Effects of Brown Seaweed (Ascophyllum nodosum) Supplementation on Enteric Methane Emissions, Metabolic Status and Milk Composition in Peak-Lactating Holstein Cows.

The dairy industry contributes significantly to anthropogenic methane emissions, which have an impact on global warming. This study aimed to investigate the effects of a dietary inclusion of brown seaweed Ascophyllum nodosum on enteric methane emissions (EMEs), hematological and blood biochemical profiles, and milk composition in dairy cows. Eighteen Holstein cows were divided into three groups: CON (non-supplemented cows), BS50 (50 mL of 10% A. nodosum), and BS100 (100 mL of 10% A. nodosum). In each cow, measurements of EME, dry matter intake (DMI), and milk yield (MY), as well as blood and milk sampling with respective analyzes, were performed before supplementation (P1), after 15 (P2) days, and after 30 (P3) days of supplementation. A. nodosum reduced (p < 0.05) methane production, methane yield, and methane intensity in both BS50 and BS100, and raised DMI (p < 0.05) only in BS50. Total bilirubin (p < 0.05) was higher in BS50 compared to CON cows in P2, and triacylglycerols were lower (p < 0.05) in BS50 than in CON cows in P3. Higher milk fat content was found in BS50 than in CON cows in P3. C16:0 proportions were higher (p < 0.05) in BS50 and BS100 than in CON cows, while C18:3n-3 was higher (p < 0.05) in BS100 than in BS50 and CON cows in P3. Dietary treatment with A. nodosum reduced EMEs and showed the potential to increase DMI and to improve energy status as well as milk composition in peak-lactating dairy cows.

Dietary Supplementation of Chestnut Tannins in Prepartum Dairy Cows Improves Antioxidant Defense Mechanisms Interacting with Thyroid Status.

Cows in the peripartal period undergo changes in thyroid hormones and are susceptible to lipomobilization and/or oxidative stress. The addition of chestnut tannins as polyphenolic compounds in the diet may improve feed efficiency and prevent oxidative stress-related health disorders in transition cows. However, the relationship between chestnut tannin supplementation and thyroid function, which plays an important role in metabolic regulation, has not been investigated in dairy cows. This study was conducted to investigate the effects of chestnut tannin supplementation during the close-up period on thyroid status and to evaluate the interaction between thyroid hormones and oxidative stress biomarkers in prepartum dairy cows. Forty multiparous Holstein cows were fed either a diet containing chestnut tannins (CNTs, n = 20, 1.96 g chestnut tannins/kg feed, dry matter) or a non-supplemented diet (CON, n = 20) during the last 25 ± 2 days of gestation. Blood samples were collected on the first day of study (before chestnut tannin supplementation) and d 5 before parturition to measure hormonal and oxidative stress indices. Serum concentrations of T3 (p = 0.04) and T4 (p = 0.05) were higher in CNT cows than in the CON group on day 5 before parturition. Thyroid status of CNT cows was associated with higher serum total antioxidant capacity (T-AOC, p < 0.01), activities of superoxide dismutase (SOD, p = 0.03) and glutathione peroxidase (GPx, p = 0.01), and reduced glutathione concentration (GSH, p = 0.05). Serum thiobarbituric acid reactive substances (TBARS) were lower (p = 0.04) which was associated with lower aspartate aminotransferase (AST, p = 0.02), and lactate dehydrogenase (LDH, p = 0.01) activities in the CNT than in the CON group. Estradiol and progesterone did not differ between CNT and CON cows. Chestnut tannin supplementation improves antioxidant protection, prevents oxidation-reduction processes, reduces the degree of liver cell membrane damage, and protects thyroid tissue from damage, allowing higher T3 and T4 synthesis. Considering the importance of the thyroid hormone status before parturition, mechanisms of thyroid hormone regulation in CNT-supplemented dairy cows require more detailed investigations.

