Time spent lying by lactating Holstein-Friesian cows of varying body condition scores (BCS) and milk yield was measured using an animal activity monitor. A 3-week average BCS was calculated for each cow; and in total, 84 cows were selected with 28 cows each among three BCS categories (Thin: BCS<2·75; Moderate: 2·75⩾BCS<3·25; Heavy: BCS⩾3·25) and two stage of lactation categories (<150 days in milk or >150 days in milk). Cows were kept in two management systems: parlour/freestall (n=60) or automated milking system/freestall (n=24). Behaviour was recorded for 5·3±0·1 d for each cow. Production levels were considered using a 28-d rolling average of daily milk production. Cows that exhibited clinical lameness before or during the observation period were excluded from analyses. For cows exhibiting oestrus, the day prior to, day of, and day following breeding were removed. The final analysis included 77 cows (408 d of observation). A mixed model was fitted to describe average daily hours spent lying. Results demonstrated that lying time increased as days in milk (DIM) increased (P=0·05). Variables that were tested but not significant (P>0·05) were BCS category, parity category (1 or ⩾2) and 28-d rolling average daily milk production. Although a numerical trend for increasing hours spent lying with increasing BCS was observed, after accounting for other factors in the mixed model, BCS did not significantly impact lying time. Continued investigation of these management factors that impact lying time and bouts, using new technologies, more cows, and more herds will help dairy owners better manage facilities and cow movements to optimize this essential behaviour.

Two greatly related Lactobacillus plantarum bacteriophages (named FAGK1 and FAGK2) were isolated from Kefir grains of different origins. Both phages belonged to the Siphoviridae family (morphotype B1) and showed similar dimensions for head and tail sizes. The host range of the two phages, using 36 strains as potential host strains, differed only in the phage reactivity against one of them. The phages showed latent periods of 30 min, burst periods of 80±10 min and burst size values of 11·0±1·0 PFU per infected cell as mean value. Identical DNA restriction patterns were obtained for both phages with PvuI, SalI, HindIII and MluI. The viral DNA apparently did not present extremes cos and the structural protein patterns presented four major bands (32·9, 35·7, 43·0 and 66·2 kDa). This study reports the first isolation of bacteriophages of Lb. plantarum from Kefir grains and adds further knowledge regarding the complex microbial community of this fermented milk.

To investigate the relationships between milk flow traits and udder health status in primiparous cows, 74 primiparous Holstein cows were randomly selected in 5 herds and monitored monthly throughout the whole lactation. A total of 2902 quarter milk samples were collected for bacteriological analyses and the determination of lysozyme, N-acetyl-β-glucosaminidase (NAGase) and somatic cell count (SCC). Milk flow curves of the whole udder of each cow were registered with continuous electronic milk flow meters. Teat conditions and teat thickness changes during milking were assessed monthly. Quarters, udders and cows were classified as healthy, latent, inflamed and subclinical depending on SCC and the results of bacteriological analyses. Lysozyme in milk, teat apex score and teat thickness change did not vary with udder health status while NAGase in milk significantly increased as udder health status worsened (P<0·001). Milk production (P<0·001) and time of plateau phase (P<0·05) were significantly lower in subclinical cows in comparison with the others. Animals with a high frequency of bimodal curves in the first 100 days in milk showed the worst udder health status during the whole lactation (P<0·01). Moreover, cows classified as subclinical in the first 3 months of lactation had higher peak milk flow than healthy cows (3·81 v. 3·48 kg/min; P<0·05) and shorter duration of plateau phase, expressed both as minutes and as percentage of time of milk flow (pTPL; P<0·001). Multivariate logistic analysis showed udder health status to be associated with duration of plateau phase, time of milk flow, bimodality and duration of overmilking phase. With short time of plateau phase (pTPL <25%), short time of milk flow (<5 min), presence of bimodality and long overmilking phase (>0·8 min) there was an increased risk of poor udder health status. These milk flow traits can be predictive indicators of udder health status; time of plateau phase, expressed as percentage of time of milk flow, can also be a useful parameter for animal selection.

Two feeding systems trials were carried out to determine the antioxidant activity of soft goats' milk cheeses, and also to evaluate the presence of bioactive polyphenolic compounds. Two groups (A and B), each one with 20 (BW 50±5 kg) French Alpine goats were employed. All animals had between 70 to 80 milking days and were milked once a day. Group A included daily grazing on shruby rangeland vegetation, and group B was kept in full indoor confinement. Thirty kg of goats' milk from each group were collected; 15 kg from each were pasteurized and the remaining 15 kg were not pasteurized (raw), resulting in four treatments (two feeding system×two milk treatments). Soft goat's cheese antioxidant activity can be modified by the animals feeding system; grazing management represents a better option than indoor feeding to produce a healthy profile of bioactive compounds; providing an increase of total polyphenol, hydroxycinnamic acids and flavonoid concentrations. Pasteurization did have a significant effect on these metabolites, and diminished total polyphenol concentration. More research is needed to elucidate the potential of soft goat's cheese as a functional food.

