk, as determined by the time taken for protein clots to form when heated at 140 °C, was reduced with increasing pressure in the range 3·5–34·5 MPa. The heat stability of homogenized milks was greater for samples obtained in the summer months than for those obtained in the winter. The general destabilizing effect of homogenization could be partly offset by 2-stage homogenization (20·7 MPa followed by 3·5 MPa), addition of phosphate stabilizers (0·08% w/v) or homogenization at a high temperature (65 °C). Whilst homogenized and unhomogenized milks reacted similarly to the addition of Ca, phosphate stabilizers, sulphydryl-blocking and oxidizing agents, the effects of season, addition of urea or formaldehyde were different for homogenized milk.

The heating of milk causes denaturation of the lactalbumin and lactoglobulin to an extent depending on both the temperature and the time of heating.

A series of data has been given for the amounts of albumin and globulin denatured on heating samples of milk at various temperatures from 63 to 80° C, for periods of from 2½ to 60 min.

An average of 10·4 per cent, of the total soluble protein was denatured after 30 min. heating at as low a temperature as 63° C.

Smooth curves were obtained for the progress of denaturation with time at each temperature, and, over the range of 63–75° C, the relative increase in velocity of denaturation for each rise in temperature of 1° C. was found to be constant, the temperature coefficient of the reaction being 1·5.

A method for obtaining distillates from whole Cheddar cheese is described. The distillate has an aroma at −80°C closely resembling that of the cheese before distillation. Gas chromatograms of the distillate were obtained by sampling the vapour over the distillate at −80°C, using both a 15-m Apiezon surface-coated open tubular column and a 1·5-m glass column packed with Porapak Q. The aroma of the effluent was evaluated and the chromatograms simultaneously recorded by passing the vapour through a splitting device before the flame-ionization detector of the gas chromatograph. The compounds responsible for the aromas detected in the effluent were identified using a gas chromatograph directly linked to a mass spectrometer. Of the compounds identified in the distillate, H2S, methanethiol, dimethyl sulphide and diacetyl had the most intense aromas, and these compounds are discussed in relation to their importance to Cheddar-cheese aroma.

The stability of homogenized ultra-heat-treated (UHT) aseptically-packed cream of 18 % fat content when added to hot coffee was indirectly monitored in 10 batches of cream, stored for 10 weeks at different temperatures. A tendency to coagulate under standard conditions (instability) is known as feathering. Stability was determined by a procedure in which acetate buffer instead of coffee solution was used as the test solution. The distribution of components between the fat and aqueous phases of the stored cream was also monitored. Feathering score (stability) decreased during storage but was not accompanied by any increase in non-protein nitrogen levels. The amount of casein and Ca in the fat phase of the cream increased with time and was inversely related to feathering score. Some batches of cream contained 0·1 % Na2CO3 and 0·1 %Na citrate as stabilizers and these additions were found to improve feathering scores early in the storage period. Initially, the fat phase of these creams contained less casein and Ca than those without the additives, but after 10 weeks the differences in feathering score and in casein and Ca distribution, between the 2 types of cream diminished. Examination by electron microscopy showed that casein micelles were associated with fat globules, apparently linking several globules together, which was less in the creams containing additives, but after 10 weeks the appearance of all creams was similar. Electrophoretic separation of the fat globule membrane (FGM) proteins by SDS-polyacrylamide-gel electrophoresis showed that casein was the principal protein component, but that β-lactoglobulin and native FGM proteins were aslo present.

1. The effect of feeding levels of protein and energy appreciably higher than the Woodman standards on the yield and composition of the milk from twelve Ayrshire cows has been studied in two winter-feeding experiments.