Obesity-related prepartal insulin resistance in dairy cows is associated with increased lipin 1 and decreased FATP 1 expression in skeletal muscle.

A number of alterations have been identified in lipid metabolism within adipose tissue and liver in obesity. Less is known about the capacity of skeletal muscle for the metabolism of fatty acids in obesity-related insulin resistance, though it is evident that dry cow muscles may contain increased triglyceride content. The current study was therefore undertaken to evaluate the skeletal muscle expression of proteins of the fatty acid metabolism in dry cows with different body condition scores (BCS). Sixteen Holstein-Friesian close-up cows were divided into 2 equal groups based on their BCS as optimal (3.25 ≤ BCS ≤ 3.5) and high (4.0 ≤ BCS ≤ 4.25). Blood samples collection and skeletal muscle biopsies were carried out at day 10 before calving. Blood serum was assayed for concentration of resistin using a bovine specific ELISA. Protein expression of insulin receptor beta subunit (IRß), glucose transporter 4 (GLUT4), fatty acid translocase (FAT/CD36), fatty acid transporter 1 (FATP1), carnitine palmitoyltransferase 1 (CPT1), AMP-acitvated protein kinase (AMPK) and lipin 1 were analyzed in semitendinosus muscle by immunoblot. Resistin differed non-significantly between high-BCS and optimal-BCS cows. Insulin-resistant lipid metabolism in obese cows was paralleled with increased skeletal muscle expression of lipin 1 and GLUT4, and decreased expression of IRß and FATP1. These data suggest that in obesity-related insulin resistance, metabolic capacity in dry cow skeletal muscles appears to be organized towards the synthesis of signaling intermediates rather than fatty acids oxidation and that altered fatty acid uptake does not contribute to this disposition.

Summer Season-Related Heat Stress Affects the Mineral Composition of Holstein Dams' Colostrum, and Neonatal Calves' Mineral Status and Hematological Profile.

This study examines the effect of season-related heat stress in dams on mineral status, and hematological profile of their calves. Ten Holstein dams (LowT group) were exposed to the 49-day-long period of low air temperature (5.2 ± 0.5 °C) and ten dams (HighT group) were exposed to a 53-day-long high air temperature (27.4 ± 0.4 °C). The dams' blood samples were taken one hour after parturition, while colostrum was sampled at 2, 14, and 26 h after calving. Ca, P, Mg, Na, K, Fe, Zn, Mn, and Se concentrations were significantly lower while Cu and Co concentrations were significantly higher at least in two examined colostrum sampling points in HighT group. Calf`s blood samples were taken before 1st colostrum intake, and on days 1, 2, 3, and 7 postnatal. Ca, P, Na, K, Fe, Cu, Zn, Mn, Se, and Co were significantly lower while concentration of Mg was significantly higher in calves that originated from HighT group at least at two examined sampling points. Season affected dam's blood mineral status but the pattern was not always the same as in calves, since Na, K, Cu, and Co were significantly higher in HighT group. Calves that originated from HighT group had significantly lower hemoglobin, hematocrit, MCV, MCH, and MPV values, as well as neutrophils and monocyte counts, while platelets, eosinophils, basophils, and lymphocyte counts were significantly higher. Season-related heat stress leads to a reduction of the most minerals in both dams' colostrum and calves' blood and certain minerals' deficiencies are linked to impaired calves' hematological adaptation.

Parathyroid hormone response in treatment of subclinical hypocalcemia in postpartum dairy cows.

