Introduction: The aim of this study was to investigate the relationship between body condition score (BCS) determined on the dry-off day, calving day, and in the first month of lactation, its changes during the dry period and early lactation, and periparturient indices and fertility in high-producing dairy cows. Material and Methods: The experiment was conducted in two herds: A and B, located in Western Poland. The studies were conducted on 116 and 108 Polish Holstein-Friesian dairy cows respectively, with an average milk yield of >10 000 kg/305-day lactation. The experiment included the dry period (-56 d to the calving day), the calving day, and early lactation (from +1 to +56 d). The experimental factor was BCS (0 to 5-point scale). The BCS was performed by one person on day -56, on parturition day (in the first 12 h after calving) and on day 30 of lactation. Results: A decrease in BCS (≥-0.25) in herd A during the dry period accelerated the planned calving period by 7.3 d. In the group of cows with BCS <3.25 on the dry-off day, the lowest artificial insemination index (1.80), the shortest period of insemination services (25 d), and days open (87 d) were recorded. Moreover, cows with BCS < 3.25 at calving had the shortest days open (91 d). BCS >3.50 in the first month of lactation (30 d) resulted in the extension of uterine involution period (56 d). Improvement of BCS during the dry period shortened the anoestrus (60 d) in herd A and the period of insemination service (60 d) in herd B. However, in this group (IM BCS ≥ 0.25) of cows the day of the highest artificial insemination index (2.50) in herd B was analysed. Conclusion: The body condition on the dry-off day and at calving, as well as its deterioration in the first month of lactation, have a considerable effect on fertility indices in dairy cows, thus confirming the advisability of its regular monitoring during routine operations connected with the management of a dairy cattle herd.

Introduction: The aim of this study was to investigate the relationship between body condition score (BCS) determined on the dry-off day, calving day, and in the first month of lactation, its changes during the dry period and early lactation, and periparturient indices and fertility in high-producing dairy cows. Material and Methods: The experiment was conducted in two herds: A and B, located in Western Poland. The studies were conducted on 116 and 108 Polish Holstein-Friesian dairy cows respectively, with an average milk yield of >10 000 kg/305-day lactation. The experiment included the dry period (-56 d to the calving day), the calving day, and early lactation (from +1 to +56 d). The experimental factor was BCS (0 to 5-point scale). The BCS was performed by one person on day -56, on parturition day (in the first 12 h after calving) and on day 30 of lactation. Results: A decrease in BCS (≥-0.25) in herd A during the dry period accelerated the planned calving period by 7.3 d. In the group of cows with BCS <3.25 on the dry-off day, the lowest artificial insemination index (1.80), the shortest period of insemination services (25 d), and days open (87 d) were recorded. Moreover, cows with BCS < 3.25 at calving had the shortest days open (91 d). BCS >3.50 in the first month of lactation (30 d) resulted in the extension of uterine involution period (56 d). Improvement of BCS during the dry period shortened the anoestrus (60 d) in herd A and the period of insemination service (60 d) in herd B. However, in this group (IM BCS ≥ 0.25) of cows the day of the highest artificial insemination index (2.50) in herd B was analysed. Conclusion: The body condition on the dry-off day and at calving, as well as its deterioration in the first month of lactation, have a considerable effect on fertility indices in dairy cows, thus confirming the advisability of its regular monitoring during routine operations connected with the management of a dairy cattle herd.

Introduction: Bovine postpartum metritis causes great losses. Mast cell (MC)-released mediators participate in uterine inflammation and immune response, but their role in postpartum metritis in cows has not been reported. This study investigated the effect of endometrial MC on the disorder.Material and Methods: Ten dairy cows, at 6 to 10 days postpartum and with acute purulent metritis made up the experimental group, and 10 comparable healthy cows the control group. Endometrial histamine and IgE levels were determined by ELISA, and the MC particle state and expression of histamine H₁ (H₁R) and H₂ (H₂R) mRNA receptors were examined by transmission electron microscope and real-time quantitative PCR, respectively.Results: Endometrial histamine and IgE levels were significantly higher in the experimental group. In the control group, homogenously distributed size-varied granules were seen in MC cytoplasm of endometrium of lamina propria. In the experimental group however, these showed degranulation with features of reduction. The level of H₁R mRNA was lower in the experimental group, but that of H₂R mRNA was higher.Conclusion: The results suggest MC type I hypersensitivity characteristics during metritis, and histamine provocation of local inflammation. High expression of H₂R and low expression of H₁R inhibited the inflammatory response and prevented excessive uterine tissue damage.

