The intestinal ecosystem contains a normal microbiota, non-immune cells and immune cells associated with the intestinal mucosa. The mechanisms involved in the modulation of the gut immune system by probiotics are not yet completely understood. The present work studies the effect of a fermented milk containing probiotic bacterium Lactobacillus (Lb.) casei DN114001 on different parameters of the gut immune system involved with the nonspecific, innate and adaptive response. BALB/c mice received the probiotic bacterium Lb. casei DN114001 or the probiotic fermented milk (PFM). The interaction of the probiotic bacteria with the intestine was studied by electron and fluorescence microscopy. The immunological parameters were studied in the intestinal tissue and in the supernatant of intestinal cells (IC). Results showed that the probiotic bacterium interact with the IC. The whole bacterium or its fragments make contact with the gut associated immune cells. The PFM stimulated the IC with IL-6 release, as well as cells related to the nonspecific barrier and with the immune cells associated with the gut. This last activity was observed through the increase in the population of different immune cells: T lymphocytes and IgA+ B lymphocytes, and by the expression of cell markers related to both innate and adaptive response (macrophages). PFM was also able to activate the enzyme calcineurine responsible for the activation of the transcriptional factor NFAT. PFM induced mucosal immune stimulation reinforcing the non-specific barrier and modulating the innate immune response in the gut, maintaining the intestinal homeostasis.

The distribution and activity of alkaline phosphatase (E.C. 3.1.3.1), acid phosphatase (E.C. 3.1.3.2), catalase (E.C. 1.11.1.6), xanthine oxidase (E.C. 1.2.3.2), aldolase (E.C. 4.1.2.7 and 4.1.2.13), ribonuclease (E.C. 2.7.7.16) and carbonic anhydrase (E.C. 4.2.1.1) were studied in the major components of bovine milk. Fractionation was accomplished by centrifugation of milk, skim-milk and buttermilk, and ammonium sulphate precipitation of skim-milk serum. The range of activities found for the enzymes studied are tabulated together with the activities of some of the enzymes in mastitic milk, and the significance of the results obtained is discussed. No carbonic anhydrase activity was detected in any of the samples tested. The other enzymes studied were found to have a greater proportion of their total activity located in the skim-milk fraction. However, all of these enzymes except ribonuclease had a higher specific activity in the fat fraction.

Lactoperoxidase activity and thiocyanate content were monitored in the milks of seven individual cows and six goats during lactation. Lactoperoxidase activity exhibited a cyclic pattern with alternating peaks and troughs throughout lactation. Extremely large variations were observed between and within animals. For example, lactoperoxidase activity ranged from 0·05–5·60 U/ml for one cow on different sampling days. Variations between cows were also large, ranging from 0·98–5·1 U/ml on a specific sampling day. Mean lactoperoxidase activity in cows ranged from 1·5–2·7 U/ml with an overall mean of 2·3±1·0 U/ml while the thiocyanate concentration ranged from 6·0–10·2 μg/ml with an overall average of 8·5±5·1 μg/ml. Lactoperoxidase activity means for goats ranged from 0·04–0·16 U/ml with a grand mean of 0·1±0·06 U/ml while the thiocyanate content means ranged from 6·6–8·2 μg/ml with a grand mean of 7·0±2·59 μg/ml. There was no correlation between lactoperoxidase activity and thiocyanate content in either cows' (R2 = 0·011) or goats' (R2 = 0·015) milk. These experiments have revealed that lactoperoxidase activity is affected by many factors including the individual animals, species, feed and stage of lactation. Therefore the exogenous supply of thiocyanate and hydrogen peroxide needed to activate the lactoperoxidase system for raw milk preservation will vary in quantity depending on these factors.

The effect of physicochemical characteristics (pH, temperature, composition) of the aqueous phase on the mineral balance in milk and milk retentate has been studied. The ratio of colloidal Ca to total protein decreased with pH, but at any given pH the higher the protein concentration, the higher was the ratio of colloidal Ca to total protein. The solubilization of Ca during cooling and the decrease in soluble Ca during heating were approximately the same in retentates and in milk. Among the components of the aqueous phase, soluble Ca and citrate ions were related to the amount of colloidal Ca.

1. A statistical analysis has been made of lactation records from 500 dam-daughter pairs of the Ayrshire breed. Measurements were taken of milk yield, fat, S.N.F., crude protein, casein and lactose.

2. A correction was made for the age of the cow. This contributed a substantial fraction of the total variation for milk yield, S.N.F. and lactose contents. Month of calving had a trivial effect on the lactation average for the characters when compared with the individual variation between cows.