The aim of this study was to evaluate different treatments of postpartum subclinical hypocalcemia in dairy cows with emphasis on PTH response. Twenty four cows with subclinical hypocalcemia (Ca 1.4-2.0 mmol/L), out of 180 Holstein cows tested, were assigned in one of three equal groups: (1) CON received no Ca supplementation, (2) PO-Ca received 1000 mL of oral Ca suspension, immediately after establishing hypocalcemia, and 500 mL of the suspension 12 h after, (3) IV-Ca intravenously received 500 mL of Ca immediately after establishing hypocalcemia. The blood samples were taken up to 48 h after initial treatment for determination of Ca, iP, Mg and PTH concentrations. Urine pH was also evaluated up to 48 h after initial treatment. Compered to CON cows that were hypocalcemic up to the 48 h, Ca concentrations were significantly higher in PO-Ca up to hour 8 and IV-Ca up to hour 24. At first 2 h, PTH concentrations were significantly lower, compared to CON, only in IV-Ca, while at hour 4 PTH concentrations were significantly lower in both treated groups. At hours 24 and 48, PTH concentrations were significantly lower only in PO-Ca compared to CON group. Urine pH values in CON indicate alkalosis and were significantly lower both in PO-Ca and IV-Ca group, up to 12 h and 24 h respectively. In conclusion, both treatments mitigated hypocalcemia in cows, but with significantly different PTH response. Due to adequate PTH response, normocalcemia is established in CON and expected transient hypocalcemia is avoided in IV-Ca cows.

Preventing ectoparasite infestations reduces glucocorticoid concentrations in the hair of cows - short communication.

The aim of this study was to compare hair corticosteroid concentrations in ectoparasiticide-treated (n = 10) and non-treated (n = 12) Holstein cows. Animals in the treated group received cyfluthrin three times, on days 0, 28 and 56 of the experiment. Hair samples were collected from all cows on days 0, 21, 42, 63, and 84 of the experiment for the determination of cortisol and corticosterone concentrations using immunoassays. The respective hair cortisol concentrations in the treated group averaged 209.78, 165.10, 260.78, 177.44 and 183.11 ng/g, while in the non-treated group hair cortisol concentrations averaged 172.58, 243.58, 309.73, 199.75 and 207.09 ng/g. These results indicate that the control of ectoparasites reduced hair cortisol levels in dairy cows. The respective hair corticosterone concentrations in the treated group averaged 19.06, 22.95, 21.95, 21.60 and 24.84 ng/g and in the non-treated group the hair corticosterone concentrations averaged 17.28, 21.94, 34.05, 26.27 and 26.91 ng/g. The results suggest that longterm stress can be estimated better by the determination of hair cortisol rather than corticosterone concentrations.

Serum Protein Electrophoretic Pattern in Neonatal Calves Treated with Clinoptilolite.

The objective of our study was to determine the effects of clinoptilolite supplemented in colostrum on the blood serum protein electrophoretic pattern of new-born calves. METHODS: Romanian Black and White new-born calves involved in the study were divided into 3 groups: the control group (C) that received colostrum without clinoptilolite, and experimental groups I (E1) and II (E2) that received colostrum supplemented with 0.5% and 2% clinoptilolite, respectively. The concentration of total protein and protein fractions (albumin, α1-globulin, α2-globulin, ß-globulin and γ-globulin) were analyzed by electrophoresis on cellulose acetate. RESULTS: At hour 30 after birth, concentrations of γ-globulins, ß-globulin and total protein in E1 group of calves were higher than in control group by 42.11% (p < 0.05), 28.48% (p > 0.05) and 18.52% (p > 0.05), respectively, and were higher, but not significantly, in group E2 compared to the control group. This was in accordance with a significant lower albumin/globulin ratio in groups E1 and E2 (29.35%, p < 0.05 and 35.87%, p < 0.05, respectively) than in control group at 30 h postpartum, which indicates an obvious increase of the globulins fraction in experimental groups. The conclusion: Clinoptilolite was effective in improving passive transfer in new-born calves, but it was more effective if added in colostrum with a dose of 0.5% than with a dose of 2%.

Influence of different seasons during late gestation on Holstein cows' colostrum and postnatal adaptive capability of their calves.

