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.

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. 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.

The experiments reported in this Research Paper aimed to study the effects of mustard and cumin seeds inclusion on feed utilisation, milk production, composition, and profile of milk fatty acids in lactating Damascus goats. The study was a completely randomised design with repeated measurements in time using fifteen goats (6 ± 2 d of lactation) divided into 3 treatments (5 goats per treatment). Goats were offered a control diet of berseem clover and concentrates (1 : 1 dry matter (DM) basis) or the control diet supplemented with either 10 g/d of dried mustard seeds (mustard treatment) or 10 g/d of cumin seeds (cumin treatment) for 12 weeks. Treatments had no effect (P > 0·05) on feed intake, but enhanced (P < 0·05) digestibility of DM, organic matter, non-structural carbohydrates, and fibre fractions. Digestibility was greater (P < 0·001) with cumin treatment compared with mustard treatment. Mustard and cumin seeds had greater (P < 0·05) ruminal total short chain fatty acids (SCFA) production, and molar proportion of propionate, with greater (P < 0·001) SCFA production for cumin vs. mustard treatment. Mustard and cumin seeds increased (P < 0·05) concentrations of serum total proteins, globulin, and glucose and lowered (P < 0·05) serum cholesterol concentration. Mustard and cumin seeds inclusion elevated milk production (P = 0·007), while cumin increased milk contents (P < 0·05) of fat and lactose. Cumin treatment lowered (P < 0·05) milk saturated fatty acids (SFA) and had greater total unsaturated fatty acids (UFA) and total conjugated linoleic acid (CLA) contents. Overall, supplementing diets of Damascus goats with mustard or cumin seeds at 10 g/daily enhanced feed digestion, ruminal fermentation, milk yield (actual production by 6·8 and 11·1%, and energy corrected milk yield by 10·1 and 15·4%, respectively) and positively modified milk fatty acid profile with a 3·9% decrease in milk individual and total SFA, and an increase in individual and total UFA by about 9·7%, and total CLA by about 23·1%.

The primary objective of this study is to validate a new fast method for determination of uric acid in milk. The method is based on an enzymatic-fluorometric technique that requires minimal pre-treatment of milk samples. The present determination of uric acid is based on the enzymatic oxidation of uric acid to 5-hydroxyisourate via uricase where the liberated hydrogen peroxide reacts with 10-acetyl-3,7-dihydroxyphenoxazine via peroxidase and the fluorescent product, resorufin, is measured fluorometrically. Fresh composite milk samples (n=1,072) were collected from both Jersey (n=38) and Danish Holstein (n=106) cows from one local herd. The average inter- and intra-assay variations were 7·1% and 3·0%, respectively. Percent recovery averaged 103·4, 107·0 and 107·5% for samples spiked with 20, 40 or 60 μm of standard, respectively, with a correlation (r=0·98; P<0·001) observed between the observed and expected uric acid concentrations. A positive correlation (r=0·96; P<0·001) was observed between uric acid concentrations using the present method and a reference assay. Storage at 4°C for 24 h resulted in lower (P<0·01) uric acid concentrations in milk when compared with no storage or samples stored at −18°C for 24 h. Addition of either allopurinol (a xanthine oxidase inhibitor) or dimethylsulfoxide (a solvent for allopurinol) did not affect milk uric acid concentrations (P=0·96) and may indicate that heat treatment before storage and analysis was sufficient to degrade xanthine oxidase activity in milk. No relationship was observed between milk uric acid and milk yield and milk components. Authors recommend a single heat treatment (82°C for 10 min) followed by either an immediate analysis of fresh milk samples or storage at −18°C until further analysis.

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 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.

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.

