Milk samples of four species of wild equids (onager, Equus hemionus onager; mountain zebra, E. zebra hartmannae; plains zebra, E. burchelli; Przewalski horse, E. caballus przewalskii) and two domesticated equids (ass, E. asinus; pony, E. caballus) were analysed. At mid to late lactation the milks of all species were very similar, containing on average 10–12% total solids, 1–2% fat, 1·6–1·8% true protein, 6–7% ‘lactose’, 0·3–0·5% ash, 0·08–0·12% calcium, 0·04–0·07% phosphorus and a calculated energy content of 2·0–2·4 kJ/g. Milk samples collected in the first 2 weeks after birth showed elevated levels of total solids and protein, and some had reduced ‘lactose’ levels, but there were no observable trends in milk composition during mid to late lactation (1–12 months post partum). It was concluded that these closely related species produce milks that are nearly identical in gross composition and that the domestic horse is a representative model for the study of equid lactation.

Mammary gland protein metabolism, determined by an arteriovenous difference technique, was monitored in four Holstein-Friesian dairy cows in response to supplemental dietary protein (provided as rumen-protected soyabean meal) during late lactation (weeks 24–30). Each cow was offered two isoenergetic diets composed of grass silage (170 g crude protein/kg dry matter) plus either a low (108 g/kg) or medium (151 g/kg) crude protein concentrate in a single crossover design involving two 21 d periods. On day 21, arteriovenous measurements across the mammary gland were made during a 13 h continuous i.v. infusion of [1-13C]leucine and with frequent (2 hourly) milk sampling during the final 6 h. Although total milk yield was slightly increased (+1 kg/d) by protein supplementation, milk protein yield was not significantly affected. Whole body protein flux (protein synthesis plus oxidation) was not significantly affected by supplementation. Total mammary gland protein synthesis (milk plus non-milk protein) was also not affected by supplementation but on both diets gland synthesis was always greater (by 20–59%) than milk protein output. The fractional oxidation rate of leucine by the mammary gland was significantly increased by protein supplementation (0·047 v. 0·136). Although the enrichment of leucine in secreted milk protein continued to increase, the final value (at 13 h) was 0·94 of the arterial plasma free leucine plateau value (not significantly different), suggesting almost exclusive use of plasma free leucine for milk protein synthesis. Based on current feeding schemes for dairy cattle, a fixed proportion (0·65–0·75) of the additional protein intake (+490 g/d) should have been partitioned into milk protein. Instead, leucine oxidation by the mammary gland was increased. Whether oxidation of other amino acids was also enhanced is unknown but if amino acid oxidation and the ‘additional’ non-milk protein synthesis occurring in the gland are not crucial to milk synthesis, then by reducing such activities improvements in the efficiency of converting absorbed amino acid into milk protein can be achieved.

Investigation of the effect of pH on the formation of deposit from milk during ultra high temperature treatment using a plate-type plant showed that deposit formation was greatly increased when the pH of whole milk was reduced to 6·54, irrespective of whether the adjustment was made through the addition of HCl or lactic acid. Most of the increase in deposition took place in the higher temperature sections of the plant. Conversely, an increase in milk pH to 6·8 using NaOH resulted in considerably less deposit being formed during heat treatment. Reducing the pH of whole milk increased the deposition of both protein and fat, but reduced the deposition of minerals. Despite very high concentration of fat in the deposits, it is unlikely that fat per se was responsible for increased deposit formation. Deposition also increased when the pH of skim milk was reduced to 6·51 before processing. Electron micrographs of the milks after heat treatment indicated that pH reductions caused the formation of large aggregates containing casein micelles during heating. Fat globules were also present in aggregates formed in whole milk with reduced pH. Slight reductions in the pH of milk before processing appear to enable the pH during heat treatment to fall below a critical value at which coagulation of milk takes place at the heated surfaces.

The heat denaturation of α-lactalbumin (α-la) in NaCl and KCl solutions, milk ultrafiltrate and milk was studied using the method of micro complement fixation. It was established that this protein was very resistant to heat denaturation and that it was more stable in milk ultrafiltrate than in the other media studied at temperatures up to 70 °C. Of the various milk proteins added to α-la, only β-lactoglobulin (β-lg) formed a heat-induced complex with this protein. This complex was identical in milk ultrafiltrate or in milk and depended on the molar ratio between both proteins; it was not modified by any other milk proteins. The binding of a-la to β-lg changed the ability of the latter protein to bind κ-casein.

