Blood Of Cows Research Articles

Metabolic Fingerprinting of Blood and Urine of Dairy Cows Affected by Bovine Leukemia Virus: A Mass Spectrometry Approach.

This study investigated metabolic changes associated with bovine leukemia virus (BLV) infection in dairy cows, focusing on pre-parturition alterations. Metabolite identification in serum and urine samples was performed using a targeted metabolomics method, employing the TMIC Prime kit in combination with flow injection analysis and liquid chromatography-tandem mass spectrometry. Of 145 cows examined, 42 (28.9%) were BLV-seropositive. Around 38% of infected cows showed high somatic cell counts indicative of subclinical mastitis, with 15 experiencing additional health issues such as ketosis, milk fever, and lameness. Despite these conditions, no significant differences in milk yield or composition were observed between the infected and control groups. Metabolomic analysis conducted at -8 and -4 weeks prepartum revealed significant metabolic differences between BLV-infected and healthy cows. At -8 weeks, 30 serum metabolites were altered, including sphingomyelins, lysophosphatidylcholines, amino acids, and acylcarnitines, suggesting disruptions in membrane integrity, energy metabolism, and immune function indicative of early neoplastic transformations. By -4 weeks, the number of altered metabolites decreased to 17, continuing to reflect metabolic disruptions in cows with leukemia. Multivariate analysis highlighted distinct metabolic profiles between infected and control cows, identifying key discriminating metabolites such as choline, aspartic acid, phenylalanine, and arginine. Urine metabolomics revealed significant prepartum shifts in metabolites related to glucose, asymmetric dimethylarginine, and pyruvic acid, among others. The research confirmed metabolomics' efficacy in defining a BLV infection metabolic profile, elucidating leukosis-associated metabolic disruptions. This approach facilitates the identification of BLV-infected cows and enhances understanding of infection pathophysiology, providing a foundation for advanced management and intervention strategies in dairy herds. The study underscores the profound impact of leukosis on metabolic processes and highlights urine metabolomics' utility in non-invasively detecting BLV infection, offering the potential for improved herd health management.

Transcriptome analysis identifies the NR4A subfamily involved in the alleviating effect of folic acid on mastitis induced by high concentration of Staphylococcus aureus lipoteichoic acid

BackgroundStaphylococcus aureus (S. aureus) mastitis results in economic losses during dairy production. Understanding the biological progression of bovine S. aureus mastitis is vital for its prevention. Lipoteichoic acid is a key virulence factor of S. aureus (aLTA), but the main biological pathways involved in its effect on bovine mammary epithetionallial cells (Mac-T) apoptosis and necrosis have not been fully explored. Folic acid (FA) has anti-inflammatory and anti-apoptotic effects. However, the role of FA in mediating the effects of aLTA on apoptosis and necrosis remains unknown.ResultsWe found that low concentration of aLTA inhibited apoptosis and necrosis and that high concentration promoted the apoptosis and necrosis of Mac-T. FA pretreatment alleviated high concentration of aLTA induced apoptosis. Through transcriptomic analysis, we found that nuclear receptor subfamily 4 group A (NR4A), which alters the expression of downstream genes involved in apoptosis, proliferation, and inflammation, decreased under stimulation with a low concentration of aLTA and increased under stimulation with a high concentration of aLTA. Under stimulation with a high concentration of aLTA, the expression of the NR4A subfamily could be inhibited by FA. The results showed that aLTA may affect apoptosis and necrosis through the NR4A subfamily by targeting genes involved in bacterial invasion of epithelial cells, the IL-17 signaling pathway, DNA replication, longevity regulation, the cell cycle, and tight junction pathways. We further found that the expression trends of NR4A1 and the target genes of the NR4A subfamily (PTGS2, ESPL1, MCM5, and BUB1B) in the blood of healthy cows (Healthy), subclinical mastitis cows (SCM), and SCM supplemented with FA (SCM_FA) were consistent with those observed at the cellular level in this study.ConclusionsOur study revealed that low and high concentrations of aLTA have opposite effects on apoptosis and necrosis of Mac-T and that FA can alleviate the apoptosis induced by high concentration of aLTA. Transcriptome analysis revealed that the NR4A subfamily play a role in the ability of FA to alleviate the apoptosis and necrosis induced by high concentration of aLTA.