Cheddar cheeses were made from milks concentrated 3- to 6-fold by ultrafiltration. Modifications to the conventional cheesemaking process were designed to obtain the correct composition and pH value in the final curd, improve the fat and moisture retention and cheese texture, and increase the ripening rate. The compositions of all cheeses were similar to those of controls made with unconcentrated pasteurized milk. The curd and cheese structures and the textural properties of the cheeses were closer to those of the controls when made from milk concentrated 5-fold than when made from milk concentrated 3-fold. Light homogenization of the milk before concentration led to improved fat recovery and decreased graininess and crumbliness in the cheese. Lowering the coagulation temperature of concentrated milks gave improved curd structure and less grainy and more sticky cheeses. Addition of the bacterial proteinase, Neutrase, with the salt, stimulated proteolysis and flavour production in the cheeses and made them more sticky. Neither reduction of the mineral levels in the milk by acidification before concentration nor reduction of the lactose concentration by diafiltration had any significant effect on the composition or textural properties of the cheeses.

Concentration of milk by reverse osmosis is severely hindered by the formation of deposits on the membrane. Electron micrographs of this deposit show a thin (11nm) continuous electron-dense layer adjacent to the membrane. An electronlucent zone (10–15nm thick) separates this layer from the main bulk of the deposit which is 30 μm thick and consists largely of granules with some vesicles in a diffuse matrix. The density of packing of the granules increases with proximity to the membrane. Their appearance, size and digestion with pronase suggest that these are milk protein granules. The order and nature of these various layers could explain the pattern of flux decline.

The type and relative amounts of plasminogen activator (PA) in different fractions of bovine milk obtained from 15 Holstein cows were examined. Raw milk was centrifuged to separate skim milk and a somatic cell pellet. PA was mainly localized within the casein fraction, being 42 times that in the serum, and in association with somatic cells. The predominant form of PA in milk casein was isolated from SDS-PAGE gel extracts and had a molecular mass of ∽75 kDa. Its activity was increased 41-fold (P < 0·01) in the presence of fibrin but was unaffected by the presence of amiloride, indicating that it was due to tissue-PA. The predominant forms of PA associated with milk somatic cells were isolated from SDS-PAGE gel extracts and had molecular masses of ∽ 30 and ∽ 50 kDa. The activity of both proteins was unaffected by the presence of fibrin but was dramatically reduced by the presence of amiloride, indicating that they represented urokinase-PA.

A dynamic mathematical model of blood flow regulation in the mammary glands of a lactating cow was constructed from a principle of local vasodilator release in response to changes in intracellular adenylate charge. An equation was derived to predict uptake of the milk precursors acetate, glucose, amino acids and fatty acids, as affected by mammary blood flow rate. Metabolism of the precursors to milk components and CO2 was simulated with a set of empirically derived equations. Relative rates of ATP production and utilization regulated both the number of perfused capillaries and the conductance of arteriolar segments in the mammary glands. The model simulated local control phenomena of functional and reactive hyperaemia, and simulation of autoregulation under changing arterial pressure suggested a predominance of precapillary sphincter regulation. It was predicted that an increase in blood flow without the mammary capacity to utilize blood metabolites efficiently would be detrimental to milk synthesis. Conversely, increased blood flow through changes in mammary activity resulted in predictions of higher milk production. It was proposed that the equation for uptake,

be used in analysis of mammary arteriovenous differences.

A total of 120 milk samples were collected from Comisana ewes throughout lactation. The ewes were ranked into two somatic cell count (SCC) categories: normal milk (N Milk) with SCC lower than 5·00×105/ml and high somatic cell milk (HSC Milk) with SCC higher than 1·00×106/ml. Milk samples were analysed in triplicate for pH, fat and protein contents, renneting parameters, and plasmin and plasminogen activities. The peptide profile due to total proteolytic activity (endogenous and exogenous enzymes) on α- and β-CNs were determined using urea-PAGE on sodium caseinate (pH 8·0 and pH 5·0) incubated at 37 °C for 4 d after sampling. The peptide profile due to non-plasmin enzyme activities at pH 5·0 was also determined using urea-PAGE. Plasmin activity was higher in the HSC milk than in the N milk throughout the study period. A decrease in plasmin activity was observed in the N milk during mid-lactation, which was probably related to decrease in pH, and in the HSC milk during late lactation, which may be ascribed to an enhanced influx of plasmin inhibitors from the blood stream. Proteolytic patterns in Comisana ewe milk were mainly affected by plasmin activity that increased with the SCC in milk. Also non-plasmin proteolytic activity was strongly enhanced by elevated SCC and resulted in a higher degradation of α-casein than of β-casein. In general, plasmin activity did not increase with the advancement of lactation and exhibited a different trend in HSC and N milk, suggesting that physiological factors did not play a key role in regulating the plasminogen-plasmin system in ewes' milk. Plasmin activity, detected with the colorimetric assay was consistent with proteolytic activity on sodium caseinate shown in urea-PAGE electrophoregram.

