Kefir is a traditional fermented milk originating many centuries ago in the Caucasian mountains. It is produced by fermentative activity of ‘kefir grains’ consisting mainly of lactococci, lactobacilli and yeasts in a protein–polysaccharide matrix. The grains contain a relatively stable and specific balance of microorganisms which exist in a complex symbiotic relationship. The grains grow in the process of kefir making only from pre-existing grains (Saloff-Coste, 1996). When kefir grains are allowed to grow in milk, microorganisms are shed from the grains into milk where they continue to multiply with the production of acid, flavour and physicochemical changes.

The traditional method of kefir making is currently by adding kefir grains directly as starter to milk that has been pasteurized and cooled to 20–25°C. After a period of fermentation lasting ∼24 h, the grains are removed by filtration and the beverage is ready for consumption (Saloff-Coste, 1996). Kefir from which the grains have been removed may be used as starter. However, this fermented milk cannot be used for subsequent inoculations to make an acceptable product, because the original balance of microorganisms has been disrupted (Kroger, 1993).

The complex microbiological composition of kefir grains explains why it is difficult to obtain starter with the optimal and constant composition necessary for a regular production of kefir of standard quality (Koroleva, 1988a). Studies have been undertaken to establish cultivation conditions[ratio ]grain[ratio ]milk ratio, cultivation temperature, period of time and conditions prior to separation of grains from the fermented milk, shaking conditions for agitation of milk with the grains in the course of fermentation, washing of kefir grains and so on. All these factors influence the microflora of the kefir starter and fermented milk. There are no rules about household manufacture of kefir. Different reports indicate a wide range of grain[ratio ]milk ratios for kefir making. Bottazzi & Bianchi (1980), Marshall & Cole (1985), Merin & Rosenthal (1986), Mann (1989), Hosono et al. (1990) and Kroger (1993) employed 20–50 g/l while Koroleva (1988a) employed 20–100 g kefir grain/l and Marshall et al. (1984) and Neve (1992) 50–100 g/l. Rea et al. (1996) used 1 g kefir grain/l as starter and 200 g starter in the form of kefir grains is recommended by Hansen for the fermentation of 1 l milk (Marshall & Cole, 1985). A critical control point in kefir manufacture to obtain a product with constant quality is the standardization of the kefir grain[ratio ]milk ratio. Koroleva (1988b) claimed that it is better to use kefir grains as starter for kefir production and, at the same time, to decrease the amount of inoculum.

The purpose of this study was to evaluate the effects of changes in the kefir grain[ratio ]milk ratio (quantity of kefir grains inoculated into the milk) on microflora composition, acidity, apparent viscosity and carbon dioxide content of fermented milk.

Fouling from milk fluids is a severe industrial problem which reduces the efficiency of process plant. The chemistry of fouling has been thoroughly investigated but the sequence of events that occur is not yet clear. Deposit contains both protein and minerals. Experiments have been carried out to determine the sequence of events in the fouling of stainless steel surfaces at 96 °C from turbulent flows of whey. Contact times between 4 and 210 s have been studied, and surface analysis techniques used to detect the distribution of elements. The first layer of deposit, formed after 4 s of contact between the fluid and the surface (fluid temperature 68 and 73 °C), consisted mainly of protein and was identified by X-ray photoelectron spectroscopy analysis. There was a lag phase of up to 150 s for a fluid temperature of 73 °C before deposit aggregates were observed to adsorb on to the surface. These aggregates were identified as protein and Ca by X-ray elemental mapping. No P was found in any experiments for this exposure. After 60 min contact time, however, both Ca and P were found at the interface between deposit and the stainless steel surface, irrespective of the Ca and P content of the test fluid.

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.

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.

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.

The trypsin-inhibitor capacity of bovine milk was seen to increase in mastitits. This capacity showed good correlation with the California Mastitis Test score, somatic cell count (Coulter Counter method) and bovine serum albumin (BSA) content (radial immunodiffusion). The elevated antitrypsin level proved to be a safer indicator of mastitis than BSA alone. Using gel-filtration chromatography, the major trypsin-inhibitor activity of mastitic milk and plasma was eluted with BSA, indicating a similar mol. wt (70000). This fraction showed electrophoretic polymorphism. A large molecular weight fraction cross-reacted immunologically with human α2-macroglobulin. The antitrypsin activity in mastitic milk was clearly different from the principal activity in colostrum, which had a mol. wt of about 13000.