Acute phase responses in clinically healthy multiparous Holsteins with and without calcium dysregulation during the early postpartum period

Patterns of calcium dysregulation resulting in low total serum calcium concentrations (tCa) at 4 DIM, known as dyscalcemia, commonly occur in multiparous Holsteins. Dyscalcemia is associated with risk of disease, decreased production, and poor reproductive performance. Inflammation is well-documented early in lactation and is associated with similarly suboptimal outcomes. The acute phase response produces markers and mediators of inflammation; therefore, the objective of our case-control study was to evaluate postpartum patterns of 3 positive acute phase proteins in cows with and without dyscalcemia. We hypothesized that dyscalcemic cows would experience more activated inflammation than eucalcemic cows and that inflammation would precede dyscalcemia diagnosis. Multiparous Holstein cows at 2 commercial dairy farms in central New York were enrolled from a parent study based on tCa at 4 DIM, at a 2:1 ratio of eucalcemic (tCa >2.3 mmol/L; n = 32) to dyscalcemic cows (tCa <2.2 mmol/L; n = 16). Blood was collected 1 to 3 d before parturition and once every 24 h postpartum through 4 DIM. Samples were analyzed for 3 acute phase proteins, serum amyloid A (SAA), haptoglobin and lipopolysaccharide binding protein (LBP). Patterns of protein concentrations in blood over time were compared using linear mixed effects models including fixed effects of calcium status group, time, parity group, farm, relevant 2-way interactions, and the random effect of cow. Overall, dynamics of acute phase proteins showed that dyscalcemic cows experienced increased acute phase responses compared with eucalcemic cows, and that these responses preceded dyscalcemia diagnosis at 4 DIM. Dyscalcemic cows had elevated concentrations of SAA beginning at 2 DIM (eucalcemic: mean = 13.88 µg/mL, 95% CI = 11.34 to 16.99 µg/mL; dyscalcemic: mean = 32.95 µg/mL, 95% CI = 24.55 to 44.21 µg/mL). and continuing through 4 DIM (eucalcemic: mean = 8.14 µg/mL, 95% CI = 6.66 to 9.95 µg/mL; dyscalcemic: mean = 30.01 µg/mL, 95% CI = 22.60 to 39.83 µg/mL). Haptoglobin concentrations were also elevated in the blood of dyscalcemic cows from 2 DIM (eucalcemic: mean = 0.39 g/L, 95% CI = 0.31 to 0.49 g/L; dyscalcemic: mean = 1.11 g/L, 95% CI = 0.79 to 1.56 g/L) through 4 DIM (eucalcemic: mean = 0.27 g/L, 95% CI = 0.21 to 0.34 g/L; dyscalcemic: mean = 1.65 g/L, 95% CI = 1.19 to 2.28 g/L). Concentrations of LBP exhibited a different pattern with a small difference between groups at 3 DIM (eucalcemic: mean = 4.67 µg/mL, 95% CI = 4.02 to 5.42 µg/mL; dyscalcemic: mean = 7.91 µg/mL, 95% CI = 6.49 to 9.63 µg/mL) that became larger at 4 DIM (eucalcemic: mean = 4.88 µg/mL, 95% CI = 4.22 to 5.64 µg/mL; dyscalcemic: mean = 10.79 µg/mL, 95% CI = 8.84 to 13.17 µg/mL). Our work supports the hypothesis that dyscalcemia and inflammatory activity are associated in dairy cows under naturally occurring postpartum conditions. While the causal structure of this relationship remains unknown, improved understanding of inflammation and dyscalcemia may provide insight into mechanisms by which some cows experience maladaptation during the early postpartum period.

Digital Holographic Microscopy in Veterinary Medicine-A Feasibility Study to Analyze Label-Free Leukocytes in Blood and Milk of Dairy Cows.