Season may affect calves' thermal comfort and behavior, but the data related to the overall influence of seasonal variations on dams' colostrum and postnatal adaptive capability of calves are limited. The aim of this study was to measure the effects of a 49-day-long low air temperature (LAT) season (5.20 ± 0.46 °C mean air temperature) and a 53-day-long high air temperature (HAT) season (27.40 ± 0.39 °C mean air temperature) on dams' colostrum quality and physiological, biochemical, hormonal, and oxidative stress parameters of their calves during the first 7 days of life. The dams' colostrum was sampled at 2, 14, and 26 h after calving, before feeding of their calves. Calves' blood samples were taken before the first colostrum intake and on days 1, 2, 3, and 7 of life. Calves' physiological parameters were measured on days 0 and 7. HAT season significantly reduced the quality of dams' colostrum. The ingestion of the low-quality colostrum, combined with the thermal discomfort during HAT season, probably provoked impaired physiological, biochemical, hormonal, and oxidative stress parameters in samples taken from the post-colostral calves. Additionally, intravenous glucose tolerance test was performed on day 7, which suggested an enhanced insulin response in HAT season calves. This study highlights the importance of adequate supporting strategies for the care of the late gestation cows and postnatal calves during the HAT season.

Effect of Peroral Administration of Chromium on Insulin Signaling Pathway in Skeletal Muscle Tissue of Holstein Calves.

The objective of this study was to investigate the effects of peroral administration of chromium-enriched yeast on glucose tolerance in Holstein calves, assessed by insulin signaling pathway molecule determination and intravenous glucose tolerance test (IVGTT). Twenty-four Holstein calves, aged 1 month, were chosen for the study and divided into two groups: the PoCr group (n = 12) that perorally received 0.04 mg of Cr/kg of body mass daily, for 70 days, and the NCr group (n = 12) that received no chromium supplementation. Skeletal tissue samples from each calf were obtained on day 0 and day 70 of the experiment. Chromium supplementation increased protein content of the insulin ß-subunit receptor, phosphorylation of insulin receptor substrate 1 at Tyrosine 632, phosphorylation of Akt at Serine 473, glucose transporter-4, and AMP-activated protein kinase in skeletal muscle tissue, while phosphorylation of insulin receptor substrate 1 at Serine 307 was not affected by chromium treatment. Results obtained during IVGTT, which was conducted on days 0, 30, 50, and 70, suggested an increased insulin sensitivity and, consequently, a better utilization of glucose in the PoCr group. Lower basal concentrations of glucose and insulin in the PoCr group on days 30 and 70 were also obtained. Our results indicate that chromium supplementation improves glucose utilization in calves by enhancing insulin intracellular signaling in the skeletal muscle tissue.

Obesity-driven prepartal hepatic lipid accumulation in dairy cows is associated with increased CD36 and SREBP-1 expression.

We investigated the hypothesis that obesity in dairy cows enhanced expression of proteins involved in hepatic fatty acid uptake and metabolism. Sixteen Holstein-Friesian close-up cows were divided into 2 equal groups based on their body condition score (BCS) as optimal (3.25≤BCS≤3.5) and high (4.0≤BCS≤4.25). Intravenous glucose tolerance test (GTT) and liver biopsies were carried out at day 10 before calving. Blood samples were collected before (basal) and after glucose infusion, and glucose, insulin and non-esterified fatty acid (NEFA) levels were determined at each sample point. In addition, ß-hydroxybutyrate and triglycerides levels were measured in the basal samples. The liver biopsies were analyzed for total lipid content and protein expression of insulin receptor beta (IRß), fatty acid translocase (FAT/CD36) and sterol regulatory element-binding protein-1 (SREBP-1). Basal glucose and insulin were higher in high-BCS cows, which coincided with higher circulating triglycerides and hepatic lipid content. Clearance rate and AUC for NEFA during GTT were higher in optimal-BCS cows. The development of insulin resistance and fatty liver in obese cows was paralleled by increased hepatic expression of the IRß, CD36 and SREBP-1. These results suggest that increased expression of hepatic CD36 and SREBP-1 is relevant in the obesity-driven lipid accumulation in the liver of dairy cows during late gestation.