A progressive change takes place in the heat stability of skim-milk during concentration. At the maximum in the coagulation time (CT)–pH profile of milk concentrated to over 20% of total solids (TS) the total N depletion curve changed from single- to 2-stage and CT became insensitive to the addition of urea. Furthermore, addition of β-lactoglobulin to skim-milk concentrates destabilized the heated milk whilst the opposite effect was observed in the presence of sulphydryl-group blocking agents. As a result of these observations, it has been suggested that the mechanism of coagulation in concentrated milk is similar to that which occurs within the minimum of the CT–pH profile of skim-milk at normal levels of TS.

Several microbial interactions involving yeast and lactobacilli have been suggested in fermented products. Co-aggregation between Lactobacillus kefir and yeast Saccharomyces lipolytica isolated from kefir grains was studied by scanning electron microscopy and aggregation assays. Six out of twenty Lb. kefir strains were able to co-aggregate with Sacch. lipolytica CIDCA 812 and showed thermolabile non-covalently bound surface molecules involved in this interaction. Co-aggregation inhibition after Lb. kefir pre-treatment with 5 m-LiCl or 20 g SDS/l showed that bacterial S-layer proteins play an important role in this interaction. Presence of different sugar (mannose, sucrose and fructose) or yeast pre-treatment with sodium periodate inhibited co-aggregation between Lb. kefir and Sacch. lipolytica. Co-aggregating Lb. kefir strains were also able to agglutinate with human red blood cells and they lost this ability after treatment with 5 m-LiCl. These results and the capacity of purified S-layer proteins of Lb. kefir to haemagglutinate, strongly suggest that a lectin-like activity of bacterial surface proteins (S-layer) mediates the aggregation with yeast cells.

Large differences in new infection rates occurred with half-udder milking machines in which one half of the cluster operated substantially free from vacuum fluctuations while the other half was subjected to various combinations of different types of fluctuation in vacuum. To increase the rate of new infection all teats were contaminated before and after each milking by immersion in a suspension of Streptococcus agalactiae and Str. dysgalactiae. A series of short-duration trials indicated that neither irregular nor cyclic fluctuations in vacuum acting alone were able to increase new infection rate. However, either the Nyhan and Cowhig irregular fluctuation, or a variant of it in combination with cyclic fluctuations in vacuum, was able to do so under the particular experimental conditions used.

Casein fractions and their breakdown products in Iberico-type cheeses made from the milk of cows, ewes or goats were analysed by capillary electrophoresis in order to characterize them. The actions of plasmin and chymosin on caseins were evaluated by comparing the electropherograms of caseins from milk and from cheese, both with and without treatment with plasmin. Characteristic capillary electrophoresis patterns were obtained for cheeses made from the milk of each of the three species, and the main components were identified. Caprine para-κ-casein and bovine β-caseins, eluting at the first and at the last part of the electropherogram respectively, were found to be indicative of the presence of the milks of these species.

The pH and calcium activity of reconstituted skim milk solutions (9–21% w/w milk solids non-fat) on heating and after cooling were studied as a function of milk pH prior to heating (pH 6·2–7·2 at 25°C) and added calcium complexing agents (phosphate or EDTA). The pH decreased as the temperature was raised from 25 to 90°C and the magnitude of the pH decrease was greater with increase in initial pH at 25°C before heating or milk concentration. The pH decrease on heating from 25 to 90°C in skim milk solutions with added calcium complexing agents was lower than that of milk without the addition of these salts. The calcium activity decreased on heating from 25 to 60°C. The magnitude of the change decreased with increase in initial pH at 25°C before heating and milk concentration. The decrease in calcium activity on heating from 25 to 60°C for skim milk solutions with added calcium complexing agents was lower than that of milk solutions without the addition of calcium complexing agents. The changes in pH and calcium activity on heating milk were largely reversible after cooling the milk. The results suggested that the pH and calcium activity at high temperatures are a function of the milk composition. Knowledge of the initial pH prior to heating alone is not sufficient for predicting the changes that occur during heating.