For several years, the determination of a differential cell count of a raw milk sample has been proposed as a more accurate tool for monitoring the udder health of dairy cows compared with using the absolute somatic cell count. However, the required sample preparation and staining process can be labor- and cost-intensive. Therefore, the aim of our study was to demonstrate the feasibility of analyzing unlabeled blood and milk leukocytes from dairy cows by means of digital holographic microscopy (DHM). For this, we trained three different machine learning methods, i.e., k-Nearest Neighbor, Random Forests, and Support Vector Machine, on sorted leukocyte populations (granulocytes, lymphocytes, and monocytes/macrophages) isolated from blood and milk samples of three dairy cows by using fluorescence-activated cell sorting. Afterward, those classifiers were applied to differentiate unlabeled blood and milk samples analyzed by DHM. A total of 70 blood and 70 milk samples were used. Those samples were collected from five clinically healthy cows at 14-time points within a study period of 26 days. The outcome was compared with the results of the same samples analyzed by flow cytometry and (in the case of blood samples) also to routine analysis in an external laboratory. Moreover, a standard vaccination was used as an immune stimulus during the study to check for changes in cell morphology or cell counts. When applied to isolated leukocytes, Random Forests performed best, with a specificity of 0.93 for blood and 0.84 for milk cells and a sensitivity of 0.90 and 0.81, respectively. Although the results of the three analytical methods differed, it could be demonstrated that a DHM analysis is applicable for blood and milk leukocyte samples with high reliability. Compared with the flow cytometric results, Random Forests showed an MAE of 0.11 (SD = 0.04), an RMSE of 0.13 (SD = 0.14), and an MRE of 1.00 (SD = 1.11) for all blood leukocyte counts and an MAE of 0.20 (SD = 0.11), an RMSE of 0.21 (SD = 0.11) and an MRE of 1.95 (SD = 2.17) for all milk cell populations. Further studies with larger sample sizes and varying immune cell compositions are required to establish method-specific reference ranges.

Vampire bats rapidly fuel running with essential or non-essential amino acids from a blood meal.

In most mammals, running is fuelled by oxidization of endogenous carbohydrates and lipids while amino acids contribute little (< 5-10%). Common vampire bats (Desmodus rotundus), however, specialize on a unique, protein-rich blood diet. Therefore, we hypothesized that (i) vampire bats would rapidly begin utilizing dietary amino acids to support running metabolism, and (ii) that relative reliance on essential and non-essential amino acids would be similar. We fed bats cow's blood enriched either with isotopically labelled glycine (non-essential amino acid) or leucine (essential amino acid). Bats were exercised at speeds of 10, 20 and 30 m min-1 on a respirometry treadmill, allowing us to assess metabolic rate (i.e. O2 consumption and CO2 production) and track the oxidation of labelled amino acids in exhaled CO2. Vampire bats oxidized amino acids as their primary fuel as indicated by a respiratory exchange ratio (RER = ratio of CO2 production to O2 consumption rates) of approximately 0.8-0.9 at all speeds, with the labelled meal accounting for as much as 60% of oxidized fuels at peak usage. Similar oxidation rates indicated bats did not discriminate between essential and non-essential amino acid use. These findings reiterate how strongly metabolism can be shaped by a specialized diet.

Effect of parenteral trace element supplementation on oxidative stress and transcriptomic profile of peripheral blood in peripartum dairy cows

Effect of parenteral trace element supplementation on oxidative stress and transcriptomic profile of peripheral blood in peripartum dairy cows

Short-term reduction in feed intake by dairy cows in the postpartum period leads to subclinical ketosis development

Subclinical ketosis is widespread in highly productive dairy cows after calving and often remains undiagnosed, leading to reduced productivity. Physiologically controlled feeding in the first weeks after calving and during the intensive lactation period can reduce the incidence of ketosis. The study aimed to determine how a short-term reduction of feed after calving affects the formation of ketone bodies in blood, urine and milk of dairy cows. The group of ten Ukrainian black-spotted dairy breed cows after calving aged from 4 to 6 years was involved in the experiment lasted for 72 h. In 24 h the amount of compound feed, haylage, and silage received by cows was gradually reduced until complete exclusion in the diet. Blood, urine, and milk samples were collected three times a day. The content of ketone bodies, β-hydroxybutyrate and glucose was estimated with the corresponding sets of indicator strips. The content of total bilirubin, cholesterol, albumin and enzymes activity in the blood serum were determined on biochemical analyzer. It was found that in 24 h after the beginning of feed reduction the level of blood glucose decreased, persistent hypoglycemia within 48 and 72 h was developed, the concentration of β-hydroxybutyrate in blood and milk and of ketone bodies in urine was elevated. The increase in total bilirubin concentration and liver enzymes activity in the blood serum with a simultaneous decrease in albumin level and delayed sodium propionate conversion into glucose were observed. Thus, short-term reduction in feed intake by dairy cows after calving causes ketosis development and violation of liver functions. Keywords: dairy сows, feeding level, glucose, ketosis, liver damage, β-hydroxybutyrate

The influence of probiotic administration on selected leukocyte subpopulations and the serum amyloid A concentration in the peripheral blood of dairy cows during different lactation periods

The influence of probiotic administration on selected leukocyte subpopulations and the serum amyloid A concentration in the peripheral blood of dairy cows during different lactation periods

Investigating the IgM and IgG B Cell Receptor Repertoires and Expression of Ultralong Complementarity Determining Region 3 in Colostrum and Blood from Holstein-Friesian Cows at Calving.