Caseins (αs1, β, αs2, e κ) represent about 80% of the whole protein content of ruminant milk. Each of these proteins is encoded by single copy genes (CSN1S1, CSN2, CSN1S2 and CSN3, respectively) clustered on a ∼200-kb segment of chromosome 6 (Ferretti et al. 1990; Gallagher et al. 1994) in the order: CSN1S1, CSN2, CSN1S2 and CSN3 (Mercier & Vilotte, 1993). Furthermore, in cattle and goat CSN1S1 and CSN2 are convergently transcribed (Leroux & Martin, 1996; Rijnkles et al. 1997) and are only 20 and 12 kb apart, respectively.

Mastitis is the most frequent and costly disease in dairy herds, as it negatively affects yield and milk quality. The presence of clinical mastitis is quite easy to asses, whereas the diagnosis of the subclinical form can be more difficult and requires laboratory assays. Somatic cell count (SCC) is widely used as a rapid and low-cost indicator of mastitis, even if is not useful in discriminating between the clinical and subclinical form. As amyloid A has been investigated as a marker of mastitis, the aim of this study was to assess the potential value of measuring amyloid A in serum and milk and the correlation with SCC in the diagnosis of subclinical mastitis. The reliability of two different ELISA kits for the measurement of amyloid A in milk was also tested. During a 1-month trial period, 21 cows were assigned to three experimental groups according to their health status: 6 cows with clinical mastitis (CM), 10 cows with subclinical mastitis (SM) and 5 healthy cows (HE). Amyloid A was measured both in serum (SAA) and in quarter milk samples (mAA) with a serum ELISA kit, and in quarter milk samples (MAA) with a milk ELISA kit. SCC, total microbial count (TMC) and bacterial examination of the milk were also carried out. After a log transformation, the data were submitted to ANOVA and linear regression. TMC was significantly higher in cows with clinical mastitis, while no differences were observed between the other two experimental groups. SCC and MAA levels were significantly different among the three groups. mAA concentrations were similar between cows with subclinical and clinical mastitis, and SAA was not affected by mastitis. A significant correlation between SCC and MAA or mAA was detected, while no correlation was recorded between SAA and mAA. A close relationship between MAA and mAA was noticeable even at low concentrations, suggesting MAA as a potential physiological marker of subclinical mastitis.

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.

In this work the purification and the complete primary structure of κ-casein from equine milk are reported for the first time. Mares' milk casein was separated by RP-HPLC into four fractions. Complete primary sequence was obtained by sequence analysis of the protein in the fastest eluting peak isolated by chromatography. This sequence was 95% identical to that reported for the C-terminal portion of the zebras' κ-casein and showed high similarity with κ-caseins from sources other than Equidae, confirming that this protein was indeed κ-casein in equine milk. The presence of post-translational modifications in equine κ-casein was investigated by mass spectroscopy, after enzymic dephosphorylation. Two main components were found, the smaller component being more abundant. Equine κ-casein was recognized by a lectin specific for one of the glucosidic bonds in the saccharide moiety of bovine κ-casein. Sequence comparison with prevision studies showed that the distribution of charged and hydrophobic regions in equine κ-casein was similar, but not identical, to that found in the bovine protein; these regions are associated with the role of κ-casein in the formation and stabilization of the micellar structure of casein in milk.