Introduction: Bovine postpartum metritis causes great losses. Mast cell (MC)-released mediators participate in uterine inflammation and immune response, but their role in postpartum metritis in cows has not been reported. This study investigated the effect of endometrial MC on the disorder.

The aim of this study was to determine changes of reactive oxygen species (ROS), serum antioxidant capacity (SAC), oxidative stress index (OSi), and α-tocopherol (α-T) during the periparturient period in healthy and mastitic cows and to further investigate whether these parameters can be used as a tool for identifying cows at higher risk of developing mastitis. Blood samples from 110 dairy cows from two commercial farms were obtained at dry-off, calving, and 30 days post-partum. Healthy cows formed group A (n = 90) and mastitic cows B (n = 20). Blood serum was obtained by centrifugation, and the aforementioned parameters were determined. A general linear model was used for analysing the associations among the determined blood parameters, the health of the animals’ udder, and the sampling time. ROS and OSi values were higher (P < 0.001) by a respective 14% and 26%, and SAC values lower (P < 0.001) by 10% in group B than in group A at calving. ROC curve analysis revealed that all determined parameters at calving and α-T at dry-off and 30 days post-partum had excellent or acceptable predicting ability for mastitis incidence. This information provides a tool for early identification of cows at high risk of developing mastitis, allowing the implementation of intervention strategies.

The aim of this study was to determine changes of reactive oxygen species (ROS), serum antioxidant capacity (SAC), oxidative stress index (OSi), and α-tocopherol (α-T) during the periparturient period in healthy and mastitic cows and to further investigate whether these parameters can be used as a tool for identifying cows at higher risk of developing mastitis.

The objectives of this study were to determine the role of a fall in pre-calving body condition score (BCS) in postpartum metabolic status and reproductive outcomes, and gauge the indicativeness of blood metabolites during the transition period. Cows were grouped based on BCS loss between days −14 ±3 and 0 relative to calving. Cows that lost no BCS were the BCS control group (BCS-C), cows that lost 0.25 BCS points the low BCS loss group (BCS-L), and those that lost 0.5 points or more the high BCS loss (BCS-H) group. Blood was taken on days −14 ±3, 3, 14, and 30 ±4 for determination of comprehensive metabolic panel biomarker levels. Beta-hydroxybutyric acid (BHBA) levels were quantified on postpartum examination days. Vaginal discharge scores, ovarian activity on day 30 ±4, and subsequent fertility parameters were evaluated. The BCS-H cows had lower mean Ca concentrations before calving and on day 3, when the group’s BHBA and CK were higher (P < 0.05); on day 14 they had higher AST concentrations (P < 0.05). The BCS-L cows had greater bilirubin levels (P < 0.05). The BCS-H cows had lower cyclicity and higher endometritis rates. First service pregnancy rates were 50%, 50%, and 61.9%, open days 96.8, 95.75, and 89.2, and overall pregnancy rates 56.25%, 65%, and 80.95 % in the BCS-H, BCS-L, and BCS-C groups, respectively. Prepartum BCS loss of ≥ 0.5 points could be associated with Brown Swiss cow low Ca and BHBA concentrations early postpartum, and with subsequent uterine health and overall pregnancy rate. Prepartum Ca concentration might be a prognostic biomarker for postpartum metabolic status and reproductive outcomes.

