A global process for the production of goat milk hydrolysates enriched in angiotensin converting enzyme (ACE) inhibitory peptides was proposed. Firstly, the protein fractions (caseins and whey proteins) were separated by ultrafiltration through a 0·14 μm ceramic membrane. The casein fraction obtained in the retentate stream of the above filtration step was subsequently hydrolysed with a combination of subtilisin and trypsin. After 3 h of reaction, the hydrolysate produced presented an IC50 of 218·50 μg/ml, which represent a relatively high ACE inhibitory activity. Finally, this hydrolysate was filtered through a 50 kDa ceramic membrane until reaching a volume reduction factor of 3. The permeate produced presented an improvement of more than 30% in the ACE inhibitory activity. In contrast, the retentate was concentrated in larger and inactive peptides which led to a decrease of more than 80% in its inhibitory activity. The process suggested in this work was suitable to obtain a potent ACE inhibitory activity product able to be incorporated into food formulas intended to control or lower blood pressure. Moreover, the liquid product could be easily stabilised by spray dried if it would be necessary.

The effects of use of adjunct cultures (Lactobacillus helveticus and Lb. casei) and ripening temperatures (6 or 12 °C) on proteolysis and angiotensin-I converting enzyme (ACE)-inhibitory activity in white-brined cheeses were investigated during 120 d ripening. Proteolysis was monitored by urea-polyacrylamide gel electrophoresis (urea-PAGE) and reversed phase-HPLC (RP-HPLC) of water-insoluble and -soluble fractions of the cheeses, respectively. Urea-PAGE patterns of the samples revealed that the intensities of the bands representing casein fractions decreased in the experimental cheeses, being more pronounced in the cheeses made with adjunct cultures. Similarly, peptide profiles and the concentrations of individual and total free amino acids were influenced by both the adjunct cultures and ripening temperatures. The ACE-inhibitory activity of the water-soluble extracts of the cheeses were higher in the cheeses made using adjunct cultures (especially Lb. helveticus) and ripened at 12 °C. The ACE-inhibitory activity did not decrease during ripening. The contribution of Lb. helveticus to the development of proteolysis and ACE-inhibitory peptide activities were higher than that of Lb. casei. To conclude, the use of Lb. helveticus as adjunct culture in white-brined cheese and ripening at 12 °C would be recommended to obtain white-brined cheese with high ACE-I-inhibitory peptides activity and higher levels of preoteolysis.

The length of the dry period in commercial dairy production is under close scrutiny. While the main concern is the composition and volume of milk produced, the evaluation of colostrum quality under these new paradigms has suggested a decline in IgG concentrations, while some reports indicate no change. Colostrum quality has been defined as an adequate concentration (>50 mg/ml) of immunoglobulin in the secretions to provide the newborn with maximal disease resistance. We investigated the appearance of IgG in mammary pre- and post partum secretions in cows without a dry period (continuously milked, Dry0) and compared the secretions with cows that experienced a dry period of 60 d (Dry60). Blood was collected during the experimental period and plasma analysed for progesterone (P4) and prolactin (Prl). Approximately −6 d relative to parturition, the Dry0 animals exhibited increased concentration of IgG in their secretions to an average of ∼35 mg/ml that remained rather constant through subsequent pregnancy and following parturition. Dry0 cows were producing an average IgG concentration in parturition colostrum of 44·2±17·6 mg/ml that was not different than that of controls (66·86±16·8 mg/ml). However, Dry0 cows exhibited high variation, different peak times (day) of IgG concentration including times that occurred both pre and post parturition. IgG mass of the Dry0 cows remained rather constant pre- and post partum and did not show the same declining mass following parturition that was shown for the Dry60 cows. The change in plasma P4 and Prl were shown to have no timing effect on colostrum IgG concentration.

