In this Research Paper we hypothesised that the temporary insulin resistance seen during the transition period in dairy cows may cause significant differences in the efficacy of PG at different sampling periods and that in some cases this effect will be dose dependent. Eighty four sampling sets were generated by studying 7 multiparous Holstein cows repeatedly at 4 sampling periods of 3 d length (dry cows: days 40, 39 and 38 antepartum; close up cows: days 10, 9 and 8 antepartum; fresh cows: days 3, 4 and 5 post-partum; lactating cows: days 38, 39 and 40 post-partum). On each of these days 3 h after morning feeding propylene glycol was drenched in different dosages of 100, 300 or 500 ml once per day (cross over study). The different doses were applied in an alternating order (Latin square). Blood samples were taken before, every 30 min up to 4 h, after 6 and 12 h after PG application. Following parameters have been measured: insulin, non-esterified fatty acids (NEFA), betahydroxybutyrate (BHB), bilirubin, cholesterol, potassium, aspartate aminotransferase (AST) and glutamate dehydrogenase (GLDH). Revised Quantitative Insulin Sensitivity Check Index (RQUICKI) was calculated. It was found that glucose, insulin, NEFA, BHB, bilirubin and potassium concentrations were influenced differently by the three defined dosages of propylene glycol at four different sampling periods. Whereas RQUICKI, cholesterol, AST and GLDH did not differ between the sampling periods and treatments. The major results of the study are that the effect of PG is dose-dependent and that the effect of PG is depending on the time of application according to calving. It can be concluded that in fresh cows higher dosages are necessary to provoke similar effects in comparison to dry, close up and lactating cows. Although the study did not compare to topdressing of PG from the results it is reasonable to believe that bolus application of a specific PG volume is necessary to provoke the effect.

We examined the hypothesis that rumination time (RT) could serve as a useful predictor of various common diseases of high producing dairy cows and hence improve herd management and animal wellbeing. We measured the changes in rumination time (RT) in the days before the recording of diseases (specifically: mastitis, reproductive system diseases, locomotor system issues, and gastroenteric diseases). We built predictive models to assess the association between RT and these diseases, using the former as the outcome variable, and to study the effects of the latter on the former. The average Pseudo-R2 of the fitted models was moderate to low, and this could be due to the fact that RT is influenced by other additional factors which have a greater effect than the predictors used here. Although remaining in a moderate-to-low range, the average Pseudo-R2 of the models regarding locomotion issues and gastroenteric diseases was higher than the others, suggesting the greater effect of these diseases on RT. The results are encouraging, but further work is needed if these models are to become useful predictors.

This research communication validates an algorithm to monitor natural occurrence of locomotor behaviours in dairy Mediterranean buffalo based on the output of a 3-dimensional accelerometer (RumiWatch®, pedometer). Several characteristics of the locomotor behaviour were detected with a very high (up-right, lying and standing time) or high degree of correlation (walking time and number of strides) and a low mean difference with the video recording. The proportion of correctly detected events exceeded 99 % for the following variables: stand up and lie down events, as well as number of lying, standing or walking bouts. The mean relative measurement error was less than 10 % for the variables: lying, standing, up-right times and number of strides as compared with gold standard. This new algorithm may represent the base for a future early and real-time disease warning system aiming to gain higher health standard in these ruminants.

Interbeat interval data were collected from 219 Holstein cows in 2 smaller-scale farms and 3 larger-scale farms to investigate the effects of posture (standing vs. lying), rumination (rumination vs. no rumination) and feeding on baseline values of heart rate (HR) and heart rate variability (HRV) parameters reflecting vagal and sympathetic activity. A General Linear Model was used for detecting factors (parity, milking technology, herd size) having possible effects on HRV calculated for undisturbed lying posture. Basal values of cardiac parameters were also compared between larger and smaller-scale farms. Neither parity nor milking technology affected HRV parameters. Sympathetic activity increased in the order of lying, ruminating when lying, standing, ruminating when standing and feeding on both sizes of farms. Vagal activity decreased in that order in both lower- and larger-scale farms. Rumination caused an increase in HR and a decrease in vagal tone in both lower- and larger-scale farms and an increase in sympathetic activity during lying in both farm sizes. Basal vagal activity was lower in larger-scale farms compared to smaller-scale farms, while greater sympathetic activity was found in cows housed on larger-scale farms. Our findings demonstrate that reference values of HRV parameters in lactating dairy cows cannot be generally defined for Holstein cattle as they are affected by physical activity and herd size. Higher HR and sympathetic activity at rest in larger-scale farms compared to farms with lower cow population might be associated with higher levels of social stress and therefore should be considered as a potential welfare concern.