2. In the first experiment the S.E. of the concentrate ration was held constant at 63½% and the D.C.P. content adjusted to 10, 15 and 17%. The 10 and 15% D.C.P. concentrates derived their protein from home-grown feedingstuffs, whereas the 17% D.C.P. concentrate was a bought-in commercial mixture. Feeding the three levels of D.C.P. led to no statistically significant differences in milk yield, milk composition or live-weight gain, although the commercial mixture (17% D.C.P.) resulted in milk with a slightly lower fat content.

3. The second experiment reversed the feeding conditions of the first, i.e. D.C.P. was held constant at 16% and three concentrates were fed which contained 59, 67 and 75% S.E. respectively. The two rations of higher S.E. content gave significantly more milk than the low S.E. ration and tended to give slight increases in live weight. There was a slight increase in the S.N.F. content of the milk when the S.E. of the concentrate was raised from 59 to 67% S.E. This was due almost entirely to an increase in the protein content of the milk. There was no further increase in S.N.F. with the 75% S.E. concentrate, and this latter concentrate produced milk with a slightly lower fat content.

4. Comparison of costs and returns showed that the cheap low protein concentrate, ration C, even though it gave the least milk, yielded the highest profit. It was shown, however, that at any reasonable fixed cost of starch equivalent the most profitable level of feeding above the Woodman maintenance standard was 3–3·3 lb. S.E./10 lb. of milk of 3·8–4·0% fat. Alternatively, the total ration should be 110–117% of the Woodman standard.

Udder defence mechanisms are not completely explained by current mastitis research. The anatomical construction of the udder implies that infection of one udder quarter does not influence the immune status of neighbouring quarters. To test this hypothesis, we compared the immune reactions of individual udder quarters in response to microbial attacks. In the course of immune reactions, polymorphonuclear leucocytes (PMN) release oxygen radicals, which can be determined by chemiluminescence (CL). Milk from 140 udder quarters of 36 cows was analysed for somatic cell count (SCC), differential cell count, viability and CL activity. Quarters with an SCC <100000 cells/ml and free of pathogens were defined as uninfected, all other quarters were categorized as infected. Three groups of cows were classified cytologically: group A (healthy, 11 animals, SCC limit <100000 cells/ml); group B (moderate mastitis, 8 cows, SCC [ges ]100000 and <400000 cells/ml in at least one quarter); and group C (severe mastitis, 17 cows, SCC [ges ]400000 cells/ml in at least one quarter). Infected and uninfected quarters in groups B and C were analysed separately. Viability of PMN leucocytes was significantly (P=0·0012) lower in group A (72·6%) than in healthy quarters of group C (84·0%). Lowering the SCC limit of healthy quarters to <50000 cells/ml (group A: all quarters within the udder) revealed striking differences between samples of groups B and C: in addition to varying differential cell counts and viabilities, CL activity of group B<50 (2929 CL units/million PMN) was markedly lower than that of the other groups (5616 in group A<50 and 6445 CL units/million PMN in group C<50). These results allow the conclusion that the infection of one udder quarter influences the cell activity of neighbouring quarters. When the SCC threshold for healthy quarters was reduced to 50000 cells/ml, greater differences in cell activities were detected between healthy udders and healthy quarters of infected udders.

1. Four test organisms were used for the microbiological assay of vitamin B6. The test organisms and the forms of vitamin B6 active for them were: Saccharomyces carlsbergensis 4228 (pyridoxine, pyridoxamine and pyridoxal), Streptococcus faecalis R ATCC8043 and Str. faecium Ø51 (pyridoxamine and pyridoxal), and Lactobacillus casei ATCC7469 (pyridoxal).

2. By simultaneous assay of the test material with the four organisms the amounts of pyridoxine, pyridoxamine and pyridoxal present were determined. The differential assay technique has been improved by the use of Str. faecium, which is more specific for measuring total pyridoxamine plus pyridoxal, instead of Str. faecalis.

3. The optimum conditions for measuring vitamin B6 in milk were studied and the differential assay technique was applied in an investigation of the effect of heat on the vitamin B6 activity of milk.

