The data presented in this research communication represent the first comprehensive report on the prevalence of udder pathogens and their resistance in Slovak dairy cattle breeds. A total of 633 milk samples from 42 herds were tested. The most frequently isolated pathogens were coagulase negative staphylococci CNS or non-aureus staphylococci (NAS) followed by Escherichia (E.) coli, Staphylococcus (S.) aureus, Streptococcus (Str.) uberis and Streptococcus (Str.) agalactiae. Generally, isolated pathogens showed common resistance to aminoglycosides (streptomycin, neomycin), which are the most commonly used antibiotics to treat mastitis in Slovakia.

This Research Communication describes the efficacy of etamsylate to reduce haemolactia in dairy cows. A dairy cow with haemolactia produces milk that is reddish or pinkish due to the presence of blood. Haemolactia causes economic loss because bloody milk is rejected by the industry and the consumers. A total of 58 dairy cows with haemolactia were included in the study and randomly divided into treated (n = 31) and control (n = 27) groups. Treatment consisted of three consecutive daily doses of etamsylate at 15 mg/kg, delivered intramuscularly. Milk production was recorded daily for 7 d, whether or not blood was detected in milk. The mean number of days with the presence of blood in milk in the treatment group was significantly lower (3·4 d) than in the control group (4·9 d). Treatment with etamsylate did not significantly affect milk yield. In conclusion, treatment with etamsylate reduces the number of days blood is observed in milk and it does not have any negative effect on milk production.

This research communication aims to test the hypothesis that B-Mode, colour Doppler ultrasonographic measurements and characteristics can identify mastitis in dairy cows. A total of 102 lactating cows were divided into 3 groups: cows in which all mammary quarters were CMT-negative, cows with CMT-positive mammary quarters and cows with clinical mastitis in at least one quarter. Colour Doppler ultrasonography measurements of the supramammary lymph nodes revealed that distortion-type vascular morphology, the rate of type 4 vascular densities and the incidence of mixed-type vascular distributions were highest in the clinical mastitis group, whereas the frequency of avascularity in supramammary lymph nodes was highest in the CMT-negative group. All differences were significant. In conclusion, the use of B-mode and colour Doppler ultrasonographic measurements of the supramammary lymph nodes can provide useful information about the current condition of mastitis in cows, although its diagnostic potential remains to be determined.

In this research communication we used digital gene expression (DGE) analysis to identify differences in gene expression in the mammary glands of dairy cows between early lactation and the mid-dry period. A total of 741 genes were identified as being differentially expressed by DGE analysis. Compared with their expression in dry cows, 214 genes were up-regulated and 527 genes were down-regulated in lactating cow mammary glands. Gene Ontology analysis showed that lactation was supported by increased gene expression related to metabolic processes and nutrient transport and was associated with decreased gene expression related to cell proliferation. Pathway mapping using the Kyoto Encyclopedia of Genes and Genomes showed that 579 differentially expressed genes had pathway annotations related to 204 pathways. Metabolic pathway-related genes were the most significantly enriched. Genes and pathways identified by the present study provide insights into molecular events that occur in the mammary gland between early lactation and mid-dry period, which can be used to facilitate further investigation of the mechanisms underlying lactation and mammary tissue remodeling in dairy cows.

