The objective was to promote a reduction in the prevalence of Staphylococcus aureus and Streptococcus dysgalactiae after 2 years of selective dry cow therapies and teat dipping/external teat sealant implementation. Three different dry cow treatments, one long-acting and two short-acting penicillin-based products were tested at herd level together with a negative control teat dipping group, an iodine teat dipping group and DryFlex™, an external teat sealant. The regimes were independently randomly allocated to 178 dairy herds. Yearly bacteriological quarter milk samples were collected from all cows at the beginning of the trial, after 1 year and 2 years. At herd level, a total of 15% of the herds showed no Staphylococcus aureus isolates after 2 years, compared with 5% at the beginning. The distribution of Streptococcus dysgalactiae infected herds remained the same after 2 years. At cow level, there were no significant differences in the reduction of Staph. aureus between the different dry cow therapies and teat dipping regimes. But there was a significant reduction of Str. dysgalactiae in the iodine teat dipping group compared with Dryflex™ and the negative control group. The proportional rate of Staph. aureus positive quarters was reduced from 65·9% to 54·9% after 2 years. As for Str. dysgalactiae, an increase was observed from 14·2% to 15·2%.

A detailed investigation was undertaken to determine the effects of four single starter strains, Lactococcus lactis subsp. lactis 303, Lc. lactis subsp. cremoris HP, Lc. lactis subsp. cremoris AM2, and Lactobacillus helveticus DPC4571 on the proteolytic, lipolytic and sensory characteristics of Cheddar cheese. Cheeses produced using the highly autolytic starters 4571 and AM2 positively impacted on flavour development, whereas cheeses produced from the poorly autolytic starters 303 and HP developed off-flavours. Starter selection impacted significantly on the proteolytic and sensory characteristics of the resulting Cheddar cheeses. It appeared that the autolytic and/or lipolytic properties of starter strains also influenced lipolysis, however lipolysis appeared to be limited due to a possible lack of availability or access to suitable milk fat substrates over ripening. The impact of lipolysis on the sensory characteristics of Cheddar cheese was unclear, possibly due to minimal differences in the extent of lipolysis between the cheeses at the end of ripening. As anticipated seasonal milk supply influenced both proteolysis and lipolysis in Cheddar cheese. The contribution of non-starter lactic acid bacteria towards proteolysis and lipolysis over the first 8 months of Cheddar cheese ripening was negligible.

Yak milk casein derived from Qula, a traditional Tibetan acid curd cheese, was hydrolyzed by six commercially available proteases (Trypsin, Pepsin, Alcalase, Flavourzyme, Papain and Neutrase). These hydrolysates were assayed for their inhibitory activity of Angiotensin-I-converting enzyme (ACE). The hydrolysates obtained by Neutrase from Bacillus amyloliquefaciens showed the highest ACE inhibitory activity. The IC50 value of Neutrase-hydrolysate was 0·38 mg/ml. The hydrolysate obtained by Neutrase was further separated by consecutive ultra-filtration with 10 kDa and then with 6 kDa molecular weight cut-offs into different permeated parts and fractionated by gel filtration chromatography with a Sephadex G-25 column. The active fraction was subjected to RP-HPLC, in which five peaks were purified and identified. Amino acid sequence analysis confirmed that the peptides and origins were as follows: YQKFPQY (αs2-CN; f89–95), LPQNIPPL (β-CN; f70–77), SKVLPVPQK (β-CN; f168–176), LPYPYY (κ-CN; f56–61) and FLPYPYY (κ-CN; f55–61). Their amino acid sequences matched well with those of known bioactive peptides from bovine casein. The results indicated that yak milk casein could be a resource to generate antihypertensive peptides and be used as multifunctional active ingredients for many value-added functional foods as well as a traditional food protein.