Twelve Lactobacillus isolates from Brazilian starter-free ovine cheeses were evaluated for their probiotic potential. The strains were identified by 16S rDNA sequencing as Lactobacillus plantarum (7), Lb. brevis (2), Lb. casei (2) and Lb. parabuchneri (1). All strains showed variable resistance to gastric juices and relative tolerance to pancreatin and bile salts. Only five strains of Lb. plantarum could not deconjugate the sodium salt of taurodeoxycholic acid. Autoaggregation ability after 24 h was above 50% and hydrophobicity was higher than 60% for most strains. All lactobacilli could inhibit linolenic acid oxidation, except Lb. parabuchneri strain, whereas none of them could scavenge DPPH radical. β-Galactosidase activity ranged from 47·7 to 2503 Miller units. Inhibition of food pathogens Listeria monocytogenes, Staphylococcus aureus, Bacillus cereus, Escherichia coli and Salmonella typhimurium was demonstrated and the production of organic acids could be associated with this effect. The Lactobacillus strains from Brazilian regional ovine cheese showed interesting functional characteristics, mainly the strains Lb. brevis SM-B and Lb. plantarum SM-I. Both presented high acid tolerance. In addition, Lb. brevis SM-B also displayed remarkable antioxidant activity and Lb. plantarum SM-I was the highest β-galactosidase producer, exhibited high autoaggregation and hydrophobicity properties.

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.

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.

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.

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.

Blood samples, taken on 6 occasions during 1 year from 172 cows resident in a dairy herd, were analysed for packed cell volume, blood glucose, haemoglobin (Hb), serum albumin, urea N, total protein, Ca, inorganic phosphate, Mg, Na and K. The data were analysed statistically, firstly with respect to date of sampling, stage of pregnancy or lactation and the interaction between stage of pregnancy or lactation and date of sampling, and secondly with milk yield as an additional factor for the data for lactating cows.

Concentrations of all constituents except inorganic phosphate and K varied significantly (P<0·001) with stage of lactation and/or pregnancy. The most significant changes in concentrations were confined to the periods up to 3 months either side of calving. The greatest changes in concentration during these periods occurred for Mg which rose during late pregnancy and for albumin which fell at or near calving. Albumin, urea and glucose were all, on average, lowest in concentration during the first month of lactation; globulin concentrations showed the reverse trend. Haemoglobin concentrations decreased during late pregnancy and early lactation and were lowest in the period 30–120 d post partum. Calcium concentrations decreased during late pregnancy and rose during early lactation. Sodium concentrations rose during late pregnancy. Interactions between dates of sampling and stages of lactation or pregnancy were significant (P<0·001) for all constituents, and most marked for glucose, urea, albumin, Hb, Ca and Na.

Haemoglobin and K concentrations fell and Mg concentrations rose with increasing milk yield, but the proportions of the total variance accounted for by differences in milk yield were small in relation to those accounted for by stage of lactation. These findings are of significance in relation to the selection of animals for sampling in the Compton Metabolic Profile Test.

Five ovine αsl-casein variants (A–E) were identified in an Italian population sample using gel electrophoresis at alkaline pH, gel isoelectric focusing, two dimensional gel electrophoresis, and immunoblotting with polyclonal antibodies against αs1-casein. Each casein sample produced two peaks by fast reversed-phase HPLC. Gel isoelectric focusing and electrospray mass spectrometry were used to demonstrate that the first HPLC peak contained the 191 residue αsl-casein molecular species and the second the 199 residue species, in proportions of ∼20:80. Only in the case of the sample containing αsl-casein CE was the method for the separation of the single short and long forms of each variant unsuccessful. Both two dimensional electrophoresis followed by staining with polyclonal antibodies against αsl-casein and electrospray mass spectrometry showed a heterogeneity consistent with that expected from a protein chain with three levels of phosphorylation and two different lengths. However, especially for αsl-caseins D and E, a further uncharacterized heterogeneity was detected.

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.

Whole casein preparations obtained from paired quarters (one healthy and one subclinically infected) of each of 6 individual cows were fractionated quantitatively by column chromatography on hydroxyapatite. Infection was associated with an apparent decrease in the αs- and β-casein fractions and increase in the γ- and κ-casein fractions. The increase in the κ-casein fraction was attributable mainly to enrichment with chymosin-resistant minor components of casein.

The profile produced by column chromatography on DEAE-cellulose of casein isolated from a healthy quarter of a single cow differed significantly from that from a subclinically infected quarter of the same cow. These differences were increased by incubation of the quarter milks at 37 °C for 8 h. Extensive fragmentation of [14C]methylated αs2- and β-casein, and less extensive fragmentation of [14C]methylated αs1-casein, occurred during incubation of these proteins in milk from an infected quarter. Fragmentation was almost entirely inhibited by soybean trypsin inhibitor. From a consideration of the chromatographic behaviour of the radiolabeled fragments on DEAE-cellulose, and other data presented, it was concluded that proteolysis of caseins by a plasmin-like enzyme was sufficient to account, in qualitative terms, for most of the compositional changes exhibited by casein from subclinically infected quarters.