4. It was found that pyridoxal accounted for about 80% of the vitamin B6 activity of raw milk, the other 20% being due to pyridoxamine.

5. In the manufacture of evaporated milk there was a marked fall in the pyridoxal and a slight increase in the pyridoxamine content. The overall loss was some 60%.

6. On storage of evaporated and of sterilized liquid milk at room temperature there was a further loss of vitamin B6 activity. Most of the residual activity in stored evaporated milk appeared to be due to a substance other than pyridoxal or pyridoxamine, possibly pyridoxine.

The components of whole casein from water buffalo (Bubalis L.) have been re-investigated in the light of recent studies elucidating the primary structure of all the bovine caseins (αs1, αs2, β & κ). Whole casein samples obtained from individual Italian buffaloes all contained 2 αs1- and 2αs2-casein fractions. These 4 components were isolated and identified by their amino acid compositions and terminal amino acid sequences. The 2 αs1-casein fractions seem to have identical peptide chains as do the 2 αs2 fractions, with the individual fractions within each group differing only in their phosphate content. Buffalo β-casein was also isolated and characterized by its amino acid composition and terminal sequences. The isolation of different buffalo κ-casein fractions was reported earlier (Addeo, Chobert & Ribadeau-Dumas, 1977). These fractions were purified and analysed. They were found to have the same amino acid compositions and C-terminal sequences, but to differ in their carbohydrate content. The prominent component, κ1, was treated with chymosin; the caseinomacropeptide and para κ-casein were separated from the digest and analysed. Starchgel electrophoresis at alkaline pH showed no polymorphism of αs1-, β- and κ-caseins in 170 individual buffalo whole casein samples. On 5 occasions the electrophoretic pattern of αs2-casein suggested genetic polymorphism. The behaviour of buffalo αs1−and αs2-caseins in the presence of Ca2+ was also studied.

Milk somatic cell count (SCC) is the gold standard in diagnosis of subclinical mastitis, and is also an important parameter in quality programmes of dairy cooperatives. As routine SCC analysis is usually restricted to central laboratories, much effort has been invested in the search for alternative biomarkers of mastitis and milk quality, including the presence in the milk of the acute phase proteins (APP), haptoglobin (Hp) and serum amyloid A (SAA). The aim of this study was to investigate relationships between Hp, SAA and SCC in quarter, cow composite, and bulk tank milk samples. Cows (n=165), without any clinical signs of disease or abnormalities in the milk or udder, from three different dairy farms, were used. Cow composite milk samples from all cows delivering milk at the sampling occasion were taken once in each herd. In one of the farms, representative quarter milk samples (n=103) from 26 cows were also collected. In addition, bulk tank milk samples from 96 dairy farms were included in the study. Samples were analysed for Hp, SAA and SCC, and relationships between the parameters were evaluated at quarter, cow and tank milk levels using Chi-square analysis. Milk samples were categorized according to their SCC, and the presence, or no presence, of SAA and Hp, based on the detection limits of the screening methods (0·3 mg/l and 1·0 mg/l for SAA and Hp, respectively). Hp and SAA were found in milk at quarter, cow composite and bulk tank levels. A large proportion (53%) of the animals had detectable milk concentrations of APP, and SAA was detected more frequently, and at higher concentrations than Hp, regardless of sample type. SAA was detected in as many as 82% of the bulk tank milk samples. Significant relationships were found between Hp, SAA and SCC at quarter and cow composite milk levels, but only between SAA and SCC at bulk tank milk level. Detectable levels of APP were more common at high SCC.