The purposes of this study were to investigate the systemic and characteristic metabolites in the serum of dairy goats induced by aflatoxin B1 (AFB1) exposure and to further understand the endogenous metabolic alterations induced by it. A nuclear magnetic resonance (NMR)-based metabonomic approach was used to analyse the metabolic alterations in dairy goats that were induced by low doses of AFB1 (50 µg/kg DM). We found that AFB1 exposure caused significant elevations of glucose, citrate, acetate, acetoacetate, betaine, and glycine yet caused reductions of lactate, ketone bodies (acetate, β-hydroxybutyrate), amino acids (citrulline, leucine/isoleucine, valine, creatine) and cell membrane structures (choline, lipoprotein, N-acetyl glycoproteins) in the serum. These data indicated that AFB1 caused endogenous metabolic changes in various metabolic pathways, including cell membrane-associated metabolism, the tricarboxylic acid cycle, glycolysis, lipids, and amino acid metabolism. These findings provide both a comprehensive insight into the metabolic aspects of AFB1-induced adverse effects on dairy goats and a method for monitoring dairy animals exposed to low doses of AFB1.

High concentrations of non-esterified fatty acid (NEFA) and β-hydroxybutyrate (BHBA) in cows’ blood caused by ketosis are associated with inflammatory states. We hypothesised that ketosis in postparturient dairy cows would result in altered levels on inflammation-related proteins not only in plasma but also in the milk fat globule membranes (MFGM). Thirty cows were selected from a dairy farm in Heilongjiang, China. Inflammatory milk fat globule membrane proteins were detected using ELISA kits, and a fully automatic biochemical analyser was used to measure the concentrations of BHBA, NEFA, glucose (GLU) and triglyceride (TG) in plasma. MFGM protein from milk of ketotic cows contained significantly different concentrations of acute-phase response proteins (complement C3 (C3), prothrombin (F2), alpha-1-acid glycoprotein (ORM1), inter-alpha-trypsin inhibitor heavy chain H4 (ITIH4), alpha-2-HS-glycoprotein (AHSG), complement C9 (C9), complement regulatory protein variant 4 (CD46)) in comparison with milk from non-ketotic cows. Blood concentrations of C3, complement C9 (C9), tumour necrosis factor α (TNFα), MFGM C3, monocyte differentiation antigen CD14 (CD14) and ORM1 levels were correlated with energy balance. ITIH4 and CD46 increased, and AHSG and ORM1 decreased before the onset of ketosis. These biomarkers offer potential as predictors and monitors of ketosis in at-risk cows.

The work reported in this Research Communication describes the modification in epithelial cell populations during the first and the last month of milking in Holstein Friesian cows that have undergone different management during the dry period, and we report the differential expression of CD49f+ and cytokeratin18+ cell subpopulations. Twenty six cows were randomly divided into 2 balanced groups that were housed at stocking density of either 11 m2 (CTR) or 5 m2 from 21 ± 3 d before the expected calving until calving. Cells collected from milk samples taken in early lactation and late lactation were directly analysed for CD45, CD49f, cytokeratin 14, cytokeratin 18 and cell viability. We observed a differential expression with a significant reduction in CD49f+ (P < 0·01) and cytokeratin 18+ (P < 0·05) cells in early lactation. Differences were still evident in late lactation but were not significant. These observations suggest that mammary epithelial cell immunophenotypes could be associated with different animal management in the dry period and we hypothesise they may have a role as biomarkers for mammary gland function in dairy cows.