Effects of ventilation rate and of dietary protein level in an intensive dairy sheep system on the features of Canestrato Pugliese cheese were studied. Cheeses were manufactured from the bulk milk obtained from ewes subjected to four different experimental treatments: (1) low dietary crude protein (CP) of 13% in dry matter (DM) at a low ventilation rate (23·5 m3/h per ewe) (LPLV); (2) low dietary CP at a moderate ventilation rate (47 m3/h per ewe) (LPMV), (3) moderate dietary CP of 16% DM at a low ventilation rate (MPLV); and (4) moderate dietary CP dietary at a moderate ventilation rate (MPMV). Bulk milk and cheeses (at 1, 15, 45 and 90 d of ripening) were analysed for chemical composition, N fractions and plasmin-plasminogen activities. The pH 4·6-soluble and insoluble N fractions were analysed by urea-PAGE. Bulk milk from ewes receiving the low CP diet displayed higher casein and lower urea contents. Ewes subjected to the low ventilation rate displayed a higher plasminogen activity in milk, whereas no differences emerged among treatments in the conversion of zymogen to plasmin. During ripening the plasmin-plasminogen system in cheese did not display significant changes across treatments. At 90 d of ripening, the cheese produced with milk from ewes receiving the low CP diet and exposed to the low ventilation rate displayed more markedly stained bands in the area of γ-caseins in pH 4·6-insoluble N fraction. As from 15 d of ripening, the pH 4·6-soluble fraction showed a greater number of bands in the LPMV and MPMV than in the LPLV and MPLV cheeses. The results showed that Canestrato Pugliese cheese manufactured with milk from ewes fed the low CP diet and exposed to the moderate ventilation rate was characterized by higher protein and casein content in the fresh cheese and by a greater proteolysis after 90 d of ripening.

Reducing the dietary cation-anion difference (DCAD) has been shown to be an effective means of preventing parturient paresis in confinement systems where cows are offered a total mixed ration containing DCAD-reducing mineral compounds (anionic salts). Such a supplementation strategy is not possible in cows being group fed forages precalving, and little is known about the effect of supplementing these cows with large amounts of anionic salts twice daily.

Eight non-lactating, pregnant Holstein-Friesian cows were allocated to two levels of DCAD (−20 and +18 meq/100 g DM) for 24 d, with an intensive Ca balance undertaken in metabolism stalls following a 2-week acclimatization to diet. The basal diet was 3 kg DM of crushed barley and 7 kg DM of pasture-hay. Urine and faeces were collected separately, weighed daily for 5 d and analysed for Ca content. Urinary Ca, creatinine and hydroxyproline concentration and plasma Ca concentration were determined during the period of the balance study. The diurnal pattern in urine and rumen pH was determined over 2 d. Decreasing DCAD reduced (P<0·001) the pH of urine, and increased (P<0·05) Ca absorption. Plasma Ca concentration was not affected by DCAD, and DCAD did not affect the output of urinary hydroxyproline, a marker of bone resorption. Twice-daily supplementation of anionic salts was sufficient to reduce the pH of blood and increase gastrointestinal Ca absorption. There was no diurnal variation in the pH of urine, suggesting that time of sampling to determine efficacy of DCAD in reducing systemic pH was not important.

A longitudinal study involving 73 primiparous (PP) and 47 multiparous (MP) Holstein cows was conducted over an 8-month period to assess the associations between locomotion score (LCS) and milk production, dry matter intake (DMI), feeding behaviour, and number of visits to an automatic milking system (AMS). Twice weekly, all cows were locomotion scored (scale 1–5) by the same observer. Individual eating behaviour and individual feed consumption at each cow visit to the feed troughs, individual milk production, the time of milking, and the number of milkings for each cow were recorded for the day of locomotion scoring and the day before and after. Dependent variables, such as milk yield, DMI, etc. were modelled using a mixed-effects model with parity, LCS, days in milk (DIM), the exponential of −0·05 DIM, and the interaction between parity and LCS, as fixed effects and random intercepts and random slopes for the linear and the exponential of −0·05DIM effects within cow. LCS did not affect time of attendance at feed troughs, but affected the location that cows occupied in the feed troughs. The time devoted to eating and DMI decreased with increasing LCS. Milk production decreased with LCS>3. The number of daily visits to the AMS also decreased with increasing LCS. The cows with high LCS were fetched more often than the cows with low LCS. Overall, PP cows were more sensitive to the effects of increasing LCS than were MP cows. The decrease in milk production observed with increasing LCS seemed to be affected similarly by the decrease in DMI and by the decrease in number of daily visits to the AMS. A further economic loss generated by lame cows with AMS will be associated with the additional labour needed to fetch them.

