Data from 33 feeding trials, conducted on lactating riverine buffaloes from different institutes across India, were subjected to multiple regression analysis to derive nutritional requirements for dry matter (DM), total digestible nutrients (TDN), crude protein (CP) and digestible crude protein (DCP) for maintenance, milk production and body weight gain. Maintenance requirements for DM, TDN, CP and DCP were 59.9, 35.3, 5.43 and 3.14 g/kgW0.75, respectively; corresponding requirements for producing 1 kg 6% FCM were 688, 406, 90.3 and 55.2 g and for 1 g gain in body weight were 3.37, 1.97, 0.327 and 0.23 g. Regression equations had high R2 values (0.61, 0.66, 0.84 and 0.68 for prediction of DM, TDN, CP and DCP, respectively) and the equations (F-value) as well as coefficients were highly significant (P<0.001). Regressed values were used to derive feeding standards. Derived values matched well with the actual intake versus performance of animals under diverse feeding conditions. New standards predicted requirements and intake of nutrients for different production levels better than existing feeding standards. Because they are based on a more thorough analysis of data, the new feeding standards will be appropriate for use widely in India.

Forty lactating Comisana ewes were either exposed to or protected from solar radiation and fed either in the morning or afternoon during summer in a Mediterranean climate. Individual milk samples were taken on days 7, 21 and 42 of the study period to determine fatty acid composition by gas chromatography. Exposure to solar radiation resulted in higher proportions of short-chain and saturated fatty acids in milk, primarily because of increased contents of caproic, capric, lauric, myristic and stearic acids (by 3–18%), and decreased contents of oleic, linoleic and linolenic acids (by 2–9%). As a consequence, the long to short chain and the unsaturated to saturated fatty acid ratios were significantly higher by 4 and 13% respectively in the milk of the protected ewes compared with that of the exposed animals. Provision of shade also led to an increase in the 18[ratio ]0+18[ratio ]1 to 16[ratio ]0 ratio, and to a decrease in the 12[ratio ]0+14[ratio ]0+16[ratio ]0 fatty acid group, which are regarded as reliable indexes of the nutritional property of dietary fat in reducing cholesterol levels in human plasma. Feeding time had little impact on milk fat. Our findings suggest that high ambient temperature may markedly modify the lipid composition of ewe milk and that provision of shade, but not feeding management, can improve the milk fatty acid profile in dairy sheep raised in hot climates.

Changes in urokinase-plasminogen activator (u-PA) and u-PA receptor (u-PAR) expression at the protein and mRNA level in resting neutrophils and in neutrophils activated by phorbol myristate acetate (PMA) were examined. Low amounts of u-PA were found intracellularly or membrane-bound in resting neutrophils. However, incubation of resting neutrophils with purified exogenous u-PA (10 IU/ml) revealed extensive binding of u-PA to cell membranes. Excess amino-terminal fragment of the u-PA molecule, a proteolytically inactive fragment of u-PA (amino acids 1–135) blocked binding of exogenous u-PA to the cell membrane. These results, collectively, indicate that the binding of u-PA is specific and that resting neutrophils have unoccupied u-PA receptors on their cell membrane. Addition of PMA led to an increase (P<0.01) in total cell-associated, membrane-bound u-PA activity and u-PA mRNA expression by bovine neutrophils. In contrast, PMA increased u-PAR mRNA levels but this was accompanied by a decrease (2.5-fold; P<0.01) in free, unoccupied u-PA binding sites. No significant effects on total cell-associated or membrane-bound u-PA were found when neutrophils were treated with 4-phorbol 12,13 didecanoate, a phorbol ester that does not activate protein kinase C (PKC). Furthermore, addition of 1-(5-isoquinolinesylphonyl)-2-methylpiperazine dihydrochloride (H-7), a potent PKC inhibitor, blocked the effect of PMA on total cell-associated u-PA activity. Thus, PKC plays a role in the modulation of u-PA and u-PAR by PMA in bovine neutrophils.

The amount of protein synthesis translational elongation factor 2 (eEF-2) was estimated employing diphtheria toxin-dependent ADP-ribosylation in samples prepared from small amounts of tissue from mammary gland, skeletal muscle and liver from lactating dairy cows. A very high level of ADP-ribosylatable eEF-2 was found in mammary gland, amounting to 20-times the level found in liver and 50-times the level found in skeletal muscle. This obviously reflects the high protein synthesis activity in mammary tissue. To our knowledge, similar high activities have previously been reported only for cancer cells. A close linear relationship was found between the amount of diphtheria-toxin catalysed ADP-ribosylated eEF-2 and protein and casein output in milk from cows in late lactation. This strongly suggests that eEF-2 may be a limiting factor in milk protein synthesis.