Bovine mastitis, an inflammation of the udder, results in reduced milk production and poor milk quality. Mastitis is usually, but not always, a response to pathogen infection. High milk yield can produce oxidative stress in the mammary tissue. High milk yield is also known to be associated with bovine mastitis. Thus, in the current study, we hypothesised that oxidative stress increases inflammatory responses in bovine mammary cells. To examine the hypothesis, we produced cellular oxidative stress and investigated resulting inflammatory responses in bovine mammary alveolar cells (MAC-T). To produce oxidative stress, cells were treated with the reactive oxygen species (ROS; e.g., superoxide anion)-producing agent, menadione (MD; 0–10 µm; 6 h). To ensure the ROS-induced responses, cells were pretreated with an antioxidant NAC (0–10 mm; 1 h). Results showed that MD elevated intracellular ROS levels and protein expression of cyclooxygenase-2 (COX-2), a biomarker of inflammation. Pretreatment of cells with NAC attenuated MD-induced COX-2 expression by scavenging intracellular ROS and enhancing intracellular glutathione levels. MD-induced COX-2 expression was mediated by activation of extracellular signal receptor-activated kinase 1/2 (ERK1/2), Akt, and nuclear factor-kappa B (NF-κB). NAC attenuated activation of these intracellular signalling molecules. Treatment of cells with pharmacological inhibitors for ERK1/2, Akt, and NF-κB confirmed the association of these signalling pathways in MD-induced COX-2 expression. These results support our hypothesis that oxidative stress, which is found in high-yielding dairy cows, can produce cellular inflammation in bovine mammary alveolar cells and prevention of oxidative stress can attenuate such pathological responses. This may be relevant for cases of clinical mastitis for which no pathogen can be isolated.

The objective of this Research Communication was to use polymerase chain reaction-single stranded conformational polymorphism (PCR-SSCP) analysis to investigate a region of the bovine TLR4 gene (TLR4) in pasture-fed New Zealand (NZ) Holstein-Friesian × Jersey (HF × J) cross dairy cows and to determine whether gene variation was associated with milk production traits. Genetic variation was observed, with two variants (A and B) containing a single nucleotide polymorphism (SNP) (c.2021C/T) that was non-synonymous and putatively results in a p.Thr674Ile substitution in the transmembrane/cytoplasmic domain of TLR4. Variant A was associated with higher milk yields, but lower milk fat percentages, whereas B was associated with lower milk yields, but higher fat and protein percentages. Cows of genotype AA produced more milk than AB or BB cows, but the milk produced by AA cows contained less fat than AB or BB cows.

This Research Communication describes the association between genetic variation within the prolactin (PRL) gene and the milk production traits of Italian Mediterranean river buffalo (Bufala mediterranea Italiana). High resolution melting (HRM) techniques were developed for genotyping 465 buffaloes. The association of genetic polymorphism with milk production traits was performed and subsequently the effects of parity and calving season were evaluated. Single nucleotide polymorphisms (SNPs) were identified at exons 2 and 5 and at introns 1 and 2. All the SNPs were in Hardy–Weinberg equilibrium, and statistical analysis showed that the polymorphism of intron1 was significantly (P < 0·05) associated with milk yield, milk protein content and peak milk yield. The average contribution of the intron1 genotype (r2intron1) to total phenotypic variance in milk production traits was 0·09, and the TT genotype showed lower values than CC and CT genotypes. A nonsynonymous SNP was identified in exon 2, which resulted in an amino acid change from arginine to cysteine. Moreover, the polymorphism of exon 2 was associated significantly with milk fat content (P < 0·05), and the buffaloes with TT genotype showed higher total fat content than the buffaloes with CT genotype. These findings provide evidence that polymorphisms of the buffalo PRL gene are associated with milk production traits and PRL can be used as a candidate gene for marker-assisted selection in Italian Mediterranean river buffalo breeding.

The objectives of this study were to estimate the genetic parameters and genetic trends for different measures of persistency (PM) for milk yield in Iranian buffaloes using a random regression test day model. Test day records of the first three lactations of buffaloes within 715 herds comprising 43 014, 38 941 and 33 937 records, respectively, were obtained from the Animal Breeding Center of Iran during 1992 to 2012. Legendre polynomial functions were chosen to fit the lactation curves in the framework of a random regression test day model for estimating (co)variance components. Three measures of persistency were modified based on the lactation curve conditions of buffaloes: (1) The average of estimated breeding values (EBVs) for test day milk yields from day 226 to day 270 as a deviation from the average of EBVs from day 44 to day 62 (PM1), (2) A summation of contribution for each day from day 53 to day 247 as a deviation from day 248 (PM2), (3) The difference between EBVs for day 257 and day 80 (PM3). The highest heritability estimates were observed for PM2 over the first three lactations (0·29, 0·80 and 0·42, respectively). Estimates of genetic correlations between persistency measures and milk yield varied from −0·48 to 0·29 (first lactation), from −0·12 to 0·48 (second lactation), and from −0·62 to 0·63 (third lactation). Estimate of genetic trend for first lactation PM2 was negative (−0·0088 ± 0·0043; P < 0·05). It is essential to include milk yield persistency in the breeding goal of Iranian buffaloes along with other important traits such as milk yield or reproductive traits.