3. After correction for age, the variation between lactations was highest for fat content (standard deviation 0·35%) and slightly less for S.N.F. (0·26%).

4. The heritability was estimated by the dam-daughter correlation within herds. Values in the neighbourhood of 0·5 were obtained for S.N.F., crude protein, casein and ‘ash’ (determined by difference), about 0·35 for fat and lactose, 0·25 for milk yield and 0·13 for non-casein nitrogen.

5. The phenotypic correlations between yield and lactose were positive for both dam and daughter groups. The correlations between yield and the other constituents were more negative in the daughter group than in the dam group. The correlation between yield and S.N.F. was, in fact, positive in the latter. The correlation between fat and S.N.F. averaged 0·36, being derived mainly from the correlation between fat and casein (0·43), though that with lactose was also positive (0·13).

6. The variation in S.N.F. was mostly determined by that in casein (r=0·85). There was a positive correlation between lactose and casein (0·24), and a negative one between lactose and non-casein nitrogen (–0·41). Lactose was negatively correlated with ‘ash’ (–0·31). ‘Ash’ was positively correlated with non-casein nitrogen (0·23)

7. There were positive and significant genetic correlations between casein and both fat and lactose, and that between the last two was almost significant.

8. No significant genetic correlations were found between yield and the chemical constituents, although figures of practical importance may exist which did not reach significance in an experiment of this size.

9. The practical significance of these results is discussed. It is concluded that selection for yield alone is not likely to cause any great decline in S.N.F. content of the milk, and that this could probably be held in check by paying some attention to the fat content of the milk.

Group N streptococci were found to cleave threonine to form acetaldehyde and glycine. Threonine aldolase, the enzyme catalysing this reaction, was found in all strains except Streptococcus cremoris Z8, an organism which had been shown previously to have a nutritional requirement for glycine. The enzyme was strongly inhibited by glycine and cysteine. The inhibition showed characteristics of allosteric inhibition and was pH-dependent. Inhibition by glycine, but not by cysteine, was highly specific. Analogues and derivatives of cysteine which contained a thiol group and a free amino group inhibited the activity of threonine aldolase. The presence of a carboxyl group was not necessary for inhibition. The cleavage of threonine by wholecell suspensions was stimulated by either an energy source to aid transport, or by rendering the cells permeable to substrate with oleate. Threonine did not appear to be degraded by enzymes other than threonine aldolase, as threonine dehydratase activity was low and NAD- and NADP-dependent threonine dehydrogenases were absent.

The effects of high pressure (HP) treatment (100–400 MPa at 10–60°C) on the solubility of milk protein concentrate (MPC) powders were tested. The solubility, measured at 20°C, of fresh MPC powders made with no HP treatment was 66%. It decreased by 10% when stored for 6 weeks at ambient temperature (∼20°C) and continued to decrease to less than 50% of its initial solubility after 12 months of storage. Of the combinations of pressure and heat used, a pressure of 200 MPa at 40°C applied to the concentrate before spray drying was found to be the most beneficial for improved solubility of MPC powders. This combination of pressure/heat improved the initial cold water solubility to 85%. The solubility was maintained at this level after 6 weeks storage at ambient temperature and 85% of the initial solubility was preserved after 12 months. The improved solubility of MPC powders on manufacture and on storage are attributed to an altered surface composition arising from an increased concentration of non-micellar casein in the milk due to HP treatment prior to drying. The improved solubility of high protein powders (95% protein) made from blends of sodium caseinate and whey protein isolate compared with MPC powders (∼85% protein) made from ultrafiltered/diafiltered milk confirmed the detrimental role of micellar casein on solubility. The results suggest that increasing the non-micellar casein content by HP treatment of milk or use of blends of sodium caseinate and whey proteins are strategies that may be used to obtain high protein milk powders with enhanced solubility.

While the effect of long-chain fatty acids on adipose tissue (AT) lipogenic activities has been described in non-lactating ruminants (Vernon, 1977), little is known about their effects on the mammary gland and the AT in lactating animals. However, in cows in mid lactation, duodenal rapeseed oil infusion decreased the rate of fatty acid (FA) synthesis in AT and increased milk yield of long-chain FA (18[ratio ]1, 18[ratio ]2 and 18[ratio ]3) and decreased medium-chain FA (14[ratio ]0 and 16[ratio ]0), suggesting a depressive effect of fat feeding on mammary lipid synthesis de novo (Chilliard et al. 1991). On the other hand, in goat species, the addition of vegetable lipids to the diet led to an increase in the milk fat content and yield (Chilliard et al. 2003) suggesting that the possible negative effect of long-chain FA on FA synthesis in the lactating mammary gland could be more than compensated by increasing the supply of FA brought to the mammary gland for milk synthesis. Elsewhere, AT from various anatomical sites are characterized by different FA composition in goat (Bas et al. 1987) together with different patterns of lipogenic gene expression in sheep (Barber et al. 2000). These results suggest that each AT site is characterized by a specific metabolism. However, in lactating ruminants, few data are available on the extent of expression and regulation of genes coding for lipogenic enzymes in AT. Therefore, the current study was performed in three lipogenic tissues of lactating goats, namely the mammary gland, an internal AT site (perirenal AT) and an external AT site (subcutaneous AT).