The objectives of this study were to determine the role of a fall in pre-calving body condition score (BCS) in postpartum metabolic status and reproductive outcomes, and gauge the indicativeness of blood metabolites during the transition period.

This study aimed to characterise the effects of ketosis on milk yield and composition and digestive capacity in transition dairy cows. Seven ketotic and seven healthy cows were housed in individual stalls for six days. Samples of plasma, milk, refused total mixed ration, and faeces were collected, and the blood biochemical parameters, milk yield and composition, dry matter intake, and faecal dry matter (FDM) production were determined. Compared with healthy cows, the ketotic cows had significantly higher concentrations of milk fat and citrate, but lower levels of milk protein and lactose. The cows exhibited a need for acid detergent fibre in forage and better digestion of neutral detergent fibre, starch, crude protein, and phosphorus than healthy cows, but more fat and gross energy were excreted in their faeces. Ketotic cows had higher energy-corrected milk yields and lower FDM than healthy cows. Lower feed intake coinciding with the requirement to maintain high milk production is considered to be the cause of ketosis in dairy cows. Ketotic cows exhibited lower dry matter fat digestion.

This study aimed to characterise the effects of ketosis on milk yield and composition and digestive capacity in transition dairy cows.

Introduction: There are many veterinary products containing β-lactam antibiotics which are used for mastitis treatment in cows. The aim of the study was to determine whether mastitis could have any effect on amoxicillin (AMX) or penicillin G procaine (PEN) withdrawal period from milk, in the context of current maximum residue limits established by the European Commission. Material and Methods: The study was conducted on 17 dairy Black and White cows with clinical mastitis during the lactation period. The first group (n = 8) received 200 mg of amoxicillin (AMX), whereas the second group (n = 9) received 200,000 IU/mg of penicillin G procaine (PEN) by intramammary administration. For the measurement of AMX and PEN concentrations in milk, the liquid chromatography tandem mass spectrometry method was applied. Pharmacokinetic calculations were performed using Phoenix WinNonlin 6.4 software. Results: The determined AMX and PEN half-life values in the mammary gland suggest that the drug withdrawal is at a level of 99.9% within 81 h (≈3.5 days) and 116 h (≈5 days) after administration of AMX and PEN, respectively. The present research indicates that, at 60 h after administration, the average PEN concentration in the milk from cows with clinical signs of mastitis may still reach 4.96 g/kg and that of AMX can even be 6.92 g/kg. Conclusion: The results obtained confirm that, in mastitis cases, a 72-h withdrawal period is sufficient for elimination of AMX to a lower level than the established maximum residue limit (MRL) values. However, in the case of PEN, at 69 h after administration, the drug concentration may be close to that of the determined MRL.

Introduction: There are many veterinary products containing β-lactam antibiotics which are used for mastitis treatment in cows. The aim of the study was to determine whether mastitis could have any effect on amoxicillin (AMX) or penicillin G procaine (PEN) withdrawal period from milk, in the context of current maximum residue limits established by the European Commission.

Introduction: Vascular cell adhesion molecule 1 (VCAM-1) is a member of Ig superfamily. The aim of this study was to prepare highly specific polyclonal antibodies against bovine VCAM-1 and to evaluate the expression of VCAM-1 in the mammary lymph nodes of cows with subclinical mastitis.Material and Methods: The VCAM-1 gene was cloned from bovine Peyer’s patches and inserted into the pGEX-4T-1 and pET-28a vectors. The recombinant plasmids pGEX-4T-1/VCAM-1 and pET-28a/VCAM-1 were transferred into Escherichia coli BL21 and the recombinant strains were induced by isopropyl-D-thiogalactoside to produce fusion proteins tagged with polyhistidine (His) and glutathione S-transferase (GST), respectively. The expressed fusion proteins His-VCAM-1 and GST-VCAM-1 were identified by SDS-PAGE and Western blot. His-VCAM-1 protein was used as an antigen to immunise Wistar rats and polyclonal antibody serum against VCAM-1 was obtained.Results: The serum titre tested by indirect ELISA was 128,000 using GST-VCAM-1 as the well coating antigen. Western blots indicated that the antibody recognised recombinant VCAM-1 protein as well as endogenous VCAM-1. In addition, using qPCR and Western blot, VCAM-1 mRNA and protein expression levels were measured in dairy cows with subclinical mastitis. It was demonstrated that VCAM-1 levels in the mammary lymph nodes of the cows were significantly higher than those from healthy controls (P < 0.05).Conclusion: These results are to our knowledge the first report that VCAM-1 expression in the mammary lymph nodes is elevated in dairy cows with subclinical mastitis.

