Six batches of Casar de Cáceres raw ewes' milk cheeses were examined throughout their ripening. Three were made in winter, the others in spring. The microbial counts in the interior of the cheeses were ∼1·5 log units higher in the winter cheeses for total viable counts, lactic streptococci and leuconostocs, and >2 log units higher for total enterobacteria, coliforms and faecal coliforms in the 60 d samples. Staphylococci were not detected after day 15 of maturation in spring cheeses, but reached 1 log unit at day 60 in the interior of winter cheeses. In the surface samples, differences in microbial counts were more marked. Values of some physicochemical indices, such as pH, % moisture content and % NaCl were also determined.

Constituents of a flavour concentrate from fishy butterfat were separated by gas chromatography on silicone oil and ‘Carbowax 400’ columns. Two fractions with distinct flavours were obtained, the one of oily flavour and containing n-hexanal, n-heptanal, hex-2-enal and heptan-2-one, the other of metallic flavour, containing a single carbonyl compound present in relatively small amount. It is considered that these compounds with the exception of heptan-2-one are mainly responsible for the characteristic fish-oil flavour.

Other compounds isolated include propanal, n-pentanal, n-octanal, n-nonanal, n-decanal, acetone, pentan-2-one, nonan-2-one, undecan-2-one, acraldehyde, pent-2-enal, hept-2-enal, oct-2-enal, non-2-enal, hepta-2, 4-dienal, a compound resembling octa-2, 4-dienal, a compound with a mushroom flavour, and a compound that formed the 2, 4-dinitrophenylosazone of methylglyoxal.

This study investigated the application of near infrared (NIR) reflectance spectroscopy to the measurement of texture (sensory and instrumental) in experimental processed cheese samples. Spectra (750 to 2498 nm) of cheeses were recorded after 2 and 4 weeks storage at 4 °C. Trained assessors evaluated 9 sensory properties, a texture profile analyser (TPA) was used to record 5 instrumental parameters and cheese ‘meltability’ was measured by computer vision. Predictive models for sensory and instrumental texture parameters were developed using partial least squares regression on raw or pre-treated spectral data. Sensory attributes and instrumental texture measurements were modelled with sufficient accuracy to recommend the use of NIR reflectance spectroscopy for routine quality assessment of processed cheese.

The 31P nuclear magnetic resonance spectrum of liquid milk was examined. Of the three peaks observed, the two larger were assigned to inorganic phosphate (Pi) and the seryl phosphate (SerP) residues of casein; the third peak was assigned to a phosphodiester, which is probably glycerophosphoryl choline. The pH-dependences of the chemical shifts of the Pi and SerP were measured with and without added EDTA, and the results confirm the assignments. The width of the Pi peak in milk is significantly greater than in similar solutions lacking casein, probably because of binding to, or chemical exchange with, the casein micelle. Most of the SerP residues in milk are not sufficiently mobile to have been detected in these experiments but a significant fraction of SerP residues is able to move freely and can be titrated.

Indirectly heated ultra-high-temperature (UHT) processed milk was prepared with initially high, medium, and low dissolved O2 contents of 8·9, 3·6 and 1·0 ppm respectively, aseptically bottled, and tested at intervals during storage at room temperature for 150 d. Flavour acceptability increased to a maximum after a few days, but declined slowly after about 6 d; the increase was associated with less off-flavour described as ‘cabbagey’, and the decrease with more ‘stale’ off-flavour descriptions. Milks with higher initial O2 contents were preferred up to 8–13 d, but thereafter acceptability was independent of initial O2 content. Sulphydryl group (–SH) contents rapidly decreased and O2 levels correspondingly declined in the first few days as the flavour improved. Loss of –SH was lower with lower initial O2 contents, and moderate –SH content remained in low O2 samples for several weeks. Ferricyanide reducing (FR) values did not satisfactorily measure stale flavour development. They were initially high and decreased during the first 13 d at rates dependent on O2 content. After 20 d the FR values began to rise in high O2 samples, but continued to decline slowly in low O2 samples up to 90d although stale flavour was increasing.

High initial O2 contents resulted in rapid depletion of ascorbic acid and folic acid during storage. Losses of vitamin B12 were small, but were higher with high O2 contents than with low.

The beneficial effect of O2 on flavour, therefore, appears to be so slight and confined to such a short period in the early life of the milk as to be completely outweighed by the adverse nutritional effects.