Intracellular peptidases of Lactobacillus helveticus may play a major role in the proteolysis of Swiss cheeses, provided that they are released through bacterial lysis. Experimental Swiss cheeses were manufactured on a small scale from thermized and microfiltered milk using as starters (in addition to Streptococcus thermophilus and Propionibacterium freudenreichii) one of two Lb. helveticus strains, ITGLH1 and ITGLH77, which undergo lysis to different extents in vitro. All the cheeses were biochemically identical after pressing. The viability of Lb. helveticus ITGLH1 and ITGLH77 decreased to a similar extent (96–98%) while in the cold room, but the concomitant release of intracellular lactate dehydrogenase in cheeses made with strain ITGLH1 was 5–7-fold that in cheeses made with ITGLH77. Protein profiles and immunoblot detection of the dipeptidase PepD confirmed a greater degree of lysis of the ITGLH1 strain. Free active peptidases were detected in aqueous extracts of cheese for both strains, and proteolysis occurred principally in the warm room. Reversed-phase HPLC revealed a more extensive peptide hydrolysis for ITGLH1, which was confirmed by the greater release of free NH2 groups (+33%) and free amino acids (+75%) compared with ITGLH77. As the intracellular peptidase activities of ITGLH1 and ITGLH77 have previously been shown to be similar, our results indicated that the extent of lysis of Lb. helveticus could have a direct impact on the degree of proteolysis in Swiss cheeses.

Colostral administration practices on dairy farms have significantly improved over the last 15–20 years resulting in prevalence of calves ingesting insufficient colostrum decreasing from 35–40% to 19%. Despite these improvements, the serum immunoglobulin G (IgG) concentration of ≥1000 g/dl and serum total protein (TP) concentrations of ≥5·2 g/dl are considered indicative of adequate transfer of immunity. We hypothesised that the current serum IgG concentrations of ≥1000 mg/dl is too low to indicate adequate transfer of colostral immunity on modern dairies. The objective of this study was to determine the serum IgG and TP concentrations indicating adequate transfer of passive immunity in dairy heifer calves. A cohort study of 1290 heifers from a calf raising facility for 48 dairy farms was performed. Heifers were assigned into strata based on serum IgG and TP concentrations. Mortality events were recorded for the heifers for 4 months. Interval likelihood ratios for mortality were calculated for heifers in each stratum of serum IgG or TP concentrations. Logistic regression to predict probability of mortality events was performed. Estimates of probability of survival were evaluated using survival analysis. Serum strata of ≤1500, 1501–2000 or >2500 were not significant predictors of mortality during the 120 d of rearing. Serum IgG concentration was not a significant predictor of hazard for mortality. In contrast to previous studies, serum IgG and TP concentrations of 2001–2500 mg/dl and 5·8–6·3 g/dl respectively, were considered optimum for indicating adequate passive transfer of colostral immunity in dairy calves based on the likelihood ratios. On dairies with optimum colostral feeding practices, serum IgG and TP concentrations of 2001–2500 mg/dl and 5·8–6·3 g/dl are recommended as endpoints to indicate adequate passive immunity in dairy calves.

Samples of liquid and solid fractions obtained by a commercial process from anhydrous milk fat of softening point (S.P.) 33·5–34°C had S.P. values of 22–23°C and 36–38°C respectively.

Determinations of fatty acid composition by GLC showed that unsaturated and short chain fatty acids were present in increased concentration in the liquid fraction (average 37·8 and 12·4% as compared with 35·1 and 10·8% in the original milk fat) and long chain saturated acids in the solid fraction (average 57·8 as compared with 53·8%). There was some concentration of carotene and vitamin A, and to a lesser extent of cholesterol, in the liquid fraction.

Crystallization and melting curves determined by a differential scanning calorimeter (DSC) showed that while the liquid fraction was completely melted at 25°C, the solid fraction contained an increased proportion of fat melting between 30 and 40°C. In addition to these high melting triglycerides the solid fraction also contained some 65% of fat in the liquid phase at the original crystallization temperature of 25°C.