Introduction: Vascular cell adhesion molecule 1 (VCAM-1) is a member of Ig superfamily. The aim of this study was to prepare highly specific polyclonal antibodies against bovine VCAM-1 and to evaluate the expression of VCAM-1 in the mammary lymph nodes of cows with subclinical mastitis.

Introduction: The aim of the study was to investigate changes in the serum levels of adiponectin and TNF-α, as well as insulin sensitivity, and to elucidate the possible relationship among the parameters and negative energy balance during the periparturient period of dairy cows.Material and Methods: Thirty primiparous Holstein dairy cows were selected for the study. Blood samples were collected from each cow seven days before the expected calving date, on the calving day, and 7, 14, and 21 days after calving. Blood non-esterified fatty acids (NEFA), β-hydroxybutyric acid (BHBA), glucose, insulin, adiponectin, and TNF- α levels were measured. Revised Quantitative Insulin Sensitivity Check Index (rQUICKI) was calculated using data on NEFA, insulin, and glucose concentrations.Results: When compared to prepartum levels, serum concentration of adiponectin significantly increased on day 21 postpartum. The rQUICKI increased and NEFA levels decreased on day 7 after parturition. Insulin and glucose levels decreased on days 7, 14, and 21 postpartum when compared with prepartum levels. BHBA levels decreased on day 21 and TNF- α concentration also decreased on days 7, 14, and 21 postpartum. Adiponectin levels positively correlated with NEFA during the preparturient period. Negative correlation was detected between adiponectin and rQUICKI on calving day and on 14ᵗʰ day after parturition. TNF- α concentration positively correlated with glucose levels on day 7 prepartum and on 21ˢᵗ day postpartum and with rQUICKI on 21ˢᵗ day postpartum. Negative correlation was detected between adiponectin level and insulin sensitivity.Conclusion: Based on the results of the study, we concluded that adiponectin could possibly increase insulin sensitivity when blood NEFA concentrations are elevated.

Introduction: The aim of the study was to investigate changes in the serum levels of adiponectin and TNF-α, as well as insulin sensitivity, and to elucidate the possible relationship among the parameters and negative energy balance during the periparturient period of dairy cows.

Introduction: To identify novel pathways involved in the pathogenesis of ketosis, an isobaric tag for relative and absolute quantitation/mass spectrometry was used to define differences in protein expression profiles between healthy dairy cows and those with clinical or subclinical ketosis.Material and Methods: To define the novel pathways of ketosis in cattle, the differences in protein expression were analysed by bioinformatics. Go Ontology and Pathway analysis were carried out for enrich the role and pathway of the different expression proteins between healthy dairy cows and those with clinical or subclinical ketosis.Results: Differences were identified in 19 proteins, 16 of which were relatively up-regulated while the remaining 3 were relatively down-regulated. Sorbitol dehydrogenase (SORD) and glyceraldehyde-3-phosphate dehydrogenase (G3PD) were up-regulated in cattle with ketosis. SORD and G3PD promoted glycolysis. These mechanisms lead to pyruvic acid production increase and ketone body accumulation.Conclusion: The novel pathways of glycolysis provided new evidence for the research of ketosis.

Introduction: To identify novel pathways involved in the pathogenesis of ketosis, an isobaric tag for relative and absolute quantitation/mass spectrometry was used to define differences in protein expression profiles between healthy dairy cows and those with clinical or subclinical ketosis.