The influence of different amounts of oilseed cake (rapeseed and sunflower) on animal production parameters and fatty acid (FA) concentrations of the milk was studied in a Latxa dairy sheep experimental flock, both in winter (50% oilcakes; indoor feeding) and in spring (30% oilcakes; part-time grazing). The two different levels of the oilcakes tested did not affect animal production parameters or milk yield. Milk fat content and the fat/protein ratio decreased significantly with 30 and 50% sunflower cake. Yet, fat/protein ratio values were within the range for cheesemaking. Both levels of either type of oilcake tested significantly increased the concentrations of nutritionally interesting FA (CLA isomer C18:2cis-9, trans-11, vaccenic, oleic, and total unsaturated FA), while simultaneously decreasing the concentration of atherogenic FA. The atherogenicity indexes of milks from ewes fed 50 or 30% of either oilcake were significantly lower than those of their corresponding control. The use of cakes in winter increased the concentration of nutritionally interesting FA to the values obtained with part-time grazing.

Water soluble autolysate of yeast, usually utilised for microbial growth support, was used as additive in yoghurt fermentation. The yeast extract (YE) resulted in a decrease of fermentation time by 21% to reach a pH of 4·6. However, the YE resulted in unacceptable flavour and taste. By size exclusion chromatography, a fraction of the YE was obtained that could account for the observed 21% decrease in fermentation time. The fraction contained molecules of low molecular weight, consisting of minerals, free amino acids and peptides. The acceleration of the yoghurt fermentation was ascribed to the short peptides in the fraction. It is proposed that the application of this extract in industrial yoghurt manufacture would result in savings for both the industry and the consumer.

Extended duration of clinical mastitis (CM) treatment has been advocated, although results showing its higher efficacy compared with standard treatment are difficult to compare and seem conflicting. In a non-blinded, positively controlled clinical trial with systematic allocation, the efficacy of a standard, 1·5-d cefquinome treatment (ST), and an extended, 5-d intramammary cefquinome treatment (ET) were evaluated. The latter is frequently performed in cows with persistent high somatic cell count (SCC), expecting a better cure. Therefore, cows with CM immediately preceded by at least two consecutive monthly elevated SCC >200 000 cells/ml, were studied. The primary efficacy criteria were bacteriological cure (BC) and clinical cure (CC), while SCC cure was considered a secondary criterion of cure. Least square means of overall BC were not different after ET (79%, n=206) compared with ST (72%, n=203). ET, as compared with ST, improved BC of CM when caused by streptococci, specifically Streptococcus uberis. At day 1·5, only 13% of quarters showed CC, increasing significantly towards 60% at day 5, and 99% at day 14 and at day 21. No significant difference in CC was present between treatment groups. Overall SCC cure was low (22%) and not significantly different between treatment groups, but significantly higher for cases due to enterobacteriacae compared with staphylococci. In conclusion, ET with cefquinome of CM in cows with a persistent high SCC seems to be only indicated when caused by streptococci, mainly Str. uberis but shows no advantage when no information on bacteriological causes of mastitis is available. In our data, absence of CC directly after ST was not related to eventual BC.

Domestic cattle have limited cardiopulmonary reserve for their body size and oxygen requirements. Therefore, it is plausible that impaired alveolar-arterial gas exchange may be detrimental to energetically expensive traits such as milk production which, like all aerobic processes, requires oxygen. The degree of alveolar-arterial oxygen transfer impairment can be determined by estimating the alveolar-arterial oxygen (A-a O2) pressure gradient from arterial blood-gas tensions. The degree of oxygen transfer impairment is proportional to the A-a O2 pressure gradient: the higher the A-a O2 pressure gradient the less oxygen is transferred to the blood for a given ventilation rate. In this study two cohorts of Holstein-Friesian heifers were followed on one northern Colorado dairy farm. Arterial blood-gas analyses were performed up to 9 d post-calving. Heifers were grouped into quartiles based on A-a O2 pressure gradient so that relative comparisons could be made. Heifers in the lowest (Q1) and highest (Q4) quartile had the least and greatest impairment of alveolar-arterial oxygen transfer, respectively. We hypothesised that milk yield over 60 d would be greatest for heifers in Q1 and would decrease with quartile increments. Hyperventilation, as indicated by hypocapnia, was notable. Despite hypoxia, haematocrit was low. Alveolar-arterial O2 pressure gradient was associated with milk production (P=0·03) when controlling for cohort, treatment for disease and calving difficulty score. Heifers in Q1 produced 1992 kg (95% CI=1858, 2127 kg) of milk when controlling for all other variables. Relative to heifers in Q1, heifers in Q2, Q3 and Q4 produced 130 kg (95% CI=313, −52 kg; P=0·45), 285 kg (95% CI=474, 96 kg; P=0·004) and 169 kg (95% CI=395, −57 kg; P=0.14) less milk, respectively. In conclusion, efficacy of alveolar-arterial oxygen transfer was associated with milk yield in dairy heifers on one farm at moderate altitude.