This Research Communication describes the residue concentration of a dry cow antibiotic in two different milk fractions and describes effects of milk fraction and milk composition on the test performance of a rapid screening and a microbial inhibitor test. Thirteen dry cows were treated with an intramammary dry cow antibiotic containing 150 mg cefquinome. Quarter foremilk and stripping samples were collected on the first 10 d postpartum. All milk samples were analyzed for milk composition by the local Dairy Herd Improvement Association and were tested for antibiotic residues using the rapid screening test Milchtest BL and the microbial inhibitor test Delvotest BR Brilliant Plates. The residue concentration of cefquinome was determined in foremilk and stripping samples from milkings 1, 2, 3, 5, and 7 after calving using high performance liquid chromatography – tandem mass spectrometry. The logarithm of cefquinome concentration (logCef) was higher in foremilk than in stripping samples and higher in milk samples with lower lactose content. Furthermore, logCef decreased with the number of milkings (P < 0.001). The Milchtest BL was more likely to be not evaluated (i.e. no test and control line or no control line appeared) in stripping samples and milk samples with higher protein content. In the Delvotest BR Brilliant Plates milk samples with higher protein content were more likely to have a false positive result (i.e. the screening test result was positive, but the HPLC-MS/MS result was below the detection limit of the screening test). These results indicate that foremilk is the recommended milk fraction to be tested for residues of cefquinome and that a high protein content can be a cause of test failure and false positive results when milk during the first 10 d postpartum is tested for antibiotic residues using screening tests.

Dairy cows with ketosis display severe oxidative stress as well as high blood concentrations of non-esterified fatty acids (NEFA) and β-hydroxybutyrate (BHB). Cytochrome P4502E1 (CYP2E1) plays an important role in the induction of oxidative stress. The aim of this study was to investigate CYP2E1 expression and activity in the liver of clinically ketotic cows (in vivo) and the effects of NEFA and BHB on CYP2E1 expression and activity in hepatocytes (in vitro). Dairy cows with clinical ketosis exhibited a low blood concentration of glucose but high concentrations of NEFA and BHB. Hepatic mRNA, protein expression, and activity of CYP2E1 were significantly higher in cows with clinical ketosis than in control cows. In vitro, both NEFA and BHB treatment markedly up-regulated the mRNA and protein expressions as well as activity of CYP2E1 in cow hepatocytes. Taken together, these results indicate that high levels of NEFA and BHB significantly up-regulate the expression and activity of hepatic CYP2E1, and may be influential in the induction of oxidative stress in cows with clinical ketosis.

Automatic cluster remover (ACR) settings regulate the end of milking by detaching the clusters based on milk flow dropping below a preset level, which needs to be standardised for different breeds of dairy animals based on their production. A study was conducted to find out the best ACR setting for milking Indian crossbred cows based on milkability, milking irregularities and milk quality. Fifty six crossbred dairy cows in lactations 1 to 4 were categorised into three groups based on the level of production; low (N = 16; <12 kg/d), medium (N = 32; 12–18 kg/d) and high (N = 08; >18 kg/d). The ACR settings tested were 0.1, 0.2, 0.3 and 0.4 kg/min, keeping the vacuum level and pulsation settings constant. The ACR settings significantly (P < 0.01) affected the milk yield at all levels of production with a significant effect (P < 0.01) on machine-on time at 0.4 kg/min. The yield during the first 2 min of milking, average flow and peak flow rates were not affected at any level of production. The average electrical conductivity in milk was significantly (P < 0.01) lower for the low and medium yield cows without affecting the mean somatic cell count. At 0.4 kg/min, more cluster reattachments were needed because of significant amount of milk remaining in the udders post-cluster removal.

This paper examines ex-ante impacts of two policy interventions that improve productivity of local-breed cows through artificial insemination (AI) and producers’ access to distant markets through a dairy market hub. The majority of cattle in Kilosa district in Tanzania are local low productivity breeds kept by smallholders and agro-pastoralists. Milk production is seasonal, which constrains producers’ access to distant urban markets, constrains producers’ incomes and restricts profitability in dairy processing. We developed and evaluated an integrated system dynamics (SD) simulation model that captures many relevant feedbacks between the biological dynamics of dairy cattle production, the economics of milk market access, and the impacts of rainfall as an environmental factor. Our analysis indicated that in the short (1 year) and medium (5-year) term, policy interventions have a negative effect on producers’ income due to high AI costs. However, in the long term (5+ years), producers’ income from dairy cattle activities markedly increases (by, on average, 7% per year). The results show the potential for upgrading the smallholder dairy value chain in Kilosa, but achievement of this result may require financial support to producers in the initial stages (first 5 years) of the interventions, particularly to offset AI costs, as well as additional consideration of post-farm value chain costs. Furthermore, institutional aspects of dairy market hub have substantial effects on trade-offs amongst performance measures (e.g. higher profit vs. milk consumption at producer's household) with gain in cumulative profit coming at the expense of a proportional and substantial reduction in home milk consumption.