Retinol-binding protein 4 (RBP4) is a plasma protein involved in retinol transportation, and recent evidence in rodents suggests that RBP4 is also a metabolic regulator that modifies insulin sensitivity. To assess how RBP4 levels are regulated in ruminants, we determined the RBP4 concentrations in bovine plasma and milk using Western blot analysis. Plasma RBP4 levels in non-pregnant non-lactating (control) cows were around 45 μg/ml, which were sustained during 60-h fasting, but decreased significantly 4 h after lipopolysaccharide (LPS) administration. Basal plasma retinol concentration was around 30 μg/dl, but this decreased to approximately one-third and one-half of these values during fasting and 8 h after LPS challenge, respectively. Plasma RBP4 and retinol levels in cows 3–6 d before parturition were comparable to those of the controls. However, on the day of parturition both were significantly decreased and had returned to basal levels by two weeks after calving. Interestingly, RBP4 was clearly detected in colostrum (16·4±5·6 μg/ml) but was only faintly detected in milk from cows at 7 d and 15 d after calving. Retinol concentrations in colostrum were almost 10-fold higher than those in plasma, while those in milk were comparable to those in plasma. These results suggest that RBP4 and retinol levels are independently regulated under physiological and pathophysiological conditions and that RBP4, like retinol, is transferred from maternal stores to calves through colostrum.

The aim of this study was to assess the concentration of penicillin G in mammary tissue and secretion of dry heifers following systemic administration of penethamate hydriodide. Six dairy heifers in late gestation received a single intramuscular injection of 10 g penethamate hydriodide and were sacrificed 24, 48 or 144 h after treatment. Penicillin G concentrations were measured in mammary tissue and secretion samples using HPLC. Penicillin G was detected in the udder of two animals euthanized at 24 h (mammary tissue and secretion) and at 48 h post treatment (mammary secretion only) after administration at concentrations still close to or above MIC90 values reported for the pathogens associated with heifer mastitis. Antibiotic concentration shortly after administration will have been substantially higher indicating a potential for systemic treatment with penethamate hydriodide to control prepartum intramammary infections in heifers without the disadvantages of local therapy such as teat contamination or risk of trauma for the administrator.

Insulin-like growth factor 2 (IGF2) is considered to be a regulator of post-natal growth and differentiation of the mammary gland. In the present work, associations of two single nucleotide polymorphisms in the bovine IGF2 gene with milk production traits were studied in dairy Holstein-Friesian cows: the already described g.8656C>T transition in exon 2 (RFLP-BsrI) and the newly found g.24507G>T transversion in exon 10 (RFLP-HaeIII), found by sequencing 273-bp exon 10 of the IGF2 gene in six individuals. Associations were analysed individually and in combination with the multi-trait repeatability test-day animal model. The CT/GT haplotype appeared to be associated with most of the milk traits studied (differences were significant at P⩽0·001). The most frequent CT/GG haplotype seemed inferior to others in fat and protein content and daily yield of fat and protein but superior (together with the TT/GG genotype) when the daily milk yield is considered.

The stability of camel α-lactalbumin (α-la) against heat denaturation was measured, using circular dichroism (CD) and fluorescence spectroscopy, as well as differential scanning calorimetry (DSC). The experiments were performed in the presence of saturating concentrations of calcium as well as in the presence of EDTA, yielding to the apo form of α-la. The change in heat capacity (ΔCp) suggests a greater contribution of hydrophobic interactions to the stability of holo camel α-la than in its bovine counterpart. Overall the results obtained in this study suggest a greater stability of camel α-la than the bovine protein in both holo and apo states. Also CD experiments showed similar secondary structure for camel and bovine α-la and secondary structure of camel α-la was better preserved than that of bovine α-la during heat denaturation. The differences in thermal stability between the proteins from two species can be primarily ascribed to the difference in the quantity of hydrophobic interactions involved in their folding.