Sixty samples of raw goat milk intended for Caprino cheese-making were collected from ten farms in the Bergamo area over a 6-month period. Analyses of main microbial groups, somatic cell count (SCC) and pH were performed to determine the effect of origin (farm) and lactation period (April – September) on microbial composition and the incidence of pathogens in milk. Overall mean values were: standard plate count (SPC), 5.0×104 cfu/ml; yeasts, 2.5×102 cfu/ml; coliforms, 9.1×102 cfu/ml; Escherichia coli, 2.9 cells/ml; enterococci, 1.1×102 cfu/ml; lactococci, 3.4×103 cfu/ml; lactobacilli, 3.0×103 cfu/ml; halotolerant bacteria, 8.2×103 cfu/ml; spores of mesophilic aerobic bacteria, 11 cfu/ml; SSC, 9.9×105 cells/ml; pH, 6.63. Moulds and spores of sulphite-reducing clostridia were found intermittently. Neither Salmonella spp. nor Listeria monocytogenes was detected, while Esch. coli O157[ratio ]H7 was isolated from one milk sample (an incidence of 1.7%). Staphylococcus aureus was discovered at a level >102 cfu/ml in 26 samples (43%) with an overall mean of 1.2×103 cfu/ml, whereas coagulase-negative staphylococci were found in 54 samples (90%) with an overall mean of 1.3×103 cfu/ml. Of Staph. aureus strains, 23% proved to be enterotoxinogenic with a prevalence of enterotoxin C producers. Staph. caprae was the coagulase-negative species most frequently isolated; none of the coagulase-negative staphylococci strains synthesized any of the enterotoxins tested for.

Sample source was the major factor affecting the microbial composition of goat milk: significant differences (P<0.01) were observed among samples from different farms for SPC, coliforms, lactococci, lactobacilli and halotolerant bacteria. Period of lactation had a significant effect (P<0.025) on SCC and pH. SPC correlated well with coliforms, lactococci and lactobacilli; SSC did not reveal positive interactions with any microbial groups or pH.

The reason for the reported difference in spoilage behaviour of skim and whole pasteurised milks was investigated. The rates of growth of psychrotrophic bacteria were not significantly different in the two milks and the bacterial types, all pseudomonads, present at spoilage were also similar. However, when representative spoilage organisms were cultured into freshly pasteurised skim and whole milks, skim milks exhibited predominantly bitter flavours while whole milk showed mostly sour flavours. The different spoilage behaviours can be largely explained by greater proteolysis in skim milk than in whole milk, caused by higher production of protease and greater susceptibility of the protein to protease attack. In addition, lipolysis in whole milk, caused by the substantial quantities of lipase produced by spoilage pseudomonads in this milk, also contributes to the different flavours produced during cold storage of these milk types.

The effect of different parameters (pH, ultrasonic amplitude and pressure) on the resistance to heat and manothermosonication (MTS) treatments of heat resistant lipase and protease produced by Pseudomonas fluorescens B52 and NCDO 2085, respectively, were studied. Lipase B52 thermoresistance decreases with an increase of pH. However, inactivation by MTS seems to be pH independent. There were only slight increases in the MTS efficiency when increasing pressure at UHT temperatures and the effect of amplitude was different depending on treatment temperature. Protease NCDO 2085, which was very resistant to MTS at 30 °C, was very sensitive to MTS at 76 °C. Increases in applied pressure had no effect on MTS efficiency at 140 °C and its inactivation by MTS was almost temperature independent between 76–109 °C. Data obtained are compared with previous published data and inactivation mechanisms are discussed.

A homo-tetrameric ∼160-kDa cystathionine γ-lyase was purified to homogeneity from Lactobacillus reuteri DSM 20016 by four chromatographic steps. The activity was pyridoxal-5′-phosphate dependent and the enzyme catalyzed the α,γ-elimination reaction of L-cystathionine, producing L-cysteine, ammonia and α-ketobutyrate. The enzyme was active towards a range of amino acids and amino acid derivatives, including methionine. The pH and temperature optima were found to be 8.0 and 35 °C, respectively. Isoelectric pH (pI) was ∼5.0 as determined by two-dimensional electrophoresis. Sensitivity to chemical inhibitors was typical of lactococcal cystathionine γ- and β-lyases, except it was inhibited by sulphydryl reagents. The N-terminal sequence was MKFNTQLIHGGNSED, which had 100% homology with cystathionine β-lyase of Lb. reuteri 104R (Accession Number CAC05298). Lb. reuteri DSM 20016, together with 10 other strains of non-starter lactic acid bacteria, was used as adjunct starter in the production of miniature Canestrato Pugliese-like cheeses. After 40 d ripening, the water-soluble extract of the cheeses with added Lactobacillus fermentum DT41 and Lb. reuteri DSM 20016 contained the highest enzyme activities on cystathionine and methionine substrates. Determinations of methanethiol, dimethyl sulphide, dimethyl disulphide and dimethyl trisulphide in the miniature cheeses confirmed the findings of enzyme activities.