The DGAT1 gene encodes a microsomal enzyme that catalyses the only committed step in triacylglycerol synthesis by joining diacylglycerol and fatty acyl coenzyme A. In cattle, a K232A substitution in the DGAT1 molecule has a significant effect on enzyme activity and milk fat content. The prominent role of this gene in lipid metabolism led us to undertake the structural characterization of DGAT1 in goats. In this way, we have sequenced a 1552 bp fragment of the goat DGAT1 cDNA, which encompasses most of the coding sequence (from exon 1 to 17), and a genomic fragment covering exons 12 to 17. Multiple alignment of the goat DGAT1 sequences revealed the existence of a single nucleotide polymorphism (SNP) involving a T to C substitution at intron 16. We optimized a primer extension based genotyping method that allowed us to determine that the C variant is a minority allele with frequencies ranging from 0·062 (Murciano-Granadina) to 0·109 (Malagueña). This SNP, although not expected to have any functional effect, might be useful as a genetic marker in association studies to detect additional DGAT1 polymorphisms which might influence fat milk content and other traits of economic interest.

The aim was to determine whether administration of melatonin would alter the yield and composition of milk from grazing dairy cows in summer. Twelve sets of spring-calving identical twin Friesian cows were used in the experiment. In late-November (late spring), one twin from each set was given slow-release melatonin implants behind the ears (108 mg melatonin/cow). Two further implantations occurred at 4-weekly intervals to maintain increased circulating concentrations of melatonin for 12 weeks. The other twin served as a control. Milk yield and composition were measured twice prior to treatment and then four times over the following 12 weeks. Concentrations of melatonin, prolactin and insulin-like growth factor 1 (IGF-1) were measured in blood plasma twice before treatment and then either seven (melatonin and prolactin) or three (IGF-1) further times during the experiment. Management procedures for all cows were similar and cows grazed a daily pasture allowance of approximately 30 kg DM/cow as their sole feed source. In melatonin-treated cows there was a decrease in mean concentrations of prolactin in plasma, but concentrations of IGF-1 did not change. Melatonin reduced milk yield by 6 weeks after treatment and by the end of the 12-week experimental period milk yield in melatonin-treated cows had fallen by 23%. Melatonin also reduced concentrations of lactose in milk, but increased concentrations of fat, protein and casein, changes that were broadly similar to those that occur in late lactation in seasonally calving dairy cows. Thus, the results suggest that some of the variation in the volume and quality of milk throughout the season in New Zealand dairy systems may be due to changes in photoperiod mediated by increased concentrations of plasma melatonin in association with decreased concentrations of plasma prolactin.

To evaluate the bioactivity of bovine milk from different stages of lactation on human intestinal tissue, a human fetal small intestinal cell line was used as a model system. Milk samples representing six stages of lactation: days 1, 2–3, 6–7 and weeks 12 and 24 after parturition, 1 week before drying off, and milk-like secretion from two stages of the dry period: 7 weeks and 3–4 weeks before expected calving, were collected from 64 Holstein Friesian cows. The whey fraction of the milk or milk-like secretion was added to the culture medium in concentrations ranging from 0·078% to 10%. The growth-promoting activity of whey was measured by determining the incorporation of [3H]thymidine into DNA for the last 24 h of the culture period. Whey fractions from all six stages of lactation stimulated growth of intestinal cells. The growth-promoting activity of colostrum or milk significantly decreased within the first week after calving. The growth-promoting activity in mature milk increased gradually during lactation to reach a level significantly higher than that obtained with colostrum. The growth-promoting activity of whey from milk-like secretion collected after drying off was lower than that of colostrum. Whey from different stages of lactation contained significantly different concentrations of TGF-β1 (0·5–27 ng/ml) and TGF-β2 (12–1219 ng/ml). However, neither the differences in TGF-β1 and TGF-β2, nor the differences in IGF-I and IGF-binding proteins could fully explain the differences in growth-promoting activity of colostrums or milk from different stages of lactation, suggesting that other factors were also involved. The present study showed that bovine milk contained a number of biologically active components that affected growth and development of human intestinal tissue. The results showed that the growth-promoting activity of colostrum and milk was dependent on the stage of lactation in accordance with previous results obtained with mammary epithelial cells. The changes in growth-promoting activity with stage of lactation were probably related to changes in concentrations of several growth factors.

Data from 350 herds enrolled in the Norwegian Dairy Herd Recording System (NDHRS) were used to investigate the associations between the first two cow-milk somatic cell counts (SCC) test-days’ results after calving or the three last SCC test-days prior to drying off in the first lactation and the hazard ratio (HR) of clinical mastitis (CM) during the remaining first or the subsequent second lactation respectively. Altogether, 9519 first lactations and 6046 second lactations were included. Cox regression analyses adjusted for herd frailty effect were used. In the first lactation, SCC>40000 cells/ml on the first or second test-day was significantly associated with an increased risk of a CM event in the remaining first lactation. HR, compared with 10000 cells/ml, increased from 1·6 (1·4) for SCC of 40000–60000 cells/ml to 6·9 (4·2) for SCC >800000 cells/ml, when using the first (second) SCC test-day in the first lactation. Cows with a geometric mean of the three last SCC test-days between 50000 and 100000 cells/ml and between 401000 and 800000 cells/ml in the first lactation had HR of CM during the second lactation of 1·3 and 2·8 respectively compared with a reference group of 10000–20000 cells/ml. If a CM episode in the first lactation occurred, the HR for having a CM event during the second lactation was 1·5. There was a significant frailty effect which disappeared if the incidence rate of CM at herd level was included in the model.

Propolis is a natural product collected by honeybee workers. The product was tested for its antifungal effect against Aspergillus versicolor ATCC 12996 as well as biosynthesis of sterigmatocystin during ripening of Egyptian Ras cheese. The use of different concentrations of aqueous propolis extract 250, 500 and 1000 part per million (ppm) on the cheese surface was investigated. Mould growth and toxin production were completely inhibited at the highest concentration 1000 ppm, while the lower concentrations exhibited definite fungistatic activity during 90 days of ripening. Control cheese demonstrated that the amount of sterigmatocystin produced was proportional to the growth of Asp. versicolor during three months of ripening. It could be concluded that propolis concentration of 1000 ppm could prevent mould growth and sterigmatocystin production in Ras cheese. The economic as well as the public health importance of propolis as a natural preservative in cheese manufacture is discussed.

Bovine Somatotropin (bST) is a peptide hormone secreted by the anterior pituitary gland and its recombinant form (rbST) is used for artificially boosting milk yield in cows. Identification of rbST is difficult in that there is little difference from the pituitary bST (pbST). In this work, we further studied the possibility of immunologically discriminating between rbST and pbST. With this purpose, we produced mouse monoclonal antibodies using, as antigen, a peptide mimicking the N-terminus of rbST from Monsanto (rbST-M) conjugated to keyhole limpet haemocyanin (KLH) and polyclonal antibodies in rabbits immunized with the whole bST or rbST-M. Hence, we developed a sandwich ELISA with the obtained antibodies for detection and quantification of bST in serum and compared its performance on the two worldwide commercialized rbSTs: rbST-M and rbST from LG Life Science (rbST-LG). The lowest detection limit of the assay was 0·05 ng/ml for rbST-M, 0·10 ng/ml for rbST-LG and 0·15 ng/ml for pbST. Furthermore, the assay showed the capability to amplify the signal in the presence of rbSTs, recognizing more efficiently rbST-M and rbST-LG than pbST (ECn pbST/ECn rbST: 3 and 1·6 respectively). Its employment for measuring bST levels in sera from bovines administered with rbST LG allowed us to detect exceptional values due to the treatment itself and probably further increased as a consequence of the higher affinity for rbSTs of our monoclonal antibody.

The objective was to compare the prevalence of subclinical mastitis (SM) and of udder pathogens in 60 Swiss organic (OP) and 60 conventional production systems (CP). Cows (n=970) were studied for SM prevalence and udder pathogens at median 31 d and 102 d post partum. Cows showing a [ges ]1+ positive California Mastitis Test (CMT) in at least one quarter were considered to have SM. Cow-level prevalences of SM for visits at 31 d and 102 d post partum (39% and 40% in OP and 34% and 35% in CP) were similar, but quarter-level prevalences of SM were higher (P<0·02) in OP than CP (15% and 18% in OP and 12% and 15% in CP). Median somatic cell counts in milk at 31 d post partum were higher (P<0·05) in OP than CP cows (43000 and 28000 cells/ml, respectively), but were similar at 102 d post partum in OP and CP cows (45000 and 38000 cells/ml, respectively). In milk samples from quarters showing a CMT reaction [ges ]2+ the prevalences of coagulase negative staphylococci were lower (P<0·05) at 102 d post partum, whereas prevalences of non-agalactiae streptococci were higher (P<0·05) in OP than in CP cows at 31 d and 102 d post partum. In conclusion, under Swiss conditions, subclinical mastitis is a greater problem in organic than in conventional production systems, but differences are not marked.