This research paper addresses the hypothesis that comparative genomics can give a new insight into the functionality of casein genes with respect to the casein micelle. Comparative genomics is a rapidly emerging field in computational biology whereby two or more genomes are compared in order to obtain a global view on genomes as well as assigning previously unknown functions for genes. Casein genes are among the most rapidly evolving mammalian genes, with the gene products mainly grouped into four types (αs1-, αs2-, β- and κ-casein). Functionally, casein genes are central to the casein micelle, the exact structure of which is still a subject of intense debate. Moreover, and adding to this complexity, some mammals lack some of the casein genes, although casein micelles have been observed in their milk. This observation has prompted an investigation into the distribution of casein genes across a host of mammalian species. It was apparent from this study that casein gene sequences are very diverse from each other and we confirmed that many mammalian species lack one or more of the casein genes. The genes encoding β- and κ-caseins are present in most mammals whereas α-casein encoding genes are less represented. This suggests different mechanisms for casein micelle formation in different species as well as the functions that are assigned to each individual casein.

The aim of the studies presented in this research communication was to compare species of origin (goat and sheep) and the effect of treatment (pasteurization at 56, 63 and 72 °C, skimming and curding) and refrigeration time on colostrum antimicrobial activity (AnAc). Two experiments were performed. In experiment 1, twenty-four first milking colostrum samples were obtained (12 goats, 12 sheep) and an aliquot of each sample was subjected to 6 different treatments, control (untreated), pasteurization at 56, 63 and 72 °C, skimming and curding. Colostrum AnAc was tested directly against E. coli using disks in a Petri dish and Enrofloxacin (antibiotic) and saline serum as positive and negative control, respectively. Species had no effect (P > 0.05) on colostrum AnAc, and neither did pasteurization at different temperatures or skimming. However, curding showed the lowest colostrum AnAc (P < 0.05) in both species. In the second experiment, four treatments were assayed, control, pasteurization at 56 and 63 °C and skimming. An aliquot of twelve goat colostrum samples were refrigerated after treatments for 10 d at 4 °C. Colostrum AnAc was measured at 0, 2, 4, 6, 8, and 10 d. A reduction in colostrum AnAc was observed due to refrigeration time. The results suggest that if farmers use frozen colostrum for neonates, the process of curding colostrum or refrigeration at 4 °C longer than 4 d is not recommended.

Dairy cows with fatty liver or ketosis display decreased insulin sensitivity and defects in the insulin receptor substrate (IRS)/PI3K/AKT signaling pathway. Phosphatase and tensin homolog (PTEN) is a well-known tumor suppressor and also a negative regulator of insulin signaling and peripheral insulin sensitivity. We investigated the hypothesis that PTEN may affect the insulin pathway-mediated hepatic glucose and lipid metabolism in dairy cows. Adenovirus vectors that over-express and silence PTEN were constructed, and then transfected into hepatocytes isolated from calves to investigate the effect of PTEN on PI3K/AKT signaling pathway. PTEN silencing increased the phosphorylation of AKT and the expression of PI3K but decreased the phosphorylation of IRS1, which increased the phosphorylation levels of glycogen synthase kinase-3β (GSK-3β) and expression of sterol regulatory element-binding protein-1c (SREBP-1c). Increased GSK-3β phosphorylation further up-regulated expression of the key enzymes phosphoenolpyruvate carboxykinase (PEPCK) and glucose-6-phosphatase (G6-Pase) involved in gluconeogenesis. Furthermore, the expression of SREBP-1c target gene fatty acid synthase (FAS) also increased significantly. We further showed that PTEN over-expression could reverse the above results. PTEN negatively regulates the enzymes involved in hepatic gluconeogenesis and lipid synthesis, which suggests that PTEN may be a therapeutic target for ketosis and fatty liver in dairy cows.

