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.

During the normal course of lactation, mammary metabolic activity and blood flow are closely correlated. Six lactating goats were used in this experiment to test the hypothesis that the capillary network and the capillary enzyme, carbonic anhydrase (CA; EC 4.2.1.1) are important regulatory factors involved in the coordination of mammary blood flow (MBF) and metabolic activity. Milk vein blood velocity was determined as a measure of MBF, and fine needle mammary biopsies were obtained at different time points during lactation and by the end of a 14-d bovine somatotropin (BST) treatment initiated 3 months post partum. In mammary sections, CA activity was determined histochemically and alveolar and capillary structures by image analyses upon azure blue staining. In early lactation, alveoli were large and surrounded by many small capillaries with high CA activity. As lactation progressed, capillaries almost tripled in size, whereas number of capillaries surrounding each alveolus decreased by 1/3, and CA activity more than halved. BST treatment did not affect capillary traits but increased number of alveoli in mammary sections, and BST thus appeared to be targeted mostly towards the mammary epithelial cell. Milk vein blood velocity decreased over the course of lactation, when capillary area markedly increased, suggesting that control of mammary blood perfusion is not at the level of the capillary itself, but at pre- or post-capillary sites. We suggest that the observed changes in capillary diameter and CA activity with progressing lactation contributes to reduce efficiency of nutrient and waste product exchange across the capillary-mammary epithelial cell barrier, and this could be an important factor in regulation of mammary (epithelial cell) metabolic activity and lactation performance.

The aim of this study was to evaluate the milk quality of Massese ewes receiving diets with different forage:concentrate ratios (FC ratio), specially on milk fat globules characteristics and fatty acids composition. The diet is one of the main environmental factors that influence the lipidic content of milk. A trial was carried out on twenty ewes, which had been subdivided into two homogeneous groups and kept indoors at 25 days post partum. The experiment lasted 60 days, from 40 to 100 days post partum and the animals were fed two diets that differed in terms of the FC ratio: 60:40 and 40:60, as fed. The results obtained in this study showed that a greater proportion of forage, compared with an higher percentage of concentrate, led to an increase in the percentage of fat (+8·66%) and to a decrease in the percentage of milk fat globules with a size between 2 and 5 μm (−17·32%). However, the average diameter was not affected. There was also a decrease in the percentages of some medium chain fatty acids (C12:0, C14:0; −14·89% and −4·03 respectively) and an increase in mono and polyunsaturated ones such as trans11-C18:1 (+31·71%), total CLA (+22%), EPA (+18·18%) and DHA (+66·67%). In conclusion, a greater proportion of forage seem to improve the milk fatty acid profile by the increase of some fatty acid identified has being beneficial for human health.

Our previous studies demonstrated prompt elevation of proteinase activity in mammary secretion of drying-off cows and goats. The current study examined the progressive changes in composition of cow mammary secretion following drying-off and, in parallel, characterized the mode of peptide neogenesis using matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS) and liquid chromatography-electrospray-ionization (LC-ESI) MS/MS. The results show that the percentage of casein of total milk protein at time of drying-off was 76%, which dropped to 41%, 24%, and 16%, respectively, 1, 2, and 3 weeks after drying-off. Levels of β-lactoglobulin and α-lactoalbumin in mammary secretions of drying-off cows decreased prominently while levels of lactoferrin, BSA, and casein derived-proteins increased concomitantly compared with regular milk. A fractionation procedure was applied to remove molecules larger than 10 kDa before MALDI-TOF MS and LC-ESI MS/MS and the results show that the MALDI-TOF MS peptide profile of mammary secretion ranging from m/z 600 to 4000 was apparently modified after drying-off for 1 week, whereas species 1590 m/z and 2460 m/z were most obviously enriched compared with regular milk. LC-ESI MS/MS results were used to map peptide sequence with Mascot search server and under no post translational modification to reduce database size and 202 novel β-casein-derived peptides were successfully identified in mammary secretion after drying-off for 1 week in contrast to regular milk. Accordingly at least 48 additional cleavage positions were assigned on β-casein for mammary secretion. Among the 202 novel peptides, 5 are homologous with confirmed opioid agonists, angiotensin 1-converting enzyme inhibitors, or immuno-modulators. In conclusion, peptides are released in situ from milk proteins within short intervals following drying-off in cows. They might play roles in the transition of mammary glands from lactating to non-lactating. With specified post-translational modifications and focused functional screening, novel peptides are yet to be discovered in dry cow mammary secretion.