This study explains how active coating can serve to prolong the shelf life of Fior di latte cheese. The active coating was prepared by dissolving, in two sodium alginic acid solutions (5 and 8% w/v), different concentrations of lysozyme (0·25, 0·50 and 1·00 mg ml−1)+50 mm of Ethylene-Diamine Tetraacetic Acid (EDTA). Samples of Fior di latte cheese packaged in brine and active brine (lysozyme+EDTA, at the above concentrations) were also used as controls. The quality decay of the Fior di latte cheese stored at 10°C was assessed by monitoring the viable cell concentration of the main spoilage microorganism, as well as its sensory quality (i.e., external appearance, consistency, colour and flavour). The concentration of rod-or coccus-shaped Lactic Acid Bacteria (LAB) was also monitored to assess the effect of the proposed packaging strategies on the flora type of Fior di latte cheese. The results show that an increase in the shelf life equal to 104% was recorded for the coated samples, compared with controls packaged in brine without active compounds. This shelf life increase is slightly lower than that recorded with samples packaged in the active brine (151%), as a result of a more pronounced microbial proliferation; however, the coating could be a better packaging solution for the reduced weight of tray.

Casein genes have been proved to have an influence on milk properties, and are in addition appropriate for phylogeny studies. A large number of casein polymorphisms exist in goats, making their analysis quite complex. The four casein loci were analyzed by molecular techniques for genetic polymorphism detection in the two dairy goat breeds Bunte Deutsche Edelziege (BDE; n=96), Weiße Deutsche Edelziege (WDE; n=91), and the meat goat breed Buren (n=75). Of the 35 analyzed alleles, 18 were found in BDE, and 17 in Buren goats and WDE. In addition, a new allele was identified at the CSN1S1 locus in the BDE, showing a frequency of 0·05. This variant, named CSN1S1*A′, is characterized by a t→c transversion in intron 9. Linkage disequilibrium was found at the casein haplotype in all three breeds. A total of 30 haplotypes showed frequencies higher than 0·01. In the Buren breed only one haplotype showed a frequency higher than 0·1. The ancestral haplotype B-A-A-B (in the order: CSN1S1-CSN2-CSN1S2-CSN3) occurred in all three breeds, showing a very high frequency (>0·8) in the Buren.

Milk is the normal secretion of the mammary gland, practically free of colostrum and obtained by the complete milking of one or more healthy animals. Mastitis is an inflammatory process of the mammary gland and it may cause alterations in the milk. The present work aimed to verify whether it is possible, by means of the counts of microorganism in the bulk raw milk in four selective culture media, to establish a correlation with the occurrence of mastitis and therefore, to monitor this disease in bovine dairy herds. The following selective culture media were used: KF Streptococcus Agar, Edwards Agar, Baird-Parker Agar, Blood Agar plus potassium tellurite. Spearman's correlation coefficient was calculated in order to compare the occurrence of mastitis (percentage) in each herd with respective selective culture media counts of microorganisms in bulk raw milk. Thirty-six possibilities were analysed (Tamis and CMT-positive rates were compared with the log-transformed count in four selective culture media) and there was a negative correlation between Tamis 3 and the Baird-Parker Agar plate count. The total results of microbiological tests showed that there were three correlations of the counts in selective culture media. Fifty-two possibilities were analysed and there was a negative correlation between no-bacterial-growth mastitis rates and log10 of KF Streptoccocus Agar plate count and there were two positive correlations between coagulase-positive staphylococci and log10 of Baird-Parker Agar plate count and Blood Agar plus potassium tellurite plate count.

Impact of milk protein composition on casein (CN) retention in curd during the milk coagulation process was studied using a model cheese making system. Individual milk samples from 110 cows in mid lactation of the Swedish Red and Swedish Holstein breeds with known genotypes of β-casein, κ-casein and β-lactoglobulin were defatted, coagulated with chymosin, subjected to syneresis and subsequent pressing simulated by centrifugation. The results indicated that κ-casein concentration of milk plays an important role in the curd formation process and initial syneresis (whey after cutting), whereas an increased CN ratio was associated with less casein in whey after simulated pressing. Increased κ-casein concentration of milk also characterized the milk samples with no measurable loss of casein in whey, compared with milk samples with casein lost in whey, both after cutting and after simulated pressing. Concentrations of αs1-casein, β-casein, and total casein in milk were positively associated with fresh curd yield, which showed a strong correlation with amount of casein retained in curd. No effect of protein genotype on fresh curd yield or casein in whey was found. The β-lactoglobulin BB genotype was associated with increased casein retention in curd, most likely due to the association of this genotype with CN ratio.

Social rank in cervids and other mammals is not entirely predicted by body weight, but in most cases influences access to food directly. Milk provisioning depends on maternal weight and on daily food intake. Usually, body weight, body condition, age and social rank are inter-correlated making it very difficult to discern the relative importance of each variable to milk production. This study used path analysis to assess direct versus indirect effects of these variables on milk production of 62 Iberian red deer hinds (Cervus elaphus hispanicus). Once the known direct effects of body weight and body condition were set as fixed, hind age and social rank did not affect milk production directly. In contrast, they exerted an indirect influence through the correlation both with hind body weight and body condition. Body weight exerted an effect on milk production nearly twice as great as that of body condition. This study shows, for the first time in a wild mammal, the relative importance of social rank, body weight, body condition and age in affecting milk production ability.

The adaptation of a colorimetric technique for the analysis of cholesterol in raw milk is presented. Performance quality was satisfying (mean intra-assay coefficient of variation (CV) 4·8, inter-assay CV 9·1%, linearity between 0 and 7 mm, recovery of spiked cholesterol into raw milk 98·1 and 106·3%). However, the milk fat extraction must be carried out within the 48 hours following milk sampling. When performing sampling, the significant variation of milk cholesterol composition during the milking process has to be taken into account.

This field study focused on the possible effects of increased milking frequency (milking four times a day in comparison with milking twice a day) on clinical and bacteriological cure rates of clinical, antibiotically treated mastitis cases. Parameters tested were clinical, microbiological and full (cytomicrobiological) cure as well as the development of milk yield after the clinical mastitis episode. Cows from a large dairy herd meeting the study criteria (n=93) were assigned to two treatment groups by a systematic randomization scheme (blocked by body temperature <=or >39·5°C). Both groups were randomly divided by experimental treatments: a) antibiotic intramammary treatment and milking 2-times a day; b) antibiotic intramammary treatment and milking 4-times a day. Treatments were initiated before the culture results were known. Cows were surveyed and evaluated on days 1–6, 24 and 31. No significant differences between treatment and control groups regarding clinical cure, microbiological cure, full cure and milk production could be established. Applying a 4-times a day milking regime did not lead to any significant effect, either positive or negative. Therefore, the results suggest that milking 4-times a day as a supporting therapy for mild, moderate and severe antimicrobially treated mastitis cases cannot be recommended.

Two body condition scoring systems were compared for assessing body condition of cows at the Scottish Agricultural College's Crichton Royal Farm. The weekly body condition scores (BCS) were collected for a period of 12 weeks (5 September–21 November). Scores were obtained using the primary systems utilized within the UK and USA. The USBCS were obtained by the same evaluator each week, while the UKBCS were obtained by two different evaluators alternating between weeks. Paired scores (n=2088) between the two systems within week were moderately correlated (r=0·75, P<0·0001). Regression equations to convert scores between the two systems were created using the GLM procedure of SAS (SAS Institute Inc., Cary NC, USA). The simple GLM models to convert from UK to US scores and US to UK scores were USBCS=1·182+0·816 * UKBCS (R2=0·56) and UKBCS=0·131+0·681 (R2=0·56), respectively. These equations may be used to interpret scores within the literature obtained using these two BCS systems, although they must be used with caution.

The dry period plays an important role in maintenance of udder health. Cows are most susceptible to intramammary infections (IMI) after dry-off and near parturition and drying-off procedures may affect the likelihood of IMI at calving. The objective of this study was to evaluate the association of milk yield and infection status at dry-off with the likelihood of IMI at calving by examining different drying-off methods. Cows (n=112) at the Ohio State University Waterman Dairy Teaching and Research Herd were randomly assigned to either an intermittent or a standard, twice-daily milking group 1 week prior to dry-off. All quarters of all cows in the herd were treated with an antibiotic dry-cow product after the last milking. Milk samples were collected 1 week prior to dry-off (pre-dry), on the day of dry-off, and within 3 d of parturition to determine infection status of the quarters. Association between IMI at calving and cumulative milk yield for the final week of lactation and drying-off method was examined using generalized estimation equations with logic link, accounting for potential confounders, such as pre-dry and dry-off infection status, and for the correlated data structure due to quarters clustered within cows. Intermittent milking significantly reduced milk yield at the end of lactation. Increasing cumulative milk yield during the last week of lactation was significantly associated with a greater probability of IMI at calving for quarters that were uninfected prior to dry-off: uninfected quarters of cows producing more than 115 kg during the last week of lactation were 7·1-times more likely to be infected at calving (P=0·0081) than uninfected quarters of cows producing less than 75 kg. Even though the overall cure rate over the dry period was relatively high at 84%, the odds of a quarter being infected at calving was 7·6- and 3·3-times higher if it was infected at dry-off with major pathogens (P<0·0001) or minor pathogens (P=0·028), respectively, compared with an uninfected quarter at dry-off. The results suggest that decreasing milk yield prior to dry-off may serve as an effective means to maintain good udder health in a herd.

The objective was to investigate the changes occurring in the activities of the enzymes lactate dehydrogenase (LDH), alkaline phosphatase (ALP) and aspartate aminotransferase (AST) in sheep and goat milk as a result of subclinical intramammary infections (IMI) and to evaluate the use of these enzymes for the diagnosis of subclinical IMI in dairy sheep and goats. A total of 206 samples of sheep milk and 162 samples of goat milk, obtained from equal udder halves, were used in the study. For each species they were divided into two groups: a no-infection group and a subclinical infection group. Activities of LDH, ALP and AST were significantly higher in the subclinical infection group than in the no-infection group (P<0·05) in both sheep (LDH: 350·42±11·25 v. 120·91±4·41; ALP: 2773·43±105·18 v. 2189±94·24; AST: 29·57±0·74 v. 17·32±0·46) and goats (LDH: 354·07±13·33 v. 103·79±3·75; ALP: 311·13±25·74 v. 137·24±19·62; AST: 27·59±6·42 v. 15·87±0·45). The activity of LDH was identified as indicator for subclinical IMI in both sheep and goats. The optimum cut-off values for LDH activity, offering the highest diagnostic sensitivity (DSn) and diagnostic specificity (DSp), determined by receiver operating characteristic (ROC) analysis, were at 197 U/l, 185 U/l and 197 U/l for sheep, goats and both species, respectively. DSn for sheep, goats and both species at these cut-off values was 92·8%, 98·2% and 94·0%, whereas DSp was 95·4%, 96·3% and 96·3%, respectively. It was concluded that the determination of LDH activity in milk serum is a sensitive and reliable method for the detection of subclinical IMI in dairy sheep and goats.

Conventional methods for the identification of Listeria in foodstuffs are generally cumbersome and time consuming. The use of primary enrichment in half strength Fraser broth and the use of PALCAM agar were assessed in comparison with API Listeria and polymerase chain reaction (PCR) for their ability to accurately detect and confirm the presence of List. monocytogenes in milk products. The aim of our work was to detect List. monocytogenes in domestic unpasteurised milk, fresh cheese and cream of raw milk taken from four different district of Zagreb-Croatia using conventional (microbiological and biochemical – API test) and PCR methods. Of the 180 milk products samples tested, 27·6% were presumptively positive for Listeria on PALCAM agar. Only 21·3% of samples were confirmed to be positive for Listeria by API Listeria test, and 17·3% were confirmed to be positive for List. monocytogenes by PCR amplification of the hly gene (64 bp). PCR was able to eliminate the false positive and detect all List. monocytogenes in the milk products, unlike the conventional methods used in the industry. These results indicate a low presence of this pathogen in this area (Zagreb) of Croatia. PCR proves to be a sensitive and rapid technique to be included in the procedure of detection of List. monocytogenes in food products and this method is considerably faster than current standard methods.

The aim of the study was to evaluate the profile of antioxidant parameters in ewes' colostrum and milk in relation to breed during 5 d post partum. Total antioxidant capacity (TAC) was analysed and the activity of the enzymic antioxidants, glutathione peroxidase (GSH-Px) and glutathione transferase (GSH-Tr), as well as the concentration of the non-enzymic antioxidants, vitamin C, vitamin A and β-carotene, were measured. Samples were collected from healthy animals belonging to two ewe breeds: Berrichon du Cher (n=15) and Uhrusk (n=15) kept in the Podlasie Province (Poland). Colostrum was sampled directly after parturition, as well as after 12, 24 and 48 h later and milk was sampled 5 d after parturition. Colostrum and milk for the evaluation of all parameters except for vitamin A and β-carotene were centrifuged, and the supernatant was used for further analysis. Spectrophotometric methods were used for biochemical measurements. The results showed dynamic changes of antioxidative parameters within the time period examined. TAC values and GSH-Px activity increased significantly during the experiment. GSH-Tr activity showed a similar tendency in Uhrusk ewes but an opposite relationship in Berrichon du Cher. Concentrations of examined vitamins followed the increasing trends noticed in the activities of antioxidative enzymes. Moreover, differences between breeds in the evaluated parameters were detected; these differences were not unequivocal however. The results are also a source of not previously published physiological antioxidant profile in colostrum and milk of ewes over the post-partum period.