In cattle, colostral maternal immunoglobulins and lymphocytes transfer across the neonate's intestinal epithelium to provide protection against pathogens. This study aimed to compare repertoires of B cell populations in blood and colostrum in cows for the first time, with an emphasis on ultralong complementarity determining region 3 (CDR3, ≥40 amino acids). Blood mononuclear cells (BMCs, n= 7) and colostral cells (n = 7) were isolated from Holstein-Friesian dairy cows. Magnetic-activated cell sorting was used to capture IgM and IgG B cells from BMCs. Colostral cells were harvested by centrifugation. RNA was extracted and cDNA was produced; IgM and IgG transcripts were amplified using polymerase chain reactions. Amplicons were sequenced using the Nanopore Native barcoding kit 24 V14 and MinION with R10.4 flow cells. In colostrum, there was a significantly greater percentage of IgM B cells with ultralong CDR3s (8.09% ± 1.73 standard error of the mean) compared to blood (4.22% ± 0.70, p = 0.05). There was a significantly greater percentage of IgG B cells in colostrum with ultralong CDR3s (12.98% ± 1.98) compared to blood (6.61% ± 1.11, p = 0.05). A higher percentage of IgM and IgG B cells with ultralong CDR3s in colostrum may be indicative of a potential role in protecting the neonate.

Novel electrochemical sensor based on poly([Cu(H2O)2P2]I2)/GCE for the determination of trimethoprim in clinical samples and cow's milk samples

A Novel bioanalytical detector based on Poly(Diaquabis(1,10-phenanthroline copper (II) Iodide) (poly([Cu(H2O)2P2]I2)/GCE) has been fabricated by potentiodynamic polymerization of [Cu(H2O)2P2]I2. The synthesized complex and the fabricated electrodes showed a promising electrocatalytic behavior towards the electrooxidation of Trimethoprim (TPM). Based on the regression coefficient value of scan rate with peak current and square root of peak current, the oxidation of TMP at poly([Cu(H2O)2P2]I2)/GCE was dominated by diffusion. Under the optimized experimental conditions and square wave voltammetric parameters, the poly([Cu(H2O)2P2]I2)/GCE revealed very wide responses in the range of concentration of 500 nM–160 μM with a limit of detection of 3.91 nM and a Limit of quantification of 13.02 nM with the relative standard deviation below 1.75 %. The valuable applicability of the fabricated sensor for the determination of TMP in real samples, including cow's milk, blood serum, and human urine, was successfully investigated. The electroanalysis results from the spike recovery and interference study provided an excellent range of recovery with an error percent of below 3.0 % in the combination effect with the potential interferents. The sensor exhibited excellent reproducibility, sensitivity, long-term stability, high accuracy, and fast response. The developed bioanalytical sensor revealed the best application prospect for biomedicine and environmental monitoring activities.

Screening of mtr-miR156a from exosomes of dairy cow blood to milk and its regulatory effect on milk protein synthesis in BMECs

MicroRNA (miRNA) is a type of endogenous non-coding small RNA, which is abundant in living organisms. miRNAs play an important role in regulating gene expression and myriad cellular processes by binding to target messenger RNAs through complementary base pairing, and cross-species regulation mammalian cells by plant-derived xeno-miRNAs has been described. Here, we examined the miRNA species in two alfalfa (Medicago sativa, lucerne) cultivars commonly grown in Ningxia, China: cv. Zhongmu 1 and cv. Xinyan 52. Both cultivars have good salt and drought resistance. We found that the miRNA profiles were similar between the cultivars, with a slightly higher number of miRNAs present in the newer cv. Xinyan 52, which may contribute to its improved salt and drought tolerance. miRNAs were stable during drying, and some miRNAs were increased in dry versus fresh alfalfa, suggesting some miRNAs may be upregulated during drying. Alfalfa-derived miRNAs could be detected in exosomes from serum and whey collected from dairy cows, confirming the ability of the exogenous miRNAs (xeno-miRNAs) to enter the circulation and reach the mammary epithelium. In vitro studies confirmed that overexpression of mtr-miR156a could downregulate expression of Phosphatase 2 Regulatory Subunit B'gamma ( PPP2R5D) and Phosphoinositide-3-kinase Regulatory Subunit 2 (PIK3R2). Overexpression of mtr-miR156a also modulated PI3K–AKT–mTOR signaling as well as the casein content of milk produced by bovine mammary epithelial cells. Based on the known roles of PPP2R5D and PIK3R2 in regulating the PI3K–AKT–mTOR pathway as well as the effect of PI3K–AKT–mTOR on milk protein content, our findings implicate alfalfa-derived miR156a as a new cross-species regulator of milk quality in dairy cows.

