Transition Diseases in Grazing Dairy Cows Are Related to Serum Cholesterol and Other Analytes

The objectives of this study were to describe the incidence of postpartum disease and to evaluate the association with serum cholesterol concentrations during the first 3 weeks after calving in grazing dairy cows. The association between non-esterified fatty acