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.

The effects of addition of EDTA on the dissociation of caseins and foaming properties of milks (100 g solids/l) reconstituted from skim milk powders given a low-heat (72°C for 30 s) or high-heat (85°C for 30 min) treatment during powder manufacture were determined. The EDTA-induced dissociation of caseins was independent of heat treatment but in high-heat milk was accompanied by release of denatured whey proteins. EDTA changed the proportions of individual caseins in the supernatant. EDTA addition improved both foam overrun and foam stability of low- and high-heat milks. The increase in serum protein on addition of EDTA contributed to the improvement in foaming properties of milks by increasing the availability of the proteins for formation of the air–water interface.

To obtain information about the possible release of β-casomorphins from β-casein under in vitro conditions, cows' milk was incubated with 13 strains of Gram-negative or Gram-positive bacterial species isolated from bovine milk. After incubation periods of 1–24 d, milk samples were assayed for β-casomorphin-4, -5, -6 or -7 immunoreactive materials. In general, no β-casomorphin immunoreactive material was found in samples incubated with non-caseolytic strains, e.g. Pseudomonas putida or Streptococcus faecalis subsp. faecalis; however, high levels were found in several samples incubated with caseolytic strains, e.g. Ps. aeruginosa or Bacillus cereus. When such materials were present, the amounts of β-casomorphin-4, -6 or -7 greatly exceeded β-casomorphin-5 immunoreactivity. It was concluded that while some of these materials probably represented authentic β-casomorphins, others were more likely to be β-casomorphin precursors.

Reconstituted skim milks (10 % w/w total solids, pH 6·7–8·0) were ultrasonicated (20, 400 or 1600 kHz at a specific energy input of 286 kJ/kg) at a bulk milk temperature of <30 °C. Application of ultrasound to milk at different pH altered the assembly of the casein micelle in milk, with greater effects at higher pH and lower frequency. Low frequency ultrasound caused greater disruption of casein micelles causing release of protein from the micellar to the serum phase than high frequency. The released protein re-associated to form aggregates of smaller size but with surface charge similar to the casein micelles in the original milk. Ultrasound may be used as a physical intervention to alter the size of the micelles and the partitioning of caseins between the micellar and serum phases in milk. The altered protein equilibria induced by ultrasound treatment may have potential for the development of milk with novel functionality.

Whey protein complexed and became co-sedimentable with casein micelles after heating milk at ≥ 90°C for 10 min at pH ≤ 6·9 while at higher pH values (7·3) whey proteins and κ-casein-rich protein dissociated from the micelles on heating. κ-Casein-deficient micelles were more sensitive to heat, Ca2+ or ethanol than whey protein-coated or native micelles and were readily coagulable by rennet. Isolated κ-casein added to skim milk before preheating (90°C for 10 min) did not associate with the micelles at pH ≥ 6·9. Sodium dodecyl sulphate increased the level of both non-sedimentable N (NSN) and N-acetylneuraminic acid (NANA) and shifted the NSN-pH and NANA-pH curves to more acidic values while cetyltrimethylammonium bromide had the opposite effect. It is suggested that the pH-dependent dissociation in micellar κ-casein, which appears to be reversible, depends on the surface charge on the micelles; at a certain negative charge, disruption of hydrophobic and electrostatic forces could result in the dissociation of κ-casein from the casein micelles.

Measurements of residual native protein remaining after heat treatment of cheese whey have been made using quantitative polyacrylamide gel electrophoresis. The results have been expressed in terms of kinetic constants. The effect of concentration was investigated up to 3 times the normal total solids content, dialysis treatments were used to study the effect of non-protein constituents, and the effect of pH was studied at pH 4, 6 and 9. The results indicated that increased total solids (TS) concentration slowed the denaturation of β-lactoglobulin A and B (β-lg A, β-lg B) but hastened the denaturation of α-lactalbumin (α-la). However, increased lactose concentration slowed the denaturation of both α-la and β-lg, perhaps by preventing formation of heat-induced complexes. Increased Ca concentration, up to 0.4 mg/ml, tended to slow the denaturation of both proteins, but further increase in Ca up to 0.9 mg/;ml produced little effect. The rate of denaturation of both α-la and β-lg was slower at pH 4 than at pH 6 or 9, and was probably slowest at the isoelectric point. However, not all the changes associated with pH could be explained in terms of net molecular electrostatic charge. The genetic variants of ′β-lg showed different heat stabilities – below 90 °C β-lgA was more stable than β-lgB, but above 90 °C the situation was reversed at all TS concentrations, and pH 6. However, at pH 4 and 9, °-lgA was less stable than °-lgB over the entire temperature range at normal concentration.

Mixed cultures of different EPS-producing Streptococcus thermophilus strains in combination with a Lactobacillus delbrueckii subsp. bulgaricus strain with negligible EPS-production were used for yoghurt production. The yoghurt texture was characterised with respect to sensory, rheological and microstructural properties and the EPS-concentrations were determined. The cultures resulted in yoghurts with highly different texture properties, and positive interactions between certain Streptococcus thermophilus strains were observed. The underlying properties of yoghurt texture are multidimensional, but a number of microstructural characteristics were apparent in the yoghurts with the highest mouth thickness, creaminess and viscosity. A strong protein network, not too dense and with medium size pores containing EPS, seems associated with these properties. The presence of capsular polysaccharides (CPS) also appeared to be beneficial as did a combination of EPS types, which were distributed differently in the protein network (in serum pores, respectively in association with protein). Obviously, a certain concentration of EPS must be present to provide for these effects on yoghurt texture, but other factors than concentration per se seem more important.

The present study describes the purification of a post-proline dipeptidyl aminopeptidase from the cytoplasm of Streptococcus cremoris AM2. On the basis of its elution from a calibrated Sephadex G200 column, the enzyme had a molecular weight of 117000 and exhibited a broad pH optimum activity between 6·0 and 9·0. The activity was most comprehensively inhibited by phenylmethylsulphonylfluoride and more modestly inhibited by 1,10-phenanthroline and 8-hydroxyquinoline but not by EDTA. A range of peptides containing either proline or alanine as the penultimate amino acid residue could act as substrates. The presence of proline on the carboxy side of the scissile bond prevented hydrolysis. However the enzyme could release Pro-Pro from Pro-Pro-Gly-Phe-Ser-Pro. The significance of this substrate specificity is considered in the context of removal of either single proline residues or prolylproline sequences from oligopeptides during cheese ripening.

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 novel 2-stage gravity separation scheme was developed for fractionation of raw, whole bovine milk into fractions enriched in small (SFG) or large (LFG) fat globules. The volume mean diameter of fat globules in SFG, LFG or control (CTRL) milk was 3·45, 4·68 and 3·58 μm, respectively. The maximum in storage modulus (index of firmness) decreased with increasing fat globule size for rennet-induced gels formed from SFG, LFG or CTRL milks. Miniature (20 g) Cheddar cheeses were manufactured using each of the 3 milks. There were no significant (P>0·05) differences in the pH, moisture and fat in dry matter levels between cheeses made using any of the 3 milks, however, the fat content of the cheese made using SFG milk was ~1% lower than that of cheese made using LFG or CTRL milk in each of the 2 trials. Image analysis of confocal scanning laser micrographs of the cheeses illustrated that the star volume of fat globules in the cheeses decreased significantly (P[les ]0·05) as the size of fat globules in the milks used for cheesemaking was reduced. This indicates that it is possible to manipulate the size distribution of fat globules in Cheddar cheese by adjusting the fat globule size distribution of the milk used for cheesemaking. The concentration of free fatty acids (FFA) increased in all cheeses during ripening. At 120 d of ripening, the concentration of FFA varied significantly (P[les ]0·05 and P[les ]0·001 for trials 1 and 2, respectively) with fat globule size, with cheeses made in trial 2 from LFG, SFG or CTRL milks having total FFA levels of 3391, 2820 and 2612 mg/kg cheese, respectively.

Heat sterilization of milk destroyed about 50% of its folic acid activity, and subsequent exposure of the milk to sunlight caused a progressive further loss. This loss of folate, as of vitamin B12, was a consequence of the oxidative destruction of the milk's ascorbic acid, and was largely prevented by thorough exclusion of oxygen from the milk during heat processing and subsequent storage. With thiamine, riboflavin, nicotinic acid, vitamin B6 and biotin, stability towards heating and during exposure to sunlight was affected little, if at all, by the oxygen tension in the milk.

In this study, the antihypertensive activity in spontaneously hypertensive rats of two peptides isolated from β-lactoglobulin hydrolysates with thermolysin was evaluated. These peptides, with sequences LLF [β-lg f(103–105)] and LQKW [β-lg f(58–61)], showed potent in vitro ACE-inhibitory activity. Two hours after administration, both sequences caused a clear and significant decrease in the blood pressure of these rats. The impact of a simulated gastrointestinal digestion on ACE-inhibitory and antihypertensive activities of these peptides was also studied. The results showed that both fragments were susceptible to proteolytic degradation after incubation with pepsin and Corolase PP®. In addition, their in vitro ACE-inhibitory activity decreased after the simulated digestion. It is likely that fragment LQK was the active end product of the gastrointestinal digestion of peptide LQKW. The fragment LL, observed after digestion of peptide LLF, probably exert its antihypertensive effect through a mechanism of action different than ACE-inhibition.

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.

Milk was heated at 97 °C/15 s to denature ∼ 30% of the whey protein and then used to make Cheshire cheese. Measurements of para-κ-casein production indicated that heating milk did not inhibit the enzymic action of rennet, but additional Ca and an initial pH of 6·4 were required for normal coagulation and curd-firming. In the experimental cheeses, about 4·5% more dry matter was recovered compared with controls made with pasteurized milk owing to a 6·7 and 0·7% increase in protein and fat recovery respectively. Experimental cheeses tended to be too moist and the curds did not fuse as well but these problems could be overcome by raising the scald temperature and cheddaring the curds. When the moisture in non-fat solids of control and experimental cheeses was similar (61%), the flavours and textures were not significantly different. The procedure requires little modification to existing commercial plant and should be suitable for varieties of cheese with higher moisture content and more crumbly texture than Cheddar.

1. A study was made of the milking of the two hind quarters of four cows by teat cup and by teat cannula at three levels of vacuum and by teat cannula at atmospheric pressure.

2. The eight quarters milked at significantly different maximum rates by teat cup but at nearly the same rate through a teat cannula, suggesting that the teat orifice is a very important factor controlling rate of milking.

3. The maximum rate of milking by both teat cup and teat cannula increased with increasing level of vacuum from 11 to 20 in. Hg. The rate of increase in the teat-cup milking was greater than in the cannula milking, suggesting that the teat orifice was stretched open at the higher levels of vacuum.

4. The average amount of strippings increased at the higher level of vacuum in teat-cup milking, due apparently to teat-cup crawl.

Numerous methods based on DNA analysis have been employed in the food industry to monitor adulterations of food products of animal origin. Among them the most frequently used are: polymerase chain reaction (PCR) amplification of a marker gene fragment(s) with universal primers, or amplification of DNA with species-specific primers. PCR-products of different origin can be discriminated by size, restriction fragment length polymorphism (RFLP) or single stranded conformational polymorphism (SSCP) analysis. These methods have been used for identification, and differentiation between, the animal origins of raw or heat-treated meat and meat products (Chikuni et al. 1994; Meyer et al. 1994, 1995; Zehner et al. 1998; Behrens et al. 1999; Guoli et al. 1999; Hopwood et al. 1999; Matsunaga et al. 1999; Wolf et al. 1999). These approaches are also applicable to the analysis of dairy products. However, adulterations of goats' milk and its products are traditionally tested by immunological and/or electrophoretic methods (Amigo et al. 1992; Levieux & Venien, 1994; Mimmo & Pagani, 1998). So far, only a few DNA-based techniques designed to detect the presence of bovine DNA in goats' milk have been described (Plath et al. 1997; Branciari et al. 2000). This paper presents a one-step PCR procedure for detection of adulteration of goats' milk with cows' milk. The method, employing bovine-specific primers for amplification of a 274 bp fragment of cytochrome b DNA, seems to be simple, fast, specific and sensitive.

The irreversible heat denaturation of α-lactalbumin (α-la) in 0·1 M-phosphate, pH 7·0, at 100 °C was studied using polyacrylamide-gel electrophoresis (PAGE). PAGE revealed two groups of bands, one moving faster than native α-la and one slower, in addition to some denatured protein which remained at the origin and some residual native α-la. The faster group had unchanged molecular weight, but an increase in charge, partly due to hydrolysis of glutamine and asparagine residues. The slower group was shown by two-dimensional sodium dodecyl sulphate-PAGE to be oligomers of denatured α-la; formation of the smaller oligomers preceded the larger ones. The oligomers reverted to monomers in the presence of dithiothreitol, showing that they were disulphide-linked aggregates of denatured α-la. Immuno-blots of the gels showed that both fast and slow groups of bands had irreversibly lost most of the antigenicity of the native protein.

In the first part of this study, brief notes are given on the better known varieties of the soft pickled group of cheese which the authors believe to be best suited to warm climatic conditions.

Domiati cheese, a variety of this group, characterized by the addition of an appreciable amount of salt to the milk prior to rennetting, is described in detail. An outline of the steps of its manufacture, the pickling process, and the effect of the amount of salt added to the milk on the composition of the resulting cheese is given.

In the second part the effects of varying additions of salt to both cows' and buffaloes' milk on rennet coagulation, rate of whey drainage, chemical composition and yield of cheese and whey were investigated.

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.