Structure of subopopulations of Aireshire, Holstin and Jersey cattle in the Southern Regions of Russia by CSN2 (rs43703011), CSN3 (rs43703016) and microsatellite locuses (BM1818, BM1824, BM2113, ETH10, ETH225, ETH3, INRA023, SPS115, TGLA126, TGLA122, TGLA227, TGLA53)

During studies using molecular biological methods, the following loci were genotyped: BM1818, BM1824, BM2113, ETH10, ETH225, ETH3, INRA023, SPS115, TGLA126, TGLA122, TGLA227, TGLA53, CSN2 (rs43703011) and CSN3 (rs43703 016) 3000 cow blood samples Ayrshire, Holstein and Jersey breeds from the herds of the Krasnodar and Stavropol territories. On average, the frequency of occurrence of homozygous genotypes for 12 microsatellite loci in Jersey cattle was 0.38, in Holstein and Ayrshire cattle – 0.30 and 0.27, respectively; the highest allelic diversity was found in Holstein cattle (9.0 ± 1.7 alleles per locus). It should be noted that there are significant differences in the genetic profile of Ayrshire cattle. For Ayrshire animals, the following alleles turned out to be typical (but rare in Jersey and Holstein cattle): 270 locus BM1818, 137 locus BM2113, 213 locus ETH10, 119 locus ETH3, 142 locus ETH225, 87 locus TGLA227, 166 locus TGLA53. In the studied subpopulation of Jersey cattle, the frequency of occurrence of the A2A2 genotype of the CSN2 locus (rs43703011) (beta casein) was 0.68, and in the Ayrshire and Holstein groups it was only 0.25 and 0.31, respectively. The BB genotype of the CSN3 locus (rs43703016) (casein kappa) was found in the Jersey group of animals with a frequency of 0.76, and in Ayrshires and Holsteins – 0.04 and 0.15, respectively. The genotyped groups of Jersey, Ayrshire and Holstein animals are significant in volume; we assume that the identified patterns are typical for cattle of these breeds in the south of Russia. We believe that the data obtained can be used in the future in practical work on classifying certain groups of livestock as typical Holstein, Ayrshire or Jersey and when creating herds that produce milk of a certain type (for example, type A2 milk).

Concentration of kisspeptin, testosterone and cortisol in the blood of cows during the transit period with energy metabolism disorders

In the clinically healthy cows of the Holstein breed (n = 20), a hormonal background was studied during theperiod of deepness and after the edema. For this purpose, the concentration of kisspeptin, testosterone and cortisol was determined in serum cows in the blood cows. The animals conducted regular observation with the conduct of clinical and obstetric and gynecological studies. Depending on the concentration of glucose and beta-hydroxybutyric acid on the 5th day after calving, the cows were divided into two groups. The first group included 15 heads with a glucose concentration greater than 3 mmol/l and a beta-hydroxybutyric acid level below 1 mmol/l. In the second group, 5 heads had violations in energy exchange (glucose levels less than 3 mmol/l, beta-hydroxybutyric acid above 1 mmol/l). On the 15th day after the hotel, normalization of energy homeostasis was recorded. The concentration of kisspeptin in the preventive period in both groups decreased to the hotel. At the same time, the concentration of kisspeptin in the blood of animals in the group with energy exchange disorders was 2.4–3 times higher on the 9th, 7th and 5th days before calving (p &lt; 0.01–0.05) compared with the first group. In production experience, the concentration of all three hormones after the hotel decreased with significant intergroup differences. The level of kisspeptin was reliably higher in the preventive period in the experimental group and on the 5th day after the hotel (p &lt; 0.05). The cortisole level was held at a high level of up to 7 days after the hotel (p &lt; 0.05). The concentration of testosterone in the blood throughout the transit period was reliably high in the animals of the experimental group.