Nutritional status has a major impact on the immune system. Probiotic effects ascribed to fermented dairy products arise not only from whole microorganisms but also from metabolites (peptides, exopolysaccharides) produced during the fermentation. We recently demonstrated the immunomodulating capacity of kefir in a murine model. We now aimed at studying the immunomodulating capacity in vivo of the products derived from milk fermentation by kefir microflora (PMFKM) on the gut. BALB/c mice received the PMFKM for 2, 5 or 7 consecutive days. IgA+ and IgG+ cells were determined on histological slices of the small and large intestine. IL-4, IL-6, IL-10, IL-12, IFNγ and TNFα were determined in the gut, intestinal fluid and blood serum. IL-6 was also determined in the supernatant of a primary culture of small intestine epithelial cells challenged with PMFKM. PMFKM up-regulated IL-6 secretion, necessary for B-cell terminal differentiation to IgA secreting cells in the gut lamina propria. There was an increase in the number of IgA+ cells in the small and large intestine. The increase in the number of IgA+ cells was accompanied by an increase in the number of IL-4+, IL-10+ and IL-6+ cells in the small intestine. Effects of PMFKM in the large intestine were less widely apparent than the ones observed at the small intestine lamina propria. All cytokines that increased in the small intestine lamina propria, also did so in blood serum, reflecting here the immunostimulation achieved in the gut mucosa. We observed that the PMFKM induced a mucosal response and it was able to up and down regulate it for protective immunity, maintaining the intestinal homeostasis, enhancing the IgA production at both the small and large intestine level. The opportunity exists then to manipulate the constituents of the lumen of the intestine through dietary means, thereby enhancing the health status of the host.

The effect of yogurt on the inhibition of colon tumours induced by 1,2-dimethylhydrazine in BALB/c mice has been studied, and the hypothesis examined that yogurt induces a great reduction in the inflammatory immune response and inhibits tumour growth. Mice were assigned to five experimental groups: a control group fed with a conventional balanced diet, and four other test groups that received yogurt supplements for 2, 5, 7 or 10 consecutive days. At the end of each feeding period, mice were given subcutaneous injections of 1,2-dimethylhydrazine (20 mg/kg) once a week for 8 weeks. After tumour induction, yogurt was given again for 2, 5, 7 or 10 consecutive days each 10 d for 20 weeks. By week 20, 70% of the animals in the control group had developed colorectal tumours. From week 8, there was a considerable infiltration of mononuclear cells into the lamina propria of the large intestine. There was an increase in the number of IgG-producing cells and a slight increase in the IgA-secreting cells, and of CD8+ but not CD4+ T lymphocytes, a high level of β-glucuronidase activity in the intestinal fluid and leucocytosis with neutrophilia in the blood. However, in the test groups given yogurt tumour growth was inhibited, the effect being more evident with 7 or 10 d treatment. The inflammatory immune response as measured by the characteristics we assessed was also reduced, with an increase in the IgA-secreting cells and in CD4+ T lymphocytes. The blood count was similar to that of normal animals and no colorectal tumours were observed in week 20. We suggest that one of the mechanisms by which yogurt exerts antitumour activity is through its immunomodulator activity, by reducing the inflammatory immune response, which was markedly increased when the carcinogen was administered.

The effect of Ca2+. binding and ionic strength on the physicochemical characteristics of phosphoserine residues was studied on O-phospho-DL-serine, bovine β-casein and its phosphopeptide (1–25) using 31P NMR. pK in various experimental conditions were determined.

The pK of the phosphoserine residues of β-casein and its phosphopeptide (1–25) respectively ranged from 6·46 to 7·21 and from 6·57 to 7·10. pK of O-phospho-DL-serine, β-casein and its phosphopeptide decreased when Ca2+ was bound to the phosphoserine residue or when ionic strength was increased. The binding of Ca2+ to the phosphopeptide (1–25) took place at first on phosphoserine residues 17, 18, 19 which had the highest pK, then, when these were saturated, on residue 15 whose pK was the lowest. When the four sites had bound Ca2+, peaks corresponding to a different complex form appeared in the spectrum.