Instruments for on-farm determination of colostrum quality such as refractometers and densimeters are increasingly used in dairy farms. The colour of colostrum is also supposed to reflect its quality. A paler or mature milk-like colour is associated with a lower colostrum value in terms of its general composition compared with a more yellowish and darker colour. The objective of this study was to investigate the relationships between colour measurement of colostrum using the CIELAB colour space (CIE L*=from white to black, a*=from red to green, b*=from yellow to blue, chroma value G=visual perceived colourfulness) and its composition. Dairy cow colostrum samples (n=117) obtained at 4·7±1·5 h after parturition were analysed for immunoglobulin G (IgG) by ELISA and for fat, protein and lactose by infrared spectroscopy. For colour measurements, a calibrated spectrophotometer was used. At a cut-off value of 50 mg IgG/ml, colour measurement had a sensitivity of 50·0%, a specificity of 49·5%, and a negative predictive value of 87·9%. Colostral IgG concentration was not correlated with the chroma value G, but with relative lightness L*. While milk fat content showed a relationship to the parameters L*, a*, b* and G from the colour measurement, milk protein content was not correlated with a*, but with L*, b*, and G. Lactose concentration in colostrum showed only a relationship with b* and G. In conclusion, parameters of the colour measurement showed clear relationships to colostral IgG, fat, protein and lactose concentration in dairy cows. Implementation of colour measuring devices in automatic milking systems and milking parlours might be a potential instrument to access colostrum quality as well as detecting abnormal milk.

We used a mouse model of pathogenic (Staphylococcus aureus) and non-pathogenic (teat sealing) mammary inflammation to investigate mRNA expression of several inflammatory cytokines and acute phase proteins (APP) in mammary tissue and liver, and the appearance of some of these factors in plasma and milk. The expression levels of IL1β and TNFα were markedly up-regulated in Staph. aureus-inoculated mammary tissue at 72 h, whilst IL6 was up-regulated to a lesser extent in a way which was not confined to the inoculated glands. APP expression was up-regulated at 48 and 72 h in both Staph. aureus-inoculated and teat-sealed mammary glands. These differences between cytokine and APP expression provide additional support for the contention that APPs are produced within the mammary tissue itself during inflammation, rather than in associated immune cells. We propose that measurement of cytokines and APP in combination might provide a tool for diagnostic discrimination between mastitis caused by pathogenic invasion and milk accumulation, and hence allow for better targeting of antibiotic therapy. In comparison with mammary expression, expression of cytokines in liver tissue was up-regulated to a similar or lesser extent, whilst expression of APP was up-regulated to a much greater extent. The first appearance of increased cytokine and APP concentrations in plasma and of milk amyloid A (MAA) in milk occurred in advance of the measurable up-regulation of expression, hence their origin cannot be stated with certainty.