Casein micelles in milk-salts solution adsorbed charged detergents and highly-charged polypeptides strongly, neutral detergents less strongly and low molecular-weight amines weakly. A tetra-amine was adsorbed more strongly than a tri-amine. The extent of adsorption of proteins tended to rise as the molecular weight increased. Glycerol and lactate were adsorbed to a limited extent but dextran and α-ketoglutarate were not adsorbed at all. Proline was partly adsorbed, indicating that hydrophobic binding sites were available, and caused some disruption of the casein micelles. Additives were bound to approximately the same extent by casein micelles and rennet coagula. The proportions adsorbed were constant over at least 10-fold ranges of concentration. Additives which increased the rennet clotting time (RCT) acted by binding Ca2+. Most additives decreased the RCT, the extent increasing with the amount adsorbed and the positive charge on the additive. The greatest reduction in RCT was observed with those additives which had positively-charged and hydrophobic moieties and bound most strongly to casein micelles. Of the additives tested, only sodium dodecyl sulphate affected the enzymic action of rennet. The reduction in RCT may have resulted from the neutralization of the negative charge of the micelles or enhancement of their hydrophobicity, favouring hydrophobic interactions between the particles.

Skim milks were prepared from skim milk powder at several concentrations between 10 and 25% total solids and portions were pH-adjusted to between pH 6·3 and 7·1 and heated at 120°C. After ultracentrifugation (88000 g for 90 min), the supernatants were analysed using gel electrophoresis to determine the concentrations of β-lactoglobulin, α-lactalbumin and κ-casein. The dissociation of κ-casein from the micelles was dependent on both the pH and the total solids content of milk before heating. Both higher pH (in the range 6·5–7·l) and higher concentration increased the extent of dissociation. A further series of samples were heated for 2–11 min at 120°C at pH 6·55. κ-Casein dissociation increased with concentration and with heating time. It was concluded that as the milk increased in concentration, the pH at which micellar κ-casein dissociated on heating was lowered.

The overall objective of this study was to evaluate agreement between rectal (RT) and vaginal temperature (VT) measured with the same temperature loggers in dairy cows. Three experiments were conducted. The study began with a validation in vitro of 24 temperature loggers comparing them to a calibrated liquid-in-glass thermometer as a reference method. The association and agreement between the 24 temperature loggers with the reference method was r=0·996 (P<0·001) with a negligible coefficient of variance (0·005) between the loggers. In-vivo temperature loggers were tested in 11 healthy post-partum cows (Experiment 2) and 12 early post-partum cows with greater body temperature (Experiment 3). Temperature loggers were set to record VT and RT at 1-min intervals. To prevent rectal and vaginal straining and potential expulsion of temperature logger an epidural injection of 2·5 ml of 2% Procain was administered. Association between RT and VT was r=0·92 (P<0·001; Experiment 2) and r=0·94 (P<0·001; Experiment 3) with a negligible difference of −0·1 and 0·01 °C. Bland-Altman plots demonstrated agreement between RT and VT for healthy and early post-partum cows with greater body temperature in Experiments 2 and 3, respectively. Furthermore the intra-class correlation coefficient between RT and VT measured with identical loggers within cows of Experiments 2 and 3 also demonstrated greater agreements (P<0·001). Therefore, continuous VT monitoring with temperature loggers can be used as a measure of body temperature in dairy cows.

Using chromatographic techniques, a number of compounds was isolated from the steam distillates of skim milk containing oxidized (cardboard) flavour. The following compounds were definitely identified: acetone, acetaldehyde, n–hexanal, crotonaldehyde, and the C5 to C11 2-unsaturated aldehydes. Presumptive evidence for the presence of several 2,4-diunsaturated aldehydes of medium chain length was obtained also. Acetaldehyde and acetone were also isolated from normal skim milk; these two compounds appear to play no part in the defect.

2-Enals, in particular the C8 and C9, were considered to be the principal flavour-determining compounds. In milk at dilutions of 1 part in 107 to 109 they impart an oxidized type of flavour resembling that known as cardboard. Evidence was obtained that 2,4-dienals were also present and they produced a similar flavour.

It may further be deduced that cardboard flavour in milk is caused by the preferential oxidation of di- and polyunsaturated fatty acids.