In this Research Communication we assessed factors affecting colostrum quality of dairy Lacaune ewes using the Brix-refractometer. Colostrum from 536 lambings from one commercial intensive dairy Lacaune farm were analysed for the following factors with potential influence in colostrum quality: (1) ewe parity (n = 84–132), (2) length of previous dry period (PDP) (n = 23–214), (3) age at first lambing (AFL) of primiparous ewes (n = 9–88), (4) lambing season (n = 192 or 344), and (5) year (2011–2013, n = 142–203). Parity significantly affected colostrum quality, with primiparous ewes showing the highest Brix refractometer values (22·6 ± 5·6%, P < 0·0001), though values were similar among multiparous ewes. PDP length also significantly affected colostrum quality: ewes with the shortest PDP showed the worst quality (16·8 ± 4·2%, P < 0·0001), with quality gradually rising with PDP length. Colostrum quality was significantly higher in 2011 (21·0 ± 5·2%) than in 2012 or 2013 (P < 0·0001); this likely reflects the several-fold greater proportion of animals with long PDP in 2011. In contrast, neither AFL nor lambing season significantly affected colostrum quality. These results suggest that parity and PDP length can substantially affect ovine colostrum quality of dairy ewes under intensive management conditions and they further show the usefulness of the Brix refractometer for providing a rough estimation of colostrum quality on-farm. However, further studies are needed to determine a validated cut-off Brix value for identifying good-quality colostra in ovine species.

The study investigated the seasonal changes in the compositional, physicochemical and processing characteristics of milk from a mixed-herd of spring- and autumn-calving cows during the year 2014–2015. The volume proportion of autumn-calving milk (% of total milk) varied with season, from ~10–20 in Spring (March–May), 5–13 in Summer (June–August), 20–40 in Autumn (September–November) and 50–100 in Winter (December–February). While all characteristics varied somewhat from month to month, variation was inconsistent, showing no significant trend with progression of time (year). Consequently, season did not significantly affect many parameters including concentrations of total protein, casein, whey protein, NPN, total calcium, pH, rennet gelation properties or heat stability characteristics. However, season had a significant effect on the concentrations of total P and serum P, levels of αs1- and β-caseins as proportions of total casein, casein micelle size, zeta potential and ethanol stability. The absence of a significant effect of season for most compositional parameters, rennet gelation and heat-stability characteristics suggest that milk from a mixed-herd of spring- and autumn-calving cows is suitable for the manufacture of cheese and milk powder on a year-round basis, when the volume proportion of autumn milk, as a % of total, is similar to that of the current study.

It has been well established that milk yield is affected both by milking frequency and due to the removal of residual milk, but the influence of a combination of these factors is unclear. In this study, four mid-lactation cows were used in a 4 × 4 Latin square design to test the hypothesis that the effects of more frequent milking and residual milk removal on milk yield and composition are additive and alter milk fatty acid composition. Treatments comprised two or four times daily milking in combination with (or without) residual milk removal over a 96 h interval preceded by a 2 d pretreatment period and followed by a 8 d washout in each 14 d experimental period. Milk was sampled at each milking for the analysis of gross composition and SCC. Samples of available and residual milk collected on the last milking during each treatment period were collected and submitted for fatty acid composition analysis. Increases in milking frequency and residual milk removal alone or in combination had no effect on milk yield or on the secretion of lactose and protein in milk. However, residual milk removal during more frequent milking increased milk fat yield. Milking treatments had no major influence on the fatty acid composition of available milk, but resulted in rather small changes in the relative abundance of specific fatty acids, with no evidence that the additive effects of treatments were due to higher utilisation of preformed fatty acids relative to fatty acid synthesis de novo. For all treatments, fat composition of available and residual milk was rather similar indicating a highly uniform fatty acid composition of milk fat within the mammary gland.

The experiments reported in this research paper describe the effects of β-galactosidase enzyme dose and cheese whey amount, on the maximum concentration and yield of galacto-oligosaccahride (GOS) and reaction time. The experimental plan was based on central composite rotational design (CCRD) and modelled by response surface methodology (RSM). The results indicate that the proposed mathematical models could adequately describe the concentration and yield of GOS and the reaction time within the limits of the factors that are being investigated. The variance analysis shows high values of coefficients of determination (>0·97) while no significant lack of fit was evident. Hence, the models could be employed to select reaction conditions applied in the manufacture of products enriched in bioactive compounds with high value-added.