Pecorino cheeses were produced using two different manufacturing procedures (either from raw milk or from milk previously subject to a thermisation at 65 °C for 10 s with the addition of starter culture) using the milk obtained from three groups of ewes fed three different concentrates: (1) control, (2) control enriched with 100 g/kg of extruded linseed, and (3) control enriched with 200 g/kg of extruded linseed. All concentrates were administered with alfalfa hay ad libitum. The extruded linseed-enriched diets increased the concentration of n-3 fatty acids, conjugated linoleic acid, and monounsaturated and polyunsaturated acids in raw milk and thermised milk cheese and decreased the concentration of saturated fatty acids. Changes in the manufacturing procedure did not influence the fatty acid composition of the cheeses but markedly influenced their sensory properties. In particular, heat-treating the milk and adding a starter reduced the differences in cheese odour, flavour and toughness induced by the diet when raw milk was used. Cheeses made with thermised milk and the addition of starter culture were more uniform and obtained a higher score in the preference test compared with the corresponding raw milk cheeses. An appropriate manufacturing process, therefore, can be a possible strategy to obtain cheeses with improved health-promoting properties and an unaltered acceptance level by consumers.

Sampling approaches following the dairy chain, including microbiological hygiene status of critical processing steps and physicochemical parameters, contribute to our understanding of how Staphylococcus aureus contamination risks can be minimised. Such a sampling approach was adopted in this study, together with rapid culture-independent quantification of Staph. aureus to supplement standard microbiological methods. A regional cheese production chain, involving 18 farms, was sampled on two separate occasions. Overall, 51·4% of bulk milk samples were found to be Staph. aureus positive, most of them (34·3%) at the limit of culture-based detection. Staph. aureus positive samples >100 cfu/ml were recorded in 17·1% of bulk milk samples collected mainly during the sampling in November. A higher number of Staph. aureus positive bulk milk samples (94·3%) were detected after applying the culture-independent approach. A concentration effect of Staph. aureus was observed during curd processing. Staph. aureus were not consistently detectable with cultural methods during the late ripening phase, but >100 Staph. aureus cell equivalents (CE)/ml or g were quantifiable by the culture-independent approach until the end of ripening. Enterotoxin gene PCR and pulsed-field gel electrophoresis (PFGE) typing provided evidence that livestock adapted strains of Staph. aureus mostly dominate the post processing level and substantiates the belief that animal hygiene plays a pivotal role in minimising the risk of Staph. aureus associated contamination in cheese making. Therefore, the actual data strongly support the need for additional sampling activities and recording of physicochemical parameters during semi-hard cheese-making and cheese ripening, to estimate the risk of Staph. aureus contamination before consumption.

The possibilities of using high quality pastures in conjunction with total mixed ration (TMR) during the grazing season have been examined. An experiment with sixteen Holstein cows blocked and randomly assigned to four treatments in a factorial arrangement was conducted in order to evaluate the influence of grazing time of day (day or night) and type of silage (maize or Italian ryegrass) included in the TMR of dairy cows grazing 12 h daily on milk yield, composition and fatty acid profile. The silage type had no effect on the dry matter intake, milk yield and fat and protein proportions. However, cows grazing during the night ate more grass than cows grazing during the day (8·53 vs. 5·65 kg DM/d; P<0·05). No differences were seen between grazing-time with respect to milk production, fat and protein contents. However, the proportion of polyunsaturated fatty acid was higher in milk of dairy cows grazing at night-time than grazing at day-time, especially 18:2n-6 (2·37 vs. 2·12 g/100 g FA respectively, P<0·05) and 18:2cis9trans11 (2·08 vs. 1·74 g/100 g FA respectively, P<0·05).