Our aim was to develop an explant model to define more precisely the early response of bovine mammary epithelial cells to infection. Therefore we investigated the mRNA expression encoding for some soluble immunological factors in lipopolysaccharide (LPS)-treated bovine mammary gland explants. Explants were taken out from the mammary gland of eight lactating cows after slaughter then incubated with LPS (10 μg/ml) for 6 h. The mRNA expression of α-lactalbumin (α-la), various cytokines, tumour necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, IL-8, and two immunoglobulin receptors, the neonatal Fc receptor (FcRn) and polymeric immunoglobulin receptor (pIGR), were assessed with qPCR before and after 3 h and 6 h of LPS challenge. Both immunoglobulin receptors and α-la increased at 3 h then recovered their initial level at 6 h whereas IL-1β, IL-6 and IL-8 increased only after 6 h (P<0·05). Surprisingly, TNF-α transcripts did not show any regulation in response to the LPS treatment. We nevertheless concluded that our model was valid to examine the short-term response of mammary epithelial cell challenged with LPS.

Latex immune agglutination method with a multipurpose auto-analyser (the automated latex assay) was validated for determination of bovine lactoferrin (BLF) in various dairy products. Reproducibility-within-laboratory (intermediate precision) due to day for infant formula, UHT milk and yogurt supplemented with BLF at 50 mg/100 g for infant formula and UHT milk, and at 100 mg/100 g for yogurt were 1·62 to 3·10 mg/100 g. Reproducibility-within-laboratory due to analysis (morning, noon, and evening) for raw milk was 1·59 mg/100 g. Trueness, accuracy in determining known amounts added for BLF-supplemented dairy products was −4·7 to −2·0 mg/100 g. BLF concentration in raw milks was 20·3 to 21·8 mg/100 g. Although interference by the matrixes took place in infant formula and raw milk, BLF assays were accurately carried out by 2000-fold dilution or standard-addition method. Automated latex assay for BLF is simple, rapid, precise and accurate enough for a routine method in various dairy products containing BLF.

Age thickening is a well known phenomenon within the dairy industry, where the apparent viscosity of concentrated dairy solutions increases with storage time under low shear. The objective of the paper is to develop new tools to analyse several aspects of age thickening, including the increases in shear thinning, the degree of thixotropy and the rate of structural build up with time, as well as irreversible structural changes and gelation. The build up of the dispersed phase structure during storage, was captured by a snap shot technique, where concentrated milk samples were stored, and aliquots were taken at periodic intervals and subjected to shear sweep tests to collect flow curves. Rheological modelling using the Herschel-Bulkley model showed an increase in yield stress and consistency, indicating a build up in structure during age thickening. The trend in apparent viscosity with storage time showed two-phases, a slow and steady increase followed by a sharp rise, which could be used to identify the onset of firm gelation. The extrapolated viscosity at infinite shear rate increased with storage time, indicating the build up in irreversible structural components during age thickening.

This work investigated the combined effect of solids content, heating and storage temperatures on the rheological behaviour of reconstituted whole milk concentrates. The powder was reconstituted at 35 °C in a custom built recombination rig to various total solids content (TS) from 10–48% TS. The concentrates were then heated to 45–85 °C and stored at the heating temperature. The rheological behaviour shifter from Newtonian behaviour (below 30% TS) to power law (below 40% TS), with the yield stress observed from 40% TS upwards and time-dependent behaviour was noticed above 44% TS. Higher heating temperatures tend to promote non-Newtonian behaviour at lower solids content. The viscosity-solid content curve showed an exponential relationship, while the viscosity-temperature curve exhibited a minimum at 65–75 °C above 46% TS. During age thickening, the yield stress and the consistency coefficient increased, while the flow behaviour index decreased with storage time. This indicated that the milk concentrates deviated away from Newtonian behaviour during age thickening.

Dubert-Ferrandon A, Niranjan K and Grandison AS 2006 A novel technique for differentiation of proteins in the development of acid gel structure from control and heat treated milk using confocal scanning laser microscopy. Journal of Dairy Research73(4) 423–430

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