A study was carried out to evaluate how the energy level of the diet can affect milk production and quality in Girgentana lactating goats in relation to polymorphism at the αs1-casein (CSN1S1) genotype locus. Twenty-seven goats, homogeneous for milk production (1·5±0·3 kg/d), days of lactation (90±10 d) and body weight (35·8±5·5 kg) were selected on the basis of their CSN1S1 genotype, as follows: nine goats homozygous for strong (AA) alleles, nine goats homozygous for weak alleles (FF) and nine goats heterozygous (AF). The goats were used in a 3×3 factorial arrangement of treatments, with three genotypes (AA, FF, AF) and three diets at different energy levels (100%, 65% and 30% of hay inclusion). The experiment consisted of three simultaneous 3×3 Latin squares for the three genotypes, with one square for each level of hay inclusion in the diet. All the animals were housed in individual pens. Each experimental period lasted 23 d and consisted of 15 d for adaptation and 8 d for data and sample collection, during which the goats received the scheduled diet ad libitum. The animals were fed three different diets designed to have the same crude protein content (about 15%) but different energy levels: a pelleted alfalfa hay (H100) and two feeds including 65% (H65) and 30% (H30) of alfalfa hay (respectively 1099, 1386 and 1590 kcal NE for lactation/kg DM). All the diets were ground and pelleted (6 mm diameter). AA goats were more productive than AF and FF goats (respectively: 1419 v. 1145 and 1014 g/d; P=0·002). Indeed the interaction energy level×genotype was significant (P=0·018): in fact AA goats showed their milk increase only when fed with concentrates. Differences in protein and in casein levels between the three genotypes were in line with results expected from the different allele contribution to αs1-casein synthesis. Milk urea levels were significantly lower in AA goats compared with AF and FF genotypes (respectively 32·7 v. 40·4 and 40·4 mg/dl; P=0·049) and significantly lower when goats were fed with 65H and 30H diets than with 100H diet (respectively 37·4 and 34·3 v. 41·7 mg/dl; P<0·001). Indeed, a significant interaction genotype×diet (P=0·043) occurred for milk urea, which was significantly lower in AA goats but only when fed with concentrates (65H and 30H). Blood concentrations of energy indicators (glucose, non-esterified fatty acids and beta-hydroxybutyric acid) were not influenced by genotype. The results confirm that strong alleles are associated with a greater efficiency of feed utilization and seem to show that a high energy level of the diet can further improve this efficiency.

The objective of this study was to investigate the effect of ruminal infusion of soybean oil (SBO) with either a moderate- or high-forage diet on fat concentration, yield and composition in milk from dairy cows. Six rumen-fistulated Holstein dairy cows (639±51 kg body weight, 140±59 days in milk) were used in the study. Cows were randomly assigned to one of two dietary treatments, a high forage:concentrate (HFC, 74:26) or a moderate forage:concentrate (MFC, 56:44) total mixed ration. Cows were fed at 08.00 and 13.00 h and pulse-dosed ruminally at 13.00 h over a 10-min duration with 2% of diet dry matter of SBO. Ruminal pH was recorded continuously. Cows receiving the MFC treatment had lower daily mean ruminal pH and ruminal pH was below 6·0 for a longer duration compared with the HFC treatment (640 vs. 262 min/d, P<0·05). Cows receiving the MFC treatment had a greater reduction (diet by week interaction, P<0·05) in milk fat concentration and yield than cows receiving the HFC treatment (42 vs. 22% and 45 vs. 21%, respectively). Additionally, cows receiving the MFC diet had a greater reduction in milk fat concentration (g/100 g FA) of FA <C16 (14 vs. 8%), and a greater increase in concentration of FA >C16 (17 vs. 9%), trans-10 18:1 (159 vs. 21%) and trans-9, cis-11 conjugated linoleic acid (121 vs. 55%) (P<0·05) compared with cows receiving the HFC diet. This study demonstrated that cows fed the MFC diet had lower ruminal pH and showed a greater rate of milk fat depression when infused with SBO.