Modulating conditions during ultrafiltration of skim milk appears to be a feasible strategy to obtain milk protein concentrates (MPC) with tailored functionalities. Adjustment of pH and process temperature attenuated properties of casein micelle resulting in enhanced emulsification capacity. Additional pre-treatment options such as addition of calcium chelators can further impact on the functionality of MPC by modifying the calcium distribution and casein micelle integrity. The objective of the project was to establish effects of pre-treating skim milk with calcium chelators (EDTA or citrate) in concentrations between 10 to 30 mm prior to UF on the physical properties of the feed, corresponding retentates and dried MPC, including particle size, zeta potential and calcium distribution in skim milk and the corresponding retentates, as well as the physical functionalities such as solubility, heat stability and emulsifying properties. Addition of calcium chelators (EDTA or citrate), at levels 20–30 mm concentrations reduced casein micelle size as well as total, soluble and ionic calcium contents that resulted in MPC with enhanced solubility and heat stability. The emulsion capacity was, however, improved only with EDTA at 10 mm concentration. The enhanced functionality is attributed to the reduced particle size resulting from the removal of calcium from the retentate that could modify micellar casein to an extent sufficient to cause such improvements.

We studied the role played by temperature and rennet concentration in the coagulation process for cheese manufacture and the evaluation of their kinetics. We concluded that temperature is the main factor that determines the kinetics. The rennet concentration was unimportant probably due to the fast action of the enzyme chymosin. The Dynamic light scattering technique allowed measuring the aggregate's size and their formation kinetics. The volume fraction of solids was determined from viscosity measurements, showing profiles that are in agreement with the size profiles. The results indicate that the formation of the aggregates for rennet cheese is strongly dependent on temperature and rennet concentration. The results revealed that at 35·5 °C the volume fraction of solids has the maximum slope, indicating that at this temperature the curd is formed rapidly. The optimal temperature throughout the process was established. Second-order kinetics were obtained for the process. We observed a quadratic dependence between the rennet volume and the volume fraction of solids (curd), thereby indicating that the kinetics of the curd production should be of order two.

Contradictory statements about the effects of pH change on crystallisation behaviour of lactose exist in the literature. Considering the importance of addressing the processability issue of acid whey, a systematic study is required to establish lactose crystallisation behaviour in the presence of LA and Ca at concentrations present in real acid whey waste streams emphasising impact of pH. Structural modifications of lactose were evident at elevated, more neutral pH in the presence of 1% w/w LA and 0·12% w/w Ca. These structural changes led to changes in the anomeric equilibrium of lactose, which manipulated the water–lactose behaviour and increased the crystallinity. Therefore, altering pH to 6·5 may be the solution to proper industrial processing of acid whey, enhancing the ability of lactose to crystallise properly.

Nutritional interventions are now recommended as strategies to delay Alzheimer's disease (AD) progression. The present study evaluated the neuroprotective effect (anti-inflammation) of lactic acid bacteria (either Lactobacillus fermentum LAB9 or L. casei LABPC) fermented cow's milk (CM) against lipopolysaccharide (LPS)-activated microglial BV2 cells in vitro. The ability of CM-LAB in attenuating memory deficit in LPS-induced mice was also investigated. ICR mice were orally administered with CM-LAB for 28 d before induction of neuroinflammation by LPS. Learning and memory behaviour were assessed using the Morris Water Maze Test. Brain tissues were homogenised for measurement of acetylcholinesterase (AChE), antioxidative, lipid peroxidation (malondialdehyde (MDA)) and nitrosative stress (NO) parameters. Serum was collected for cytokine analysis. CM-LAB9 and CM-LABPC significantly (P < 0·05) decreased NO level but did not affect CD40 expression in vitro. CM-LAB attenuated LPS-induced memory deficit in mice. This was accompanied by significant (P < 0·05) increment of antioxidants (SOD, GSH, GPx) and reduction of MDA, AChE and also pro-inflammatory cytokines. Unfermented cow's milk (UCM) yielded greater cytokine lowering effect than CM-LAB. The present findings suggest that attenuation of LPS-induced neuroinflamation and memory deficit by CM-LAB could be mediated via anti-inflammation through inhibition of AChE and antioxidative activities.