Bovine mastitis is the most important infectious disease on dairy farms. Conventional antibiotic therapy is often unsatisfactory and alternative treatments are continually under investigation. Lactobacillus (Lb.) perolens CRL 1724 and Lactobacillus plantarum CRL 1716 were previously isolated from milk of dairy cows and selected according to their potential probiotic properties. In the present work the in-vitro capacity of Lactobacillus strains to adhere to bovine teat canal epithelial cells (BTCEC) and to inhibit and co-aggregate 14 mastitis-causing pathogens (MCPs) was investigated. The effect of Lb. perolens CRL 1724 after intramammary inoculation in lactating cows was evaluated through determination of clinical signs of mastitis, milk appearance, somatic cell counts and Lb. perolens CRL 1724 recovery from milk. Lb. perolens CRL 1724 was able to inhibit 12 of 14 MCPs (85·7%) in vitro, especially those considered to be major pathogens. In addition, Lb. perolens CRL 1724 co-aggregated with all of them. Lb. plantarum CRL 1716 was able to inhibit 7 of 14 MCPs (50%) in vitro and showed co-aggregation ability similar to Lb. perolens CRL 1724. Lb. perolens CRL 1724 showed a higher efficacy of adhesion to BTCEC (values of percentage of adhesion and adhesion index of 75% and 14·4, respectively) than Lb. plantarum CRL 1716 (37% and 7·4, respectively). Lb. perolens CRL 1724 was recovered from all mammary quarters and no clinical signs or teat damage were observed after the inoculation of 106 cfu/ml. The udders presented a normal aspect and there were no changes in the appearance of the milk. The results obtained will serve as the basis for further trials to evaluate the potential of Lb. perolens CRL 1724 to be included in a non-antibiotic formulation for the prevention of bovine mastitis.

Bifidogenic effect and stimulation of short chain fatty acid (SCFA) production by fractions of oligosaccharides with a DP⩾3 and Gal β(1-6) linkages synthesised from lactose or lactulose by Pectinex Ultra SP-L and Lactozym 3000 L HP G were evaluated in human faecal slurries. Results were compared with those obtained for the commercial oligosaccharide mixture Vivinal®-GOS. Quantitative real-time PCR showed that all galacto-oligosaccharide (GOS) fractions stimulated slightly higher bifidobacteria growth than lactose, lactulose and Vivinal®-GOS. GOS fractions promoted the production of total SCFA and acetic acid in a similar way to Vivinal®-GOS and more than glucose, lactose and lactulose. In conclusion, oligosaccharides derived from lactose and lactulose may represent a suitable alternative to lactulose for the design of new functional food ingredients.

The equipment tested (Fossomatic) is a fully automatic electronic instrument for counting the somatic cells in milk samples. The results are printed on a paper strip and displayed on a digital counter. The capacity is 180 samples/h.

Counting results obtained with the instrument have been compared with direct microscopic cell counts and particle counts by Coulter counter. The correlation coefficient with microscopic cell counts was 0·922–0·988, and with counts by the Coulter counter it was 0·916–0·992.

Reproducibility showed a coefficient of variation of 2–5 %. The carry-over between the samples was 2·0 %. Preserving milk samples with 0·05 % potassium dichromate did not influence cell counts, whereas freezing the samples reduced the counts 30 %.

Milk contains the alkaline proteinase plasmin and lysosomal proteinases; the significance of the latter is ill-defined. The objective of this study was to investigate composition and activities of several different proteolytic enzymes in milk samples of varying somatic cell count (SCC). Increasing milk SCC was correlated with increased plasmin, cathepsin D and cysteine protease activities, with concomitant increases in proteolysis in milk. Addition of plasmin inhibitors confirmed the heterogeneity of proteinase activities in milk, as urea-PAGE analysis of milk samples showed casein hydrolysis in milk after 7 d storage even in samples with inhibitors added; extent and heterogeneity of proteolysis was correlated with milk SCC. Rennet coagulation properties were not significantly correlated with SCC, or activities of measured enzymes. Milk of increasing SCC also exhibited decreased physical stability during incubation of milk at 37 °C. Pasteurized milk was more stable than raw milk, suggesting that the enzyme(s) or mechanisms leading to such instability are impaired by pasteurization. Overall, milk has a very heterogeneous proteolytic enzyme population, with a higher significance of non-plasmin enzymes, such as cathepsin D and cysteine proteinases, than perhaps previously recognised.