The potential of ethidium monoazide (EMA) real-time PCR method based on molecular beacon probe for rapid detection of viable bifidobacteria present in probiotic yogurt was evaluated in this work. A real-time PCR with molecular beacon assay was developed to determine genus Bifidobacterium quantitatively in order to increase the sensitivity and specificity of assay. EMA was used to treat probiotic yogurt prior to DNA extraction and real-time PCR detection to allow detection of only viable bacteria. The primer set of Bif-F/Bif-R which is genus-specific for Bifid. was designed. The specificity of the probes ensures that no signal is generated by non-target amplicons. Linear regression analysis demonstrated a good correlation (R2=0·9948) between the EMA real-time PCR results and the plate counting, and real-time quantitative PCR results correlated adequately with enumeration of bifidobacteria by culture for commercial probiotic yogurt. This culture-independent approach is promising for the direct and rapid detection of viable bifidobacteria in commercial probiotic yogurt, and the detection can be carried out within 4 h. The detection limit for this method is about 104 cell/ml. In conclusion, the direct quantitative EMA real-time PCR assay based on molecular beacon described in this research is a rapid and quantitative method.

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.

In order to contribute to the breeding programmes of Asian water buffalo, the aim of this study was to analyse the influence of genetic effects in the stayability of Murrah dairy buffaloes. The stayability trait (ST) was defined as the female's ability to stay in the herd for one (ST1), two (ST2), three (ST3), four (ST4), five (ST5) or six years (ST6) after the first calving. The same trait was also considered as continuous and was designated stayability in days up to one (STD1), two (STD2), three (STD3), four (STD4), five (STD5) or six years (STD6) after the first calving. Data from 1016 females reared in nine herds located in the State of São Paulo, Brazil, were analysed. Statistical models included the additive genetic effect of the animal and the fixed effects of the buffalo breeding herd, birth year and birth season. Additive effects for ST were estimated by approximate restricted maximum likelihood using a threshold model, while for STD, the additive effects were estimated by restricted maximum likelihood. Heritability estimates were lower for ST, except for ST1, (0·11±0·07, 0·17±0·06, 0·23±0·06, 0·16±0·08, 0·14±0·09 and 0·16±0·10 for ST1, ST2, ST3, ST4, ST5 and ST6, respectively) when compared with STD (0·05±0·06, 0·18±0·08, 0·40±0·10, 0·49±0·11, 0·41±0·11 and 0·30±0·13, for STD1, STD2, STD3, STD4, STD5 and STD6, respectively). Considering the values of heritability and owing to the serial nature of STD to a specific age, selection for STD3 should have a favourable influence on STD to other ages.

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.

The objective was to evaluate the effects of an inhibitor of α-amylase and glucosidase (acarbose, Pfizer Limited, Corby, UK) on ruminal fermentation, blood metabolism and microbial profile in dairy cows in a 2×2 cross-over experiment. Eight Holstein cows fitted with rumen cannulas (milk yield, 24·3±2·35 kg/d, body weight, 622±54 kg, days in milk, 183±67, 5 multiparous and 3 primiparous) were used. Treatments were: control (no additive, CTR) and α-amylase and glucosidase inhibitor (0·75 g acarbose-premix/cow per d, AMI). Animals were given ad-libitum access to a high non-fibre carbohydrate (NFC) partial mixed ration (PMR) containing 17·6% crude protein, 28·3% neutral detergent fibre, and 46·5% NFC in the dry matter and supplementary concentrate during milking. Blood samples were taken to determine blood glucose, insulin and urea within the first hour after the morning feeding on two separate days in each period. Samples of ruminal contents were collected during 3 d in each period at 0, 4 and 8 h after feeding to determine volatile fatty acid and ammonia-N concentrations and to quantify protozoa, Streptococcus bovis and Megasphaera elsdenii. Rumen pH was recorded electronically at 22-min intervals during 6 d in each period. Results were analysed using a mixed-effects model. Cows on AMI treatment spent less time with ruminal pH <5·6 compared with cows in the CTR group (3·74 and 6·52±0·704 h/d, respectively). Cows in the AMI group had greater daily average pH compared with those in the CTR group (6·05 and 5·92±0·042, respectively). AMI animals tended (P=0·09) to have lower Str. bovis to Meg. elsdenii ratio than CTR (4·09 and 26·8±12·0, respectively). These results indicate that dietary supplementation with acarbose in dairy cattle fed high-production rations may be effective in reducing the time for which rumen pH is suboptimal, with no negative effects on ruminal fermentation and blood metabolites.