The amount of the four caseins (αs1, αs2, β and κ-CN) in donkey milk was evaluated by Urea-PAGE analysis at pH 8.6, followed by immuno-detection with polyclonal antibodies, coupled to densitometric analysis. The results showed the percentage of each casein in decreasing order: β (54.28) > αs1 (35.59) > αs2 (7.19) > κ-CN (2.79). The mRNA quantification of donkey casein transcripts, carried out by RT-qPCR, showed that the average percentage of corresponding gene transcripts (CSN2, CSN1S1, CSN1S2 I and CSN3) was 70.85, 6.28, 14.23 and 8.65, respectively. The observed translation efficiency, assessed as percentage of single milk casein fraction out of single percentage of transcript, was 0.76, 5.66, 0.50 and 0.32, respectively. The analysis of the sequences flanking the start codon, the codon usage frequencies and the coding sequence length might explain, at least in part, the differential transcriptional and translational rate observed among the casein transcripts.

Analysis of milk BHB concentration by Fourier-transform infrared (FTIR) spectrometry more frequently than regular milk testing could help dairy producers in decision making, particularly if it would be possible to use small hand-stripped samples (hereinafter simply called samples) taken between dairy herd improvement (DHI) test-samples analysed using DHI algorithms. The aim of this Research Communication was to evaluate milk BHB concentration and the prevalence of elevated milk BHB concentration analysed by FTIR spectrometry compared with flow-injection analysis (SKALAR) from samples taken at different times relative to the milking. A total of 293 early-lactation cows in 44 commercial dairy herds were involved in the study. Herds were visited once during the morning milking when a routine DHI test-sample was obtained using in-line milk samplers. Additional milk samples were taken by hand stripping as follows: (1) Just before connecting the milking machine; (2) immediately after removing the milking machine; (3) 3 h after milking and (4) 6 h after milking. Milk samples were analysed for BHB concentration by FTIR and SKALAR, the latter being the reference method. Milk BHB concentration from samples taken before milking was different between FTIR and SKALAR whereas no difference was noted for other sampling times, although milk BHB concentration rose as time after milking increased. Except for DHI test-samples for which prevalence was not different between analysis methods, prevalence of elevated milk BHB concentration (≥0.15 mmol/l) was greater for FTIR analysis. However, no difference in prevalence was observed between SKALAR and FTIR when using a threshold of ≥0.20 mmol/l. In summary, hand-stripped milk samples taken any time after removing the milking machine until 6 h after the milking can be recommended for FTIR analysis of elevated milk BHB concentration prevalence provided a threshold of 0.20 mmol/l is used.

We investigated whether variation of the sheep Growth Hormone Receptor (GHR), Growth Hormone Releasing Hormone Receptor (GHRHR) and Insulin-Like Growth Factor 1 (IGF1) genes were associated with milk coagulation properties (MCP) in sheep. The GHR, GHRHR and IGF1 genes are part of the GH system, which is known to modulate metabolism, growth and reproduction as well as mammogenesis and galactopoiesis in dairy species. A total of 380 dairy Sarda sheep were genotyped for 36 SNPs mapping to these three genes. Traditional MCP were measured as rennet coagulation time (RCT), curd-firming time (k20) and curd firmness at 30 m (a30). Modeling of curd firming over time (CFt) was based on a 60 m lactodynamographic test, generating a total of 240 records of curd firmness (mm) for each milk sample. The model parameters obtained included: the rennet coagulation time as a result of modeling all data available (RCTeq, min); the asymptotic potential value of curd firmness (CFP, mm) at an infinite time; the CF instant rate constant (kCF, %/min); the syneresis instant rate constant (kSR, %/min); the maximum value of CF (CFmax, mm) and the time at achievement of CFmax (tmax, min). Statistical analysis revealed that variation of the GHR gene was significantly associated with RCT, kSR and CFP (P < 0.05). No other significant associations were detected. These findings may be useful for the dairy industry, as well as for selection programs.