Olive tree leaves (OTL) and grape marc (GM) are by-products with high linolenic (LNA) and linoleic (LA) acid content, respectively, which can be used as dietary ingredients to increase the cis-9 trans-11 conjugated linoleic acid (CLA) content of milk fat in sheep and goats. An experiment was conducted with 16 Friesian ewes and 16 Alpine goats to study the effect of OTL and GM inclusion in sheep and goat diets on their milk fatty acid profile, with emphasis on cis-9, trans-11 CLA and vaccenic acid (VA). Ewes and the goats were fed the control (C) diets from parturition to the 90 days in milk (DIM) and then both groups were divided into two sub-groups (treatments). The control groups of both species continued to be fed the C diets, whereas the treated groups were gradually switched over a 2-week period (DIM=91–105) from the C diets to that of treatment 1, which contained air-dried OTL. These OTL diets were fed ad libitum for 1 month (DIM=106–135). After that period, the same treated groups, after 2 weeks of gradual adaptation (DIM=136–150), were switched to treatment 2, which contained air-dried GM. The GM diets were fed ad libitum for 1 month (151–180 DIM). Concentrations of polyunsaturated fatty acids (PUFA) and mono-unsaturated fatty acids increased significantly in milk fat of sheep fed OTL v. C. For goats, only the PUFA in milk fat was increased by feeding OTL compared with C. Relative to C, GM increased significantly the concentration of PUFA only in milk fat of sheep. OTL and GM diets increased the cis-9, trans-11 CLA and VA content in milk fat, compared with C, only in sheep. GM caused a sharp increase in 18:0 only in sheep milk fat, while the OTL diet increased significantly the 18:0 in milk fat of goats. GM and OTL diets also had opposite effects on the 18:1/18:0 ratio of sheep milk fat. In conclusion, OTL and GM, when included in sheep diets altered the milk fatty acid profile with a pronounced increase in cis-9, trans-11 CLA and VA contents. The results show that the response of sheep and goats to OTL and GM diets was different, suggesting a species difference that needs further investigation.

In five short-term experiments conducted in Victoria in 1997 and 1998, grazing dairy cows were given either pasture alone or pasture supplemented with high-energy concentrates, and the fatty acid profiles of milk fat were measured. We established the effects of these feeds on some aspects of milk fat of importance for human nutrition, but we specifically focused on the hypothesis that conjugated linoleic acid (CLA) concentrations in milk fat increase as pasture intake increases, and decrease as more concentrates are fed. In agreement with previous research, feeding fresh pasture alone resulted in high concentrations (1·0–1·8 g/100 g milk fat) of CLA. When the effect of level of pasture consumption on CLA content was examined, a significant positive relationship (r2=0·35; P<0·05) was obtained. When cereal grain concentrates were used to supplement pasture intake, the CLA content of milk fat generally declined (P<0·05), except where the amount of concentrates given led to a marked reduction in total milk fat concentration. The use of cereal grain concentrates also generally resulted in significant (P<0·05) increases in medium-chain saturated fatty acids, but always reduced the contribution of butyric acid to milk fat, from 4·5 to 3·9 g/100 g milk fat, on average.

For its characteristics, donkey milk has been proposed as an alternative to goat or artificial milk to feed allergic infants. Therefore, it is important to increase our knowledge on health and immunological characteristics of donkey milk. Ten donkeys, bred as companion animals, were enrolled in this study and sampled once a month, for eight months. Milk (10 ml) was collected from each half udder for somatic cell count (SCC), bacteriological analysis and total bacteria count (TBC). The major pathogens were tested for antimicrobial susceptibility, and Staphylococcus aureus isolates were further genotyped by nanoarray analysis. Whey lysozyme and NAGase (NAG) activities were also assessed. Overall, 101 half-udder milk samples were taken. They showed very low values of TBC (<250 cfu/ml) and SCC (<50 000 cells/ml) and a minor prevalence of pathogens: Staph. aureus was isolated only from 5 milk samples (3 animals), Streptococcus equi from 2 samples and Str. equisimilis from a single sample. All the isolates were sensitive to all antibiotic classes used in veterinary medicine. None of the Staph. aureus isolates were shown to harbour genes coding for any enterotoxin, toxic-shock syndrome toxin or antibiotic resistance. Lysozyme levels were always very high (4000–5000 U/ml), while NAG values were mostly low (<50 U/ml), out of the last part of lactation. The results of this study confirmed the low prevalence of intramammary infections in donkey and the absence of food-borne pathogens, suggesting that donkey milk could be a safe food, if the mammary gland is healthy and the animals are milked in proper hygienic conditions.