In order to protect peroral β-galactosidase from being degraded and hydrolyse milk lactose efficiently in the environments of gastrointestinal tract, a double-capsule delivery system composed of enteric-coated capsule and polylactic acid (PLA) nanocapsules (NCs) was developed for encapsulation of β-galactosidase. β-galactosidase-loaded PLA NCs in the size range of 100–200 nm were prepared by a modified w1/o/w2 technique. During the encapsulation process, dichloromethane/ethyl acetate (1 : 1, v/v) as the solvent composition, high-pressure homogenisation (150 bar, 3 min) as the second emulsification method and polyvinyl alcohol or Poloxamer 188 as a stabiliser in the inner phase could efficiently improve the activity retention of β-galactosidase (>90%). Subsequently, the prepared NCs were freeze-dried and filled in a hydroxypropyl methylcellulose phthalate (HP55)-coated capsule. In vitro results revealed that the HP55-coated capsule remained intact in the simulated gastric fluid and efficiently protected the nested β-galactosidase from acidic denaturation. Under the simulated intestinal condition, the enteric coating dissolved rapidly and released the β-galactosidase-loaded PLA NCs, which exhibited greater stability against enzymatic degradation and higher hydrolysis ratio (∼100%) towards milk lactose than the free β-galactosidase. These results suggest that this double-capsule delivery system represents promising candidate for efficient lactose hydrolysis in the gastrointestinal tract.

Milk yield and milk qualitative parameters were evaluated in Lacaune ewes on a diet supplemented with extruded linseed, verbascoside and vitamin E. A 98 d-trial was conducted on 44 ewes and started 40±2 d post partum. The animals were divided into four homogeneous groups of eleven animals each; one control group (CON) without extruded linseed and dietary supplements, and the diet of the other three experimental groups was enhanced with extruded linseed (L group), extruded linseed-verbascoside (LVB group), and extruded linseed-verbascoside-vitamin E (LVBE group). All animals individually received an isoenergetic diet, consisting of 700 g concentrated feed and meadow hay ad libitum. Body weight, body condition score, milk yield and milk qualitative parameters were assessed. LVB and LVBE groups resulted in a significant improvement (P<0·05) in milk yield due to the verbascoside supplementation. The extruded linseed supplementation L, LVB and LVBE groups produced a milk fat increase and a better milk fatty acid profile in terms of a higher monounsaturated fatty acid (MUFA), polyunsaturated fatty acid (PUFA) and conjugated linoleic acid (CLA) content and a reduced saturated fatty acid (SFA) content, a lower n-6/n-3 ratio and atherogenic and thrombogenic index. The dietary verbascoside supplementation in the LVB and LVBE group resulted in a better milk quality due to the low cholesterol level and higher vitamin A and E contents, in addition to an increased oxidative stability highlighted by the lower thiobarbituric acid reactive substances (TBARS) level. Thus, the addition of extruded linseed and verbascoside supplements improved milk yield and quality both from a chemical and nutritive point of view.

We studied the effects of changes in the milking routine (lack or presence of 30-s prestimulation, 0 or 1, 2 or 4-min delay between preparation and cluster attachment) and environmental perturbation (unusual loud sounds capable of frightening animals just after stall entry or during the course of milking) on milk removal and milking-related behaviour in dairy dromedary camels. A 30-s prestimulation decreased incidence of bimodal milk flow curves and increased occurrence of the best milk ejection patterns with higher milk flow but had limited effect on milk production in our well-trained animals within a good machine milking setting. However, unusual sounds heard from the beginning of milking or even after milk ejection caused inhibition or disruption of milk removal and modification of camels’ behaviour. Milk ejection was significantly delayed (1·58±0·17 min), residual milk increased over 40% of total milk yield and average and peak milk flow rates were significantly lowered when unusual noises were heard from the beginning of milking. These environmental perturbations increased signs of vigilance and the number of attempts to escape the milking parlour. Delaying cluster attachment for over 1 min after the end of udder preparation caused serious milk losses. Up to 62% of total milk was withheld in the udder when the delay reached 4 min. Average and peak milk flow rates also decreased significantly with delayed milking. Signs of vigilance and attempts to escape from the milking parlour appeared when camels waited for over 2 min. After a 4-min delay, camels showed signs of acute stress. Defaecation prior to milk ejection (solid faeces) and rumination during milking can be used to assess camels’ milk ejection during milking. Animal welfare and milking efficiency can be ensured when camels are pre-stimulated, milked in calm conditions and with cluster attachment within a maximum of a 1-min delay after stimulation