Association between IGF-1 and IGFBPs in Blood and Follicular Fluid in Dairy Cows Under Field Conditions.

Insulin-like growth factor 1 (IGF-1) regulates dairy cow reproduction, while the paracrine IGF system locally influences fertility. In both systems, IGF-1 bioactivity is regulated through binding proteins (IGFBPs) inhibiting IGF-1 binding to its receptor (IGF1R). This study aimed to investigate a possible transfer between this endocrine and paracrine system. Therefore, blood and follicular fluid (FF) from postpartum dairy cows were analysed for ß-hydroxybutyrate (BHB), IGF-1, IGFBP-2, -3, -4, -5, and an IGFBP fragment in two study parts. The mRNA expression of IGFBP-2, IGFBP-4, IGF1R, and the pregnancy-associated plasma protein A (PAPP-A) in granulosa cells was measured. The results showed correlations between plasma and FF for IGF-1 (r = 0.57, p < 0.001) and IGFBP-2 (r = -0.57, p < 0.05). Blood BHB negatively correlated with IGF-1 in blood and FF and IGFBP-3, -5 and total IGFBP in blood (IGF-1 plasma: r = -0.26, p < 0.05; FF: r = -0.35, p < 0.05; IGFBP-3: r = -0.64, p = 0.006; IGFBP-5: r = -0.49, p < 0.05; total IGFBP: r = -0.52, p < 0.05). A negative correlation was found between IGFBP-2 expression and IGF-1 concentration in FF (r = -0.97, p = 0.001), while an IGFBP fragment positively correlated with IGF1R-mRNA in FF (r = 0.82, p = 0.042). These findings suggest a transfer and local regulation between the somatotropic axis and the follicular IGF system, linking the metabolic status with local effects on dairy cow fertility.

The influence of probiotic administration on the phagocytic and oxidative burst activity of neutrophils and monocytes in the peripheral blood of dairy cows during different lactation periods

Abstract Introduction The objective of this bovine peripheral blood study was a comparative assessment of the phagocytic activity of neutrophils and monocytes and of the intracellular killing capacity of neutrophils from cows given no probiotic and from cows which were administered a probiotic consisting of Saccharomyces cerevisiae, Lactobacillus acidophilus, Lactobacillus plantarum and Rhodopseudomonas palustris. These activity types were compared during different lactation periods. Material and Methods A cohort of 20 pregnant dairy cows was divided into two groups of 10. The experimental group consisted of cows fed a ration supplemented with probiotics, and the control group consisted of cows fed an unsupplemented ration. Blood was drawn six times: 7 days before dry off, 14 days before parturition, and 7, 21, 60 and 90 days postpartum (DPP). The phagocytic activity of neutrophils and monocytes and the oxidative burst activity of neutrophils were determined by flow cytometry in the peripheral blood of all examined cows. Results Phagocytosis testing revealed increased percentages of phagocytic neutrophils and monocytes in the experimental group at 21, 60 and 90 DPP (P-value &lt; 0.01). The mean fluorescence intensity (MFI) values for neutrophils and monocytes were higher on all days of the study (P-value &lt; 0.01). In oxidative burst testing, the percentages of detected neutrophils and their MFI were increased in the experimental group on all days (P-value &lt; 0.01). Conclusion The use of probiotics supported dairy cows’ immunity throughout the whole experiment. Probiotic supplementation may limit the occurrence of infectious diseases in these animals.

Iodine deficiency as an environmental risk factor for thyroid gland diseases in animals