The aspartic proteinase, chymosin (EC 3.4.23.4) is the principal milk clotting enzyme used in cheese production and is one of the principal proteolytic agents involved in cheese ripening. Varietal differences in chymosin activity, due to factors such as cheese cooking temperature, fundamentally influence cheese characteristics. Furthermore, much chymosin is lost in whey, and further processing of this by-product may require efficient inactivation of this enzyme, with minimal effects on whey proteins. In the first part of this study, the thermal inactivation kinetics of Maxiren 15 (a recombinant chymosin preparation) were studied in skim milk ultrafiltration permeate, whole milk whey and skim milk whey. Inactivation of chymosin in these systems (at pH 6.64) followed first order kinetics with a D45.5 value of 100±21 min and a z-value of 5.9±0.3 °C. D-Values increased linearly with decreasing pH from 6.64 to 6.2, while z-values decreased as pH decreased from 6.64 to 6.4, but were similar at pH 6.4 and 6.2. Subsequent determination of chymosin activity during manufacture of Cheddar and Swiss-type cheese showed good correlations between predicted and experimental values for thermal inactivation of chymosin in whey. However, both types of cheese curd exhibited relatively constant residual chymosin activity throughout manufacture, despite the higher cooking temperature applied in the manufacture of Swiss cheese. Electrophoretic analysis of slurries made from Cheddar and Swiss cheese indicated decreased proteolysis due to chymosin activity during storage of the Swiss cheese slurry, but hydrolysis of sodium caseinate by coagulant extracted from both cheese types indicated similar levels of residual chymosin activity. This may suggest that some form of conformational change other than irreversible thermal denaturation of chymisin takes place in cheese curd during cooking, or that some other physico-chemical difference between Swiss and Cheddar cheese controls the activity of chymosin during ripening.

Experimental Camembert cheeses were made to investigate the effects on browning of the following factors: inoculation with Yarrowia lipolytica, the use of Penicillium candidum strains with different proteolytic activity, the addition of tyrosine, and the addition of Mn2+, thus leading to 16 different variants of cheese. Two physical colour parameters were used to describe browning, depending on the location in the cheeses: a whiteness index for the outside browning (mould mycelium), and a brownness index for the inside browning (surface of the cheese body). Mn2+ promoted a significant increase of browning at both locations, whereas Yar. lipolytica had the opposite effect. Outside browning was significantly more intense when using the Pen. candidum strain with higher proteolytic activity. A significant interaction was found between Yar. lipolytica and Pen. candidum. The yeast had no effect in combination with a low proteolytic strain of Pen. candidum, but significantly reduced proteolysis and browning in combination with a high proteolytic strain of Pen. candidum. We further confirmed that both strains of Pen. candidum were able to produce brown pigments from tyrosine, and thus both are presumably responsible for the browning activity in this type of cheese.

The action of bile on bifidobacteria was studied by measuring changes in zeta potential, hydrophobicity and adherence to enterocyte-like cells in vitro. Highly hydrophobic strains shocked with bile displayed more negative zeta potential values and a decrease in adherence. When a non-hydrophobic non-adherent strain (CIDCA 5324) was shocked with bile, an increase in hydrophobicity was observed. However, no changes of zeta potential or adherence properties were apparent. The action of the bile components was different from the action of whole bile. Cholate and deoxycholate produced a decrease in the negativity of zeta potential values of all strains studied whereas taurocholate displayed a shift in zeta potential of hydrophobic strains to more negative values, thus explaining the decrease in the autoaggregation by charge repulsion. However, the decrease in zeta potential caused by cholate and deoxycholate did not increase autoaggregation in a hydrophobic non-adherent strain (CIDCA 531). This suggests that other forces are contributing to autoaggregation.