We hypothesised that a relationship would exist between hair fatty acids, especially C12:0, C14:0 and C16:0, and parameters of energy metabolism such as energy intake, energy mobilisation, and energy requirement for maintenance and milk performance. For this study, 11 primiparous German Holstein cows were available from which hair samples at weeks 6 and 8 of lactation were analysed. The average body weight of these animals was 558 ± 27 kg at calving and milk yield at 100-days in milk was 3,537 ± 529 kg. Feed intake and milk yield were measured daily. Body weight and back fat thickness were measured at calving and in weeks 2, 4, and 8 of lactation. Energy balance and energy utilisation were calculated until week 6 of lactation. Spearman's correlation coefficients were found to be significantly positive for the relationship between the percentage of C12:0 and C14:0 fatty acids in the hair in lactation week 8 and energy intake in weeks 5 and 6 (0.62 < r < 0.65, P < 0.05). If the animals are grouped according to their energy utilisation between weeks 1 and 6 into two groups higher (n = 6) or lower (n = 5) than the median, animals of the high energy utilising group had a higher energy intake. These animals had also higher percentages of the C12:0 fatty acid in their hair fat (week 6: 4.9% vs. 3.1%, P < 0.05; week 8: 4.3% vs. 2.9%, P = 0.05). Our hypothesis is supported, and this study justifies further investigation of the content of medium-chain fatty acids in hair samples as biomarkers for the metabolic status of a cow during early lactation.

This research was carried out to quantify the effects of a range of variables on milk fat globule (MFG) size for a herd of Holstein-Friesian cows managed through an automatic milking system with year-round calving. We hypothesised that the overall variation in average MFG size observed between individual animals of the same herd cannot sufficiently be explained by the magnitude of the effects of variables that could be manipulated on-farm. Hence, we aimed to conduct an extensive analysis of possible determinants of MFG size, including physiological characteristics (parity, days in milk, days pregnant, weight, age, rumination minutes, somatic cell count) and milk production traits (number of milkings, milk yield, fat yield, protein and fat content, fat-protein ratio) on the individual animal level; and environmental conditions (diet, weather, season) for the whole herd. Our results show that when analysed in isolation, many of the studied variables have a detectable effect on MFG size. However, analysis of their additive effects identified days in milk, parity and milk yield as the most important variables. In accordance with our hypothesis, the estimated effects of these variables, calculated using a multiple variable linear mixed model, do not sufficiently explain the overall variation between cows, ranging from 2.70 to 5.69 µm in average MFG size. We further show that environmental variables, such as sampling day (across seasons) or the proportion of pasture and silage in the diet, have limited effects on MFG size and that physiological differences outweigh the effects of milk production traits and environmental conditions. This presents further evidence that the selection of individual animals is more important than the adjustment of on-farm variables to control MFG size.

The aim of this work was to use X-ray diffraction to identify substances used for adulteration of raw milk and to determine if crystallographic analysis can detect extraneous substances in milk. Two unknown substances were sent anonymously by employers linked to the dairy chain, who claimed that they were added directly in milk prior to water addition by truck drivers. The samples were analyzed by X-ray diffraction and submitted to physicochemical analysis. The first substance was identified by X-ray diffraction as sodium citrate, complying with its physicochemical attributes, such as the powerful ability to decrease the freezing point. The second substance was identified by X-ray diffraction as sucrose and this result was also in agreement with its ability to increase the density, decrease the freezing point and finally, to be positive for sucrose in the resorcinol qualitative test. To evaluate if X-ray diffraction can detect extraneous substances already mixed in milk, fresh raw milk samples tampered with urea, sodium hydroxide, sodium citrate and sucrose were freeze dried and analyzed by X-ray diffraction, with no detection of any extraneous substances at any percentage. This is the first report of attempted diagnosis of extraneous substances in milk by X-ray diffraction. However, this technique can be useful only when applied to identify substances used for adulteration prior to its dilution in milk, since the amorphous nature of milk seems to be a limitation for the accurate detection of extraneous substances.