Among non-infectious diseases in humans and animals, iodine deficiency pathology is a significant problem worldwide due to its high prevalence and broad spectrum of clinical signs and consequences. Increasing anthropogenic pressure on the natural environment has led to an ecological crisis. The situation is complicated in the biogeochemical zones of Ukraine, where insufficient or excessive content of mobile forms of microelements in soils and water sources determines the biological reaction of local flora and fauna. In animals, this is manifested by endemic diseases, including hypothyroidism. Mineral elements, including iodine, are a structural material in the body of animals; they participate in the processes of digestion and absorption in the digestive tract, synthesis, splitting, and excretion of metabolic products, creating the necessary conditions for the normal functioning of enzymes, hormones, vitamins, stabilize acidic-alkaline balance and osmotic pressure. Of all the organs of the endocrine system, the thyroid gland is the most closely related to the environment since its structure and function largely depend on the supply of iodine from the outside. The authors describe in detail the role of synergists of iodine (selenium, iron, copper, zinc, cobalt) and antagonists, the imbalance of trace elements, the influence of ionizing radiation, and the nature of feeding. It is substantiated that the action of environmental elements and anthropogenic factors determines the appearance and risk of developing thyroid pathology in cows. The article presents materials from a complex study of the clinical course of iodine deficiency, as well as morphological and biochemical parameters of the blood of cows. The object of research is clinically healthy and sick animals – cattle. Diagnosis of thyroid pathology was carried out with the help of clinical and laboratory tests. The clinical examination of cows was carried out according to the generally accepted scheme, taking into account the results of the general condition and studies of individual organs and systems. The research was conducted in three districts of the Zhytomyr region (Korostenskyi, Narodytskyi, and Popilnianskyi). During the external examination of the animals, swelling in the intermaxillary space (Myxedema) was observed, which was found only in 5 out of 90 dairy cows (5.6 %), mainly from Narodytsky district (8.9 %), compared to 2.2 % in Korosten district. It was observed that goiter was established in only three cows out of 90 (3.35%); all were in Narodytsky district (6.7 %). Enophthalmos was detected in 24 out of 90 dairy cows (26.7 %), including 16 out of 45 cows (35.4 %) in the Narodytsky District, while this symptom was not detected in the Popilnya District dairy cows. It was established that the content of thyroxine in dairy cows of Narodytskyi and Korostensky districts ranged from 2.2 to 4.25 μg/100 ml (28.3–54.7 nmol/l), respectively, and was on average 3.4 ± 0.21 μg/100 ml (43.8 ± 2.70 nmol/l, in cows from the conditionally clean territory (Popelnyan district) – 5.3 ± 0.65 nmol/l.

Heavy metal impacts on antioxidants in cow blood from wastewater-irrigated areas

The aim of the present research was to investigate the presence of heavy metals such as lead (Pb), copper (Cu), chromium (Cr), and cadmium (Cd) in blood samples from cows raised with irrigated wastewater, as well as in the wastewater itself, in the North-western region of Pakistan. A total of 60 blood samples were collected from five different locations in Kohat, namely Tappi Road (TR), Pindi Road (PR), Gul Malik Road (GMR), Markaz Road (MR), and a control group. The samples of both i.e. cow blood and wastewater were analyzed for the concentrations of heavy metals. The highest concentration of Cd was detected in the MR site with a mean value of 0.03 mg/L, and the highest concentration of Cu (0.04 mg/L) was recorded in the TR site, while the lowest level was found in the control group with a mean of 0.002 mg/L in blood samples. The highest Cr and Pb concentrations were found at the PR site, with mean values of 0.03 and 0.07 mg/L, respectively, whereas the control group had the lowest concentrations, with mean values of 0.002 and 0.01 mg/L. Similarly, heavy metal concentrations were analyzed in wastewater used for irrigation in the study area. Results indicated elevated concentrations of Cu and Cr in wastewater, although they remained below the World Health Organization (WHO) recommended values except for Cr (0.13 mg/L) in the GMR site, which exceeded permissible limits. Cd and Pb concentrations in wastewater were relatively low, but Cd concentration surpassed WHO limits, particularly with a mean concentration of 0.08 mg/L in the TR site. Comparison between heavy metal concentrations in blood and wastewater revealed higher values of Cd and Pb in blood samples than in wastewater, while Cu and Cr concentrations were higher in water compared to blood. Additionally, elevated levels of Super Oxide Dismutase (SOD), antioxidant enzyme Catalase (CAT), and oxidative stress marker malondialdehyde (MDA) were detected in blood samples. Cluster and principal component analyses were employed to assess heavy metal toxicity among the groups, indicating potential long-term adverse health effects on animals, transfer to humans, and toxicity in living organisms.