The crystal habit of milk fat is determined by the cooling procedure employed and is changed markedly by interesterification (randomization) of the fat. X-ray diffraction analysis indicated that slowly cooled milk fat occurs in both beta prime and beta modifications, whereas in rapidly cooled milk fat, evidence for the beta prime modification only was obtained. The long spacing was greatly increased by interesterification, especially when the fat was slowly cooled. The presence or absence of certain high melting glyceride fractions greatly influenced solid fat content, softening point and hardness of milk fat.

All 47 non-nisin producing strains of Streptococcus lactis and 12/13 strains of Str. lactis subsp. diacetylactis examined produced bacteriocins, for which the term lactostrepcins is suggested. Seven strains of Str. cremoris examined produced no bacteriocins active against 3 lactic streptococci strains used as indicators. The strains examined were divided into 3 groups: I, those producing lactostrepcins active against only one streptomycin resistant mutant of Str. lactis 60 indicator strain; II, those producing lactostrepcins active against all 3 indicator strains; III, those not producing lactostrepcins active against the indicator strains employed. The lactostrepcins were sensitive to various proteolytic enzymes and to phospholipase D, but retained full or partial activity after dialysis. Most of the bacteriocins studied were fully active only within the pH range 4·2–5·0 and were reversibly inactivated at pH 7·0 or 8·0. Results suggested occurrence of 4 different lactostrepcins. The lactostrepcins produced by all group I strains were the same, but there were differences among the lactostrepcins produced by group II strains. Lactostrepcins killed some β-haemolytic streptococci and some strains of Lactobacillus helveticus. One of the lactostrepcins was also active against certain Leuconostoc strains, but not against other Leuconostoc strains, nor against L. helveticus or other Gram-positive bacteria.

Colostrum and milk feeding are key factors for the newborn ruminant survival, affecting the future performance of the animal. Nowadays, there is an increasing interest in the potential of feeding newborn ruminants (mainly goat kids and lambs) with colostrum and milk from other more productive ruminant species (mainly cows). Although some studies regarding differences between colostrum and milk from these three species have been performed, herein we conduct for the first time a comparison using a proteomics 2-Dimensional Electrophoresis gel-based approach between these three ruminant species. In this study colostrum and milk samples from six Holstein cows, six Canarian sheep and six Majorera goats were used to determine the chemical composition, immunoglobulin G (IgG) and M (IgM) concentrations and proteomics profiles. Results showed that in general sheep colostrum and milk contained higher fat, protein and lactose percentages compared to bovine and goat samples. Additionally, no differences in the IgG or IgM concentrations were found among any of the three studied species, with the exception of sheep colostrum that showed the highest IgM concentration. With reference to the proteomics-based approach, some high abundant proteins such as serum albumin precursor, beta-caseins or different immunoglobulins components were found in colostrum, milk or even both. Nevertheless, differences in other proteins with immune function such as serotransferrin or lactoperoxidase were detected. This study shows that despite the similar immunoglobulin concentrations in colostrum and milk from the three studied species, differences in several immune components can be detected when these samples are studied using a proteomics approach. Finally, this study also provides a base for future investigation in colostrum and milk proteomics and metabolomics.

An investigation was made of the death-rate of Bacillus subtilis spores in suspension in hydrogen peroxide concentrations of 10, 15 and 20% at 25, 50, 60, 70 and 80°C. Average logarithmic survival curves were plotted against normal time and death-rate constants and Q10 values were calculated from the graphically determined decimal reduction times. This treatment of the experimental findings was found to be warranted in the temperature range 60–80°C. The first-order death-rate constant was ∽ 0·1 s−1 for 10% hydrogen peroxide solution at 60°C and the Q10 was about 1·6. Increases in the concentration from 10 to 15% and from 15 to 20% each gave an increase of about 50% in the rate constant. The application of these data to commercial practice is discussed.