The free glucose concentration in the aqueous phase of samples of goat, sheep, cow, rat and rabbit milk was about 0·1–0·3 mn, while that in human milk was about 2 mM. During starvation the glucose concentration of goat milk fell considerably (by about 80 % in 2 d) in parallel with the decreased rate of lactose production. With rats fed ad lib., glucose concentration in the milk was greater at 12.00 h than at 18.00 h, when lactose synthesis has been shown to decrease. 3-O-Methyl-D-glucose injected into the goat mammary gland via the teat canal specifically entered the blood. These findings support the idea that glucose equilibrates across the apical membrane of mammary secretory cells, so that milk glucose concentrations reflect intracellular glucose concentrations.

The aim of the study was to identify the species of Enterobacteriaceae present in Montasio cheese and to assess their potential to produce biogenic amines. Plate count methods and an Enterobacterial Repetitive Intergenic Consensus Polymerase Chain Reaction (ERIC-PCR) approach, combined with 16S rDNA sequencing, were used to investigate the Enterobacteriaceae community present during the cheesemaking and ripening of 6 batches of Montasio cheese. Additionally, the potential decarboxylation abilities of selected bacterial isolates were qualitatively and quantitatively assessed against tyrosine, histidine, ornithine and lysine. The most predominant species detected during cheese manufacturing and ripening were Enterobacter cloacae, Escherichia coli and Hafnia alvei. The non-limiting physico-chemical conditions (pH, NaCl% and aw) during ripening were probably the cause of the presence of detectable levels of Enterobacteriaceae up to 120 d of ripening. The HPLC test showed that cadaverine and putrescine were the amines produced in higher amounts by almost all isolates, indicating that the presence of these amines in cheese can be linked to the presence of high counts of Enterobacteriaceae. 44 isolates produced low amounts of histamine (<300 ppm), and four isolates produced more than 1000 ppm of this amine. Only 9 isolates, belonging to the species Citrobacter freundii, Esch. coli and Raoultella ornithinolytica, appeared to produce tyramine. These data provided new information regarding the decarboxylase activity of some Enterobacteriaceae species, including Pantoea agglomerans, Esch. fergusonii and R. ornithinolytica.

In milk samples from 549 cows of the breeds Danish Jersey, Red Danish Dairy Cattle (RDM), and Black and White Danish Dairy Cattle (SDM) the genetic polymorphisms of the αs1, β and K-cascin and β-lactoglobulin (β-Lg) loci were determined by isoelectric focusing in agarose gels. The results of the screening were comparcd with results obtained by Larsen & Thymann (1966). In addition, the genetic linkage of the three casein loci was studied, and the association between milk protein genotypes and yields in first and second lactations of milk, fat and protein were investigated.

The distribution of genotypes of all four milk protein Systems was different from breed to breed.

For Jersey cows, significant differences in the gene frequencies from the results of the 1966 investigation were found for αs1 and K-casein and β-Lg. For SDM cows a change in the K-casein frequency had occurred whereas for RDM cows no changes were found.

Linkage between some of the casein loci was found within ail three breeds. For the RDM breed the possible linkage between αs1-casein and the other caseins could not be tested bccause nearly ail thc cows were homozygous for the αs1-cascin-B genotypes.

β-Casein genotypes were associated with yield parameters in ail breeds. The A2A2 genotype of this protein gave higher yields of milk, fat, and protein in the second lactation than thc A1A1 genotype.

Prevalence of mastitis pathogens in milk samples from dairy cows and heifers was studied over a period of 1 year (Aug 2005–Aug 2006) in ten dairy herds in Germany. Milk samples (n=8240) were collected from heifers without clinical mastitis at parturition (n=6915), from primiparous cows with clinical mastitis (n=751) and from older cows with clinical mastitis (n=574). Coagulase negative staphylococci (CNS) were the predominant group of bacteria isolated (46·8% of samples) from clinically healthy quarters of primiparous cows around parturition, followed by streptococci (12·6%), coliforms (4·7%) and Staphylococcus aureus (4·0%). Thirty-three percent of samples were negative on culture (range on farm level, 12·0–46·4%). In cases of clinical mastitis in primiparous and older cows, streptococci were the predominant finding (32·1 and 39·2%) followed by CNS (27·4 and 16·4%), coliforms (10·3 and 13·1%) and Staph. aureus (10·0 and 11·7%). Negative results were obtained from 21·3% (range, 0·0–30·6%) and 19·5% (range, 0·0–32·6%) of these samples. Results indicated substantial differences in the prevalence of pathogens among herds. There was a positive within-herd correlation between the monthly prevalences for Streptococcus dysgalactiae between the three groups of samples. This correlation was also found between clinical samples of primiparous and older cows for Staph. aureus. These correlations were not found for the other pathogens. Besides herd, prevalence of pathogens was influenced by parity, type of sample and season.