Milk productivity and the concentration of ketones in the blood of Kostroma cows of different LEP genotypes

Relevance. Currently, scientists are searching for genes associated not only with an increased level of dairy productivity, but also with the preservation of functional longevity and health of cows, with a high level of realization of their genetic potential, as well as with the distinctive quality of dairy products. The available data allow us to consider the leptin gene (LEP) as one of the candidate genes of milk productivity.Methods. The research was conducted in 2022–2023 on cows of the Kostroma breed (n = 107) bred in the Kostroma region. In cows in the postpartum period, the content of ketone bodies in the blood and genotypes were determined by the R25C locus of the LEP gene. The animals were divided into groups depending on the concentration of ketone bodies in the blood: 1st – 0–1.2 mmol/l, 2nd – above 1.2 mmol/l.Results. The presence of a predisposition of highly productive cows to ketosis (р &lt; 0.05) was established. The largest number of ketone bodies in the blood was determined in animals of the 2nd group with the LEPCT genotype. However, despite such a small difference, the difference in the frequency distribution of LEP alleles in groups 1 and 2 turned out to be statistically significant (χ2 = 63.48, р &lt; 0.001). When testing the hypothesis of a significant dependence of the level of milk productivity on the concentration of ketone bodies in the blood of carriers of the CC genotype, a statistically significant rho value was obtained among cows with the CC genotype, at the level of 0.37 (р = 0.038). The rank correlation between milk yield and ketone body content in carriers of LEPCT and LEPTT genotypes was low and unreliable (р &gt; 0.05). The tendency towards a close relationship between the level of ketone bodies and the fat content in the milk of cows of the TT genotype due to its low concentration in the sample requires further confirmation in a larger sample of cows with this genotype of the leptin gene.

Effects of grain intervention on hypothalamic function and the metabolome of blood and milk in dairy cows

BackgroundThe hypothalamus plays a crucial role in the health and productivity of dairy cows, yet studies on its functionality and its impact on peripheral circulation in these animals are relatively scarce, particularly regarding dietary interventions. Therefore, our study undertook a comprehensive analysis, incorporating both metabolomics and transcriptomics, to explore the effects of a grain-based diet on the functionality of the hypothalamus, as well as on blood and milk in dairy cows.ResultsThe hypothalamic metabolome analysis revealed a significant reduction in prostaglandin E2 (PGE2) level as a prominent response to the grain-based diet introduction. Furthermore, the hypothalamic transcriptome profiling showed a notable upregulation in amino acid metabolism due to the grain-based diet. Conversely, the grain-based diet led to the downregulation of genes involved in the metabolic pathway from lecithin to PGE2, including phospholipase A2 (PLA2G4E, PLA2G2A, and PLA2G12B), cyclooxygenase-2 (COX2), and prostaglandin E synthase (PTGES). Additionally, the plasma metabolome analysis indicated a substantial decrease in the level of PGE2, along with a decline in adrenal steroid hormones (tetrahydrocortisol and pregnenolone) following the grain-based diet introduction. Analysis of the milk metabolome showed that the grain-based diet significantly increased uric acid level while notably decreasing PGE2 level. Importantly, PGE2 was identified as a critical metabolic marker in the hypothalamus, blood, and milk in response to grain intervention. Correlation analysis demonstrated a significant correlation among metabolic alterations in the hypothalamus, blood, and milk following the grain-based diet.ConclusionsOur findings suggest a potential link between hypothalamic changes and alterations in peripheral circulation resulting from the introduction of a grain-based diet.

Evaluation of a Commercial Pregnancy Test Using Blood or Plasma Samples in High-Producing Dairy Cows.

This study evaluated a commercial pregnancy-associated glycoproteins (PAGs)-based pregnancy test using whole blood or plasma samples during early pregnancy (28-55 days of gestation) in high-producing dairy cows. Transrectal ultrasonography was used as the gold standard method. The study population constituted of 284 cows. False positive diagnoses were recorded from Day 60 to 89 and from Day 60 to 99 postpartum in blood and plasma samples, respectively. In early pregnancy screening, correct positive diagnoses were recorded in 75% and 100% of blood and plasma samples, respectively. High milk production was associated with negative results in blood samples and with the lowest test line intensity in plasma samples. False positive or negative diagnoses were recorded in 0% of both types of samples in cows previously diagnosed as pregnant and showing signs of estrus. In conclusion, the use of plasma was more effective than the use of blood in early pregnancy diagnosis. In cows previously diagnosed as pregnant and showing signs of estrus, both types of samples showed the same results. Because of large individual variations, normal single pregnancies could not be differentiated from twin pregnancies, from pregnancies with a recently dead conceptus, or from pregnancies that experienced subsequent pregnancy loss.