A comparison of the concentrations of energy-balance-related variables in jugular and mammary vein blood of dairy cows with different milk yield.

The aim of this study was to compare the concentrations of blood variables obtained simultaneously from the jugular and mammary veins of dairy cows. Eighty Holstein cows were divided into four equal groups: dry, low- (LY), medium- (MY) and high-yielding (HY). Blood insulin, glucose, non-esterified fatty acid (NEFA), beta-hydroxybutyrate (BHBA) and urea concentrations were measured. The jugular and mammary vein (J/M) ratio between concentrations of each variable was calculated. Differences between the groups of cows in concentrations of variables in the jugular vein were not in accordance with those obtained for the mammary vein. J/M values for insulin and glucose concentrations were above 1.0 in all groups of cows. The ratios for NEFA and BHBA concentrations were under or equal to 1.0 in dry and LY cows but above 1.0 in the MY and HY groups, indicating that in MY and HY cows those metabolites are apparently utilised by the mammary gland. J/M values for urea were above 1.0 in dry and LY cows but less than 1.0 in groups MY and HY, indicating that in the latter case urea is apparently released by the mammary gland. In conclusion, J/M for NEFA, BHBA and urea may be useful for estimation of the critical point when the mammary gland receives insufficient energy precursors for its current activity.

Thyroid Hormones Concentrations during the Mid-Dry Period: An Early Indicator of Fatty Liver in Holstein-Friesian Dairy Cows.

Relationship between postpartal fatty liver and thyroid gland activity during the peripartal and mid dry periods was studied. Twenty one dry cows were chosen. Blood samples were obtained on days -30, -2, and +12 related to calving and analized for thyroxine (T(4)) and triiodothyronine (T(3)). A T(3)/T(4) ratio was calculated. Liver tissue samples were taken 12 d after calving and tested for the lipid content. Cows were divided into three groups: mild (<20% fat), moderate (20 to 30%), or severe fatty liver (>30%). Cows, that were affected with severe fatty liver, were hypothyroid prior to development of the condition due to lower T(4) concentrations, and had significantly lower concentration of T(3) and higher T(3)/T(4) ratios than cows with mild and moderate fatty liver. Thus, hypothyroid state during mid-dry period may be an early indicator of postpartal fatty liver and may provoke T(3)/T(4) ratio increase in this group of cows.

Effects of peroral insulin and glucose on circulating insulin-like growth factor-1, its binding proteins and thyroid hormones in neonatal calves.

There is disagreement in the literature about the ability of neonatal calves to absorb perorally administered insulin. This study evaluated the absorption of a bolus of insulin administered alone or with an energy souce and its effects on the circulating insulin-like growth factor system and thyroid hormones in newborn Holstein-Friesian calves. Within 1 h of dosing, mean serum insulin and triiodothyronine (T3) concentrations had increased considerably, whether the insulin was applied alone (n = 4) or together with glucose (n = 4), accompanied by marked hypoglycemia. No significant changes were observed in control calves (n = 4) given the vehicle solution. Increased serum glucose and T3 concentrations with no change in insulinemia occurred in a 4th group of calves given glucose alone. At 32 h of age and after 3 meals of colostrum there were no differences in glycemia, insulinemia, or proteinemia among the 4 groups of calves examined. Mean serum insulin-like growth factor-I (IGF-I) tended to decrease over this period in the control group. The decrease was more pronounced in the insulin-treated group but absent in both groups that received glucose. These differences were associated with equivalent differences in abundance of the 40-45K IGF-binding protein-3 (IGFBP-3); however, lower molecular mass IGFBPs were not affected. The results show that a pharmacological peroral dose of insulin can lead to rapid systemic alterations in the IGF/IGFBP system in neonatal calves that can be modified by simultaneous administration of a small energy supply in the form of glucose.