The therapeutic and preventive effects of yogurt and lactic acid bacteria on diseases such as cancer, infection and gastrointestinal disorders are well documented. The aim of this research was to study the effects of different doses of yogurt addition after milk re-nutrition diet, on the recovery of the intestinal barrier and mucosal immune function. Experiments were performed on groups of mice, malnourished and re-nourished with milk during 7 d, and mice with diet supplemented with yogurt for 2, 5 and 7 consecutive d. Nutritional parameters such as weight gain, serum total protein, and the number of IgA, IgM and IgG B cells of the small intestine were determined. We also quantified intraepithelial leukocytes, mastocytes and goblet cells, and performed structural and ultrastructural studies on the small intestine. We observed that 5 d of yogurt feeding was the optimal dose for improving gut barrier function and mucosal immune system in a malnutrition model. This effect was not observed with milk re-nutrition. Although the results were better for 5 d of yogurt, addition for 7 d also showed beneficial effects. Yogurt feeding in our model did not impair any gut functions. These results suggest that yogurt addition after a re-nutrition diet gives better recovery of intestinal function than the re-nutrition diet usually recommended. Although these results were obtained in an animal model, they indicate that consumption of yogurt by malnourished children might accelerate the restoration of gut function.

Monensin sodium is widely used to manipulate ruminal fermentation (Bergen & Bates, 1984), with the aim of increasing energy supply to the animal. Monensin has been most widely used in diets of beef cattle and young growing dairy cattle, particularly in confinement management systems, where the ionophore can be mixed directly into the ration. Accurate daily dosing of grazing ruminants with small quantities of rumen modifiers, such as ionophores, proved particularly labour-intensive and expensive until the development of the controlled-release capsule (CRC; Laby et al. 1984). Subsequently a commercially available CRC containing monensin sodium was developed (Elanco Animal Health, Cambridge, NZ) facilitating its use in grazing animals. It has also been tested as a method of reducing the incidence of bloat in lactating dairy cows grazing pastures containing legumes.

Since its identification by Devirese et al. (1983), Staphylococcus caprae has been frequently isolated from udder halves of goats with subclinical intramammary infection (IMI) (Contreras et al. 1995; Poutrel et al. 1996). Several different studies have demonstrated the capacity of Staph. caprae to persist throughout lactation (Poutrel et al. 1996; Contreras et al. 1997; Bedidi-Madani et al. 1998a) and during the dry period (Poutrel, 1984). Bedidi-Madani et al. (1998a) found that isolates of Staph. caprae from udders of uninfected goats express potential virulence factors (different exoproteins and slime); other researchers have occasionally isolated it from clinical cases of goat mastitis (Deinhofer & Pernthaner, 1995), or from human clinical specimens associated with bone and joint infection (Vandenesh et al. 1995; Shuttleworth et al. 1997). In spite of these forms of pathogenicity, some studies have not shown significant increases in somatic cell count (SCC) of milk from goat udder halves infected by Staph. caprae (Deinhofer & Pernthaner, 1995; Poutrel et al. 1996). Moreover, there is a lack of information on the effect of infection by this mammary pathogen on milk yield and composition. Knowledge of repercussions for hygiene and production would be useful in mastitis control programmes in view of the high frequency of isolation (higher than 22% of the staphylococci isolated) of Staph. caprae in dairy goat herds (Contreras et al. 1995; Poutrel et al. 1996). The aim of the present investigation was to ascertain the significance of persistent subclinical IMI by Staph. caprae on SCC, milk yield and physical-chemical parameters of Murciano-Granadina goat milk at mid-lactation.

Milk is known to provide not only excellent nutrition but also protection against infection in mammalian neonates. Whey proteins, such as immunoglobulins, lactoferrin, lysozyme and lactoperoxidase, are well known antimicrobial components in milk. Håkansson et al. (1995) demonstrated that some multimeric human α-lactalbumin derivatives induced apoptosis in immature and tumour cells. Yoo et al. (1997) found that lactoferricin, which had been isolated from peptic lactoferrin as an antimicrobial peptide, induced apoptosis in tumour and immature mammalian cells. Apoptotic cell death is considered to be an important physiological function in the development and homeostasis of multicellular organisms and to be a highly controlled means of eliminating dangerous, damaged or unnecessary cells without causing tissue damage. Therefore, these findings suggest that whey proteins may have beneficial health effects by restricting specific cell population in the body.

Antimutagenic properties of various dietary components have been investigated in many studies, in order to find compounds protective against the various steps of carcinogenesis. Indeed, cancer is now the principal cause of death in industrialised countries and treatment is still difficult. Thus, research in prevention is of extreme importance. While, the etiology of most cancers has an environmental component, colon or breast cancers are highly linked to dietary habits (up to 50% of colorectal cancers are due to diet; Riboli et al. 1996). In this context, Boutron et al. (1996) showed inverse correlation between consumption of dairy products and colorectal cancer in France.