Starting in week 5 of lactation, goats were milked unilaterally once daily for 4 ½ weeks, then twice daily for 12 weeks and finally thrice daily for 2 weeks. The control gland was milked twice daily throughout. During less frequent milking, yield fell by 26%. There was a rapid and essentially complete recovery of milk yield on resumption of twice-daily milking, and five of the six goats subsequently responded to thrice-daily milking with a small increase in milk yield. Mammary function was assessed during each phase by determining enzyme activities and in vitro synthesis rates of lactose and casein in mammary biopsies. There was a trend towards reduced differentiation of secretory cells during infrequent milking, with very little recovery during twice-daily milking but complete recovery during thrice-daily milking. The differentiative state of the control gland remained unchanged throughout.

Preparation, physicochemical characterization and stability of iron-supplemented caseins were studied. When ferrous iron (final concentrations of 0·25, 0·5, 1·0 and 1·5 mM) was added to sodium caseinate (final casein concentration 25 g/l), iron binding to caseins was complete. In parallel, an increase in absorbance at 280 nm, a decrease in fluorescence intensity, modifications of reversed-phase HPLC profile, a decrease in pH and an increase in free sodium concentration were observed. Moreover, no release of iron from iron-casein complexes was found between pH 6·5 and 3·7 or after heat treatments or in the presence of inorganic phosphate. Only EDTA disodium salt and trisodium citrate had significant effects on the release of iron bound to caseins.

Colostrum and milk samples from 20 goats were analysed for concentrations of immunoglobulin G (IgG), β-lactoglobulin (β-lg), α-lactalbumin (α-la) and serum albumin (CSA) throughout the first 14 milkings post partum (7 d of lactation) using single radial immunodiffusion assay. Concentrations (mg/ml, means±SD) at first milking were IgG 47·9±25·5, β-lg 30·7±10·4, α-la 2·77±0·82 and CSA 2·97±2·46 mg/ml. Large variations were recorded for IgG concentrations (19·9–94·5 mg/ml) and β-lg (9·3–49·8 mg/ml). Concentrations of IgG, β-lg and CSA dropped abruptly in the subsequent milkings and α-la concentration decreased slowly. Mean IgG concentration was <2 mg/ml after 7 milkings and <1 mg/ml after 11 milkings. However, IgG concentration does not differ significantly, at the 1% level, from milkings 7–14. The contribution of β-lg to the increase in whey proteins in early milks was greater than that of IgG from milkings 5 to 14. The results were tabulated to make it possible to calculate the excess of whey proteins that would be obtained if early milks were illegally added to milk supply.

An analytical procedure, based upon HPLC, has been used to determine the degree of isomerization of retinol (vitamin A) in various dairy products. In raw milks, which are not subjected to thermal processing, there was no conversion of the predominant all-trans-isomers to cis-isomers in samples from various species. Pasteurized milks with mild heat treatment (high quality milk) had an average 13-cis[ratio ]all-trans ratio of 2·6%, while pasteurized milk treated for 15 s at temperatures ranging from 72 to 76°C had an average ratio of 6·4%. Milk subjected to more severe heat treatments had a higher degree of isomerization (UHT milk, 15·7%; sterilized milk, 33·5%), consistent with increased thermal conversion of the retinol isomers. In pasteurized and UHT cream, the increase in 13-cis-isomer was also a consequence of heat treatment (pasteurized cream, 3·0%; UHT cream, 14·4%). The presence of cis isomers in fermented milk suggests that fermentation processes, directly or indirectly, can induce cis–trans isomerization. In the cheeses analysed, the extent of retinol isomerization ranged from 7·6 to 35·0%. Our results confirm the importance of individual isomers in evaluating the vitamin A status of dairy products.

The addition of simple aldehydes brought about large increases in the heat stability of both skim-milk and concentrated skim-milk over a comparatively wide milk–pH range. The coagulation time–pH minima of type A milks were completely removed by aldehyde treatment. Some sugars, which react readily as aldehydes on heating, were also shown to stabilize concentrated milk to prolonged heat treatment at 120 °C.

Staphylococcus aureus isolates produce several pathogenic factors. The combination of these products influences the pathogenic role of different isolates, but their specific effects are well known in the pathogenesis of udder infections. This study focused on the association of polymorphism of the coagulase gene, protein A gene, collagen-binding protein gene, and of fibrinogen-binding protein gene on somatic cell count (SCC) and on Staph. aureus growth rate. Fifty Staph. aureus isolates from 13 dairy cow herds, located in seven different provinces, were considered. The results showed a low frequency of cna gene, similar to the one observed in human isolates. Meanwhile, the high frequency of efb gene indirectly confirmed the role of this factor in bacterial pathogenesis, being associated with adhesion to epithelia. The association of these two single genes with SCC and growth rate showed to be not significant. The polymorphism of spa gene was confirmed to be significantly associated with inflammatory response and growth rate, albeit with a pattern different from the one suggested for human isolates. Sorting of isolates based on the clusters obtained by combining polymorphisms of spa and coa genes and the presence of cna and efb genes, showed that a single cluster (cluster V) was prevalent in the different herds and provinces, while the other six clusters identified were widely spread among the remaining 60% of the isolates. Results showed that clusters VI and VII had significantly higher growth rates at 3, 4, and 6 h in comparison with the other clusters. Meanwhile, quarters infected with these strains showed significantly lower SCC levels. The frequency of isolates from cluster V, suggested that they should possess pathogenic factors increasing their invasiveness, even if in the presence of a stronger inflammatory response. These results indirectly confirm previous findings on the different interactions between isolates and the udder immune system. They also suggest that isolates with higher growth rates and inducing a lower inflammatory response have better chances to spread among the herd. The relatively simple genomic method proposed in this study could be applied by an increasing number of diagnostic laboratories and could be useful in studying the epidemiology of Staph. aureus intramammary infections in dairy herds when collecting data from the field.