This research communications addresses the hypothesis that a part of iso 17:0 and anteiso 17:0 in milk fat could come from endogenous extraruminal tissue synthesis. In order to confirm this a linear regression model was applied to calculate the proportions of iso 17:0 and anteiso 17:0 in milk fat that could come from elongation of their putative precursors iso 15:0 and anteiso 15:0, respectively. Sixteen dairy goats were allocated to two simultaneous experiments, in a crossover design with four animals per treatment and two experimental periods of 25 d. In both experiments, alfalfa hay was the sole forage and the forage to concentrate ratio (33 : 67) remained constant. Experimental diets differed on the concentrate composition, either rich in starch or neutral detergent fibre, and they were administered alone or in combination with 30 g/d of linseed oil. Iso 15:0, anteiso 15:0, iso 17:0 and anteiso 17:0, the most abundant branched-chain fatty acids in milk fat, were determined by gas chromatography using two different capillary columns. The regression model resolved that 49% of iso 17:0 and 60% of anteiso 17:0 in milk fat was formed extraruminally from iso 15:0 and anteiso 15:0 elongation.

This research communication describes how different detachment levels (0.48, 0.3 and 0.06 kg milk/min) at the quarter-level affect milk flow profiles and overall milking efficiency in automatic milking systems. We hypothesized a higher detachment level would result in greater mean flow rates without affecting the volume of harvested milk per cow during 24 h compared to lower detachment levels. The data suggest milk flow decreased to a rate below the overmilking limit within the 6-s delay time required for termination in all treatments, but the duration of overmilking was shorter for the greatest detachment level compared to the other treatments. We conclude that setting a detachment level at a greater milk flow rate reduces the duration of overmilking without affecting the amount of milk harvested when applied to cows in mid-lactation during quarter-level milking. We also suggest that the steepness of the decline phase of the milk flow curve might have a larger effect than the actual detachment level on the duration of overmilking.

The objectives of the research described here were to describe the persistence of intramammary infections (IMI) caused by coagulase negative staphylococci (CNS) in goats using strain-typing, and to evaluate the relationship between species-specific CNS IMI and somatic cell score (SCS) at the udder-half level. Udder-half milk samples were collected from all 909 lactating goats (1817 halves; 1 blind half) in a single herd. Milk samples were cultured on Columbia blood agar, and 220 goats with at least one half yielding a single colony type CNS were enrolled for two additional half-level samplings at approximately 1-month intervals. Isolates were identified to the species level by matrix-assisted laser desorption-ionisation time-of-flight mass spectrometry or PCR amplification and partial sequencing of tuf or rpoB. An IMI was defined as persistent when ≥1 follow-up sample yielded the same species and strain as on Day 0 based on pulsed-field gel electrophoresis. A generalised mixed linear model was used to evaluate the odds of persistence as a function of CNS species. A mixed linear model was used to evaluate the relationship between IMI status on a given day and SCS. Among 192 IMI, 69.8% were persistent based on species and strain-type. Staphylococcus simulans IMI had higher odds of persistence than Staphylococcus arlettae IMI. In primiparous goats, Staphylococcus epidermidis IMI was associated with higher SCS than S. arlettae, Staphylococcus xylosus and ‘other CNS’ IMI. The differences detected in the present study between CNS species, with regard to persistence of IMI and association with SCS, highlight the need to study CNS at the species and strain level to understand the pathogenicity and epidemiology of CNS in goats.