Calcium and phosphate concentrations in milk ultrafiltrate prepared at 37 °C over the pH range 4.0–8.0 have been measured and used to calculate the apparent solubility products for various calcium phosphates in milk. Micellar calcium phosphate had a constant Ca/Pi = 1·58·10·03 at all pH values and a near-invariant ion activity product for CaHP04. Apparent solubility products have also been obtained for brushite, hydroxyapatite and amorphous tricalcium phosphate in milk salt solution over a similar pH range. No evidence was found for a hydroxyapatite or amorphous tricalcium phosphate phase in milk.

Experimental cheeses were prepared in triplicate from pasteurized milk inoculated with Debaryomyces hansenii under aseptic conditions. Three cheesemaking replicates, with efficient control of environmental parameters (temperature, relative humidity, atmospheric composition) showed similar ripening characteristics. Deb. hansenii grew only on the cheese surface, where its oxygen demand was satisfied, especially during the first 24 h (mean generation time, 5·8 h). Salting in a sterile saturated brine solution reduced its growth and decreased viability. Growth was slower after 48 h because of the decrease in ripening temperature (mean generation time, 94 h). The total count of Deb. hansenii was maximum (≈3×107 yeasts/mm2) after 6 d ripening and its viable cell concentration was ≈2×106cfu/mm2. This difference was due to the ‘non-viability’ of part of the population. The viable Deb. hansenii concentration was highly correlated (r2>0·95) with the lactate concentration in the inner part and with the surface and inner lactose concentrations, up to day 10 of ripening. This emphasized the importance of the diffusion of carbon substrate from the inner part to the surface of the cheese during ripening. The pH of the inner part depended significantly on the lactate and lactose concentrations. Surface pH was significantly related to inner lactate concentration, temperature and relative humidity. This also demonstrated the controlling role of carbon source diffusion.

Pancreatic lipase deacylation was used to study the variation in distribution of the fatty acids at position sn-2 of milk fat triglycerides of a series of butters and of the milk obtained from a cow on a restricted feeding regimen. Selected samples were analysed for stereospecific distribution of their triglyceride fatty acids which were found to be esterified in a non-random manner. The percentage of a fatty acid at position sn-3 generally decreased with increase in the chain length of the acid; the reverse occurred at position sn-1. There was no major variation in the stereospecific distribution of fatty acids throughout the year; the variation that did occur was exhibited mainly by the medium chain length acids. Variation was greater at positions sn-1 and sn-3 than at position sn-2 and when the proportion of an acid varied at position sn-1 it usually varied at position sn-3 in the reverse manner.

For the milk samples it was found that change in triglyceride fatty acid composition was not associated with random increases or decreases of fatty acids at the 3 stereospecific positions. With changing triglyceride fatty acid composition (6:0, 8:0, 10:0, 12:0, 14:0, and 16:0 decreasing and 18:1 increasing) the percentage of 6:0, 8:0, 10:0, 12:0, 14:0 and 16:0 acids at position sn-3 decreased while the percentage of 18:0 and 18:1 acids increased. These changes were compensated for by changes in the opposite direction at positions sn-1 and sn-2.

It is generally accepted that the udder possesses two main defence mechanisms against mastitis infections. The teat sphincter acts as a barrier against the entry of the pathogenic frequently present on the skin of the teats, and internal factors prevent the bacteria which gain access from setting up an infection. In recent years studies have been made on the relative importance of these two mechanisms, and particular attention has been paid to the relation of teat patency or ease of milking with the incidence of disease. Little (1) stressed the importance of the intact sphincter as a barrier, and suggested that the sphincters of older cows relax, making them more susceptible to infection. In an attempt to overcome the disadvantages of the manual assessment of teat patency, Espe & Cannon (2) used a special instrument. The results were not wholly satisfactory and no correlation with mastitis was found. More recently, Murphy (3) manually examined the teats of at least eighty-seven cows and found a trend towards a higher incidence of infection in quarters with patent teats. This latter author did not agree that relaxing of the teat sphincters with advance in age could account for the increased susceptibility of older cows to streptococcal mastitis (4). While Murnane (5) could not confirm Murphy's results, McEwan & Cooper (6), again using the manual method, obtained further evidence definitely suggesting that easy milking cows were more prone to mastitis. Although Stuart & Lancaster (7) found evidence confirming the importance of the teat orifice, they concluded that it was of less importance than those factors operating within the udder.