This review provides an overview of the composition, structure, and biological activities of milk fat globule membrane (MFGM) compounds with focus on the future application of this compound as a food ingredient. MFGM is a particular component of mammalian milks and is comprised of a tri-layer of polar lipids, glycolipids and proteins. In recent years, MFGM has been extensively studied for the purpose of enhancing the efficacy of infant nutrition formula. For example, infant formulas supplemented with bovine MFGM have shown promising results with regard to neurodevelopment and defense against infections. Components of MFGM have been shown to present several health benefits as the proteins of the membrane have shown antiviral activity and a reduction in the incidence of diarrhea. Moreover, the presence of sphingomyelin, a phospholipid, implies beneficial effects on human health such as enhanced neuronal development in infants and the protection of neonates from bacterial infections. The development of a lipid that is similar to human milk fat would represent a significant advance for the infant formula industry and would offer high technology formulas for those infants that depend on infant formula. The complexity of the structure of MFGM and its nutritional and technological properties is critically examined in this review with a focus on issues relevant to the dairy industry.

Simultaneous measurement of viscosity and degree of proteolysis of κ-casein, α, in skim milk at three different concentrations of rennet and three different concentrations of casein showed that viscosity was a function of α until the former reached its minimum value. Then the relation between viscosity and α depended on the enzyme concentration. This is considered to be caused by casein micelles starting to aggregate at this point. The minimum in viscosity was found when α was between 0·6 and 0·7, indicating that casein micelles started aggregating at a lower degree of proteolysis than was believed earlier. The rate of gel firming was found to depend on the concentration of added rennet even after κ-casein was completely proteolysed. The results demonstrated that both proteolysis of κ-casein and aggregation of casein micelles must be taken into account when modelling the renneting reaction.

Tryptic phosphopeptides were obtained from whole bovine casein by chromatography on the anion exchange resin QAE-Sephadex A 25. Salt gradient elution of the column allowed separation of non-phosphorylated peptides from phosphorylated species. The preparations obtained contained at least seven distinct phosphopeptides of which the following casein fragments were identified: αs1(43–58):2P, αs1(59–79): 5P, αs2(46–70): 4P, β(1–28): 4P, β(2–28): 4P, and β(33–48): 1P. Fast protein liquid chromatography (FPLC) on Mono Q HR 5/5 resin showed that the phosphopeptides were eluted in the same order as from the QAE-Sephadex resin. However, on the analytical column HR 5/5 the fragments αs1(59–79): 5P and β(2–28): 4P, having the same net charge under the conditions of chromatography, co-eluted, whereas they were at least partly separated on the preparative column HR 16/10. Following enzymic dephosphorylation, the peptides eluted at lower salt strength in the gradient. FPLC on Mono Q resin thus permitted dephosphorylation to be monitored and intermediates between the parent species and the fully dephosphorylated peptide to be identified.

Trans fatty acids (TFA) formed during biohydrogenation by ruminant animals were quantified in cows' milk fat by means of Fourier transform i.r. (FTIR) spectroscopy. When samples were analysed as the derived methyl esters, a spectral subtraction technique resulted in TFA values that were not biased by the unspecific absorption of intact triacylglycerols or influenced by the shape of the trans band. Austrian milk fat samples contained 20–50 g TFA/kg. Results obtained by analysis of TFA as triacylglycerols compared favourably with those obtained by using methyl esters. The findings of the FTIR method were further substantiated by applying argentation chromatography in combination with capillary gas chromatography to the same set of samples. Besides trans monoenoic acids, non-methylene interrupted cis, trans octadecadienoic acids contributed significantly to the total trans content. Two novel TFA, trans-11, cis-15 octadecadienoic and cis-9, trans-13 octadecadienoic acids, were identified in milk fat by means of their retention behaviour on argentation chromatography and gas chromatography as well as by an oxidative degradation procedure.

Several strains of Lactococcus lactis subsp. lactis, Lactobacillus casei and Lactobacillus plantarum isolated from traditional goats' cheese have been studied for titratable acidity, proteolysis in milk and enzymic activities. Aminopeptidasc activities were measured with whole cells and cells permeabilized with Triton X-100. Caseinolytic activity was investigated using electrophoresis in polyacrylamide gel with sodium dodecyl sulphate. Lc. lactis subsp. lactis had a level of proteolytic activity in skim milk greater than that of Lb. casei, while this activity in Lb. plantarum was very low. Alanine aminopeptidase activity was almost non-existent for all strains tested, while lysine aminopeptidase activity appeared to be of fundamentally intracellular origin. Leucine aminopeptidase activity was also greater in cells that had been permeabilized than in whole cells for Lb. casei and Lb. plantarum. Lc. lactis subsp. lactis leucine aminopeptidase activity was greater in whole cells. No significant hydrolysis of casein was found with Lb. casei I FPL 725 and Lb. plantarum IFPL 722 permeabilized with Triton X-100 after 24 h incubation with whole bovine casein.

The present study identifies proline-specific peptidases found in Streptococcus cremoris AM2. These activities were present prcdominantly in the cytoplasmic fraction of the cells with minor amounts in cell membrane fraction. No evidence vvas found for such activities in either the extracellular fluid used to grow the cells, in the wash fraction or in the cell wall fraction. The involvement of these enzymes in release of free proline from proline-containing peptides is considered.

An outstanding parameter in cheese making is the type of coagulant, which greatly influences the characteristics of the final products. Proteolysis is the most important set of biochemical changes during ripening of most cheeses, and is carried out, in different magnitude, by proteolytic agents originated in milk, rennet (or rennet substitute), and starter and non-starter micro-organisms (Silva & Malcata, 2000). The demand for alternative sources of milk coagulants, to replace the expensive and limited natural rennet supplies, has increased (Esteves et al. 2001). All commercial enzymes employed as milk coagulant are aspartic proteinases, which are most active at acidic pH and preferentially cleave peptide bonds between residues with hydrophobic side-chains (Silva & Malcata, 1999).

We investigated the mucus-binding properties of aggregating and non-aggregating potentially probiotic strains of kefir-isolated Lactobacillus kefiri, using different substrates. All the strains were able to adhere to commercial gastric mucin (MUCIN) and extracted mucus from small intestine (SIM) and colon (CM). The extraction of surface proteins from bacteria using LiCl or NaOH significantly reduced the adhesion of three selected strains (CIDCA 8348, CIDCA 83115 and JCM 5818); although a significant proportion (up to 50%) of S-layer proteins were not completely eliminated after treatments. The surface (S-layer) protein extracts from all the strains of Lb. kefiri were capable of binding to MUCIN, SIM or CM, and no differences were observed among them. The addition of their own surface protein extract increased adhesion of CIDCA 8348 and 83115 to MUCIN and SIM, meanwhile no changes in adhesion were observed for JCM 5818. None of the seven sugars tested had the ability to inhibit the adhesion of whole bacteria to the three mucus extracts. Noteworthy, the degree of bacterial adhesion reached in the presence of their own surface protein (S-layer) extract decreased to basal levels in the presence of some sugars, suggesting an interaction between the added sugar and the surface proteins. In conclusion, the ability of these food-isolated bacteria to adhere to gastrointestinal mucus becomes an essential issue regarding the biotechnological potentiality of Lb. kefiri for the food industry.

1. Thirty-five strains of acetoin producing lactic streptococci isolated from raw milk, from starter cultures and from dairy products were investigated biochemically and serologically.

2. The strains are shown to form a homogeneous unit, very similar to Strep. lactis O.J., which species they resemble in cultural behaviour, in growth-temperature range, in tolerance of sodium chloride and heat and in carbohydrate and other fermentations. They differ from Strep. lactis essentially in their ability to utilize citrate in producing CO2 volatile acids and C4-compounds (diacetyl, etc.). With few exceptions they bear no resemblance to the enterococci.

3. Serologically the strains are shown to belong to group N and thus the relationship to Strep. lactis is clearly established.

4. The strains investigated are shown to be identical with Strep. citrophilus van Beynum & Pette and almost with certainty, though no direct comparison has yet been possible, with Strep. diacetilactis Matuszewski et al. Probably they are identical, too, with Strep. diacetyl aromaticus Karnad and Strep. lactis aromaticus Joshi & Ram Ayyar.

5. The taxonomic position of the relevant unit is discussed, and it is suggested that it be given species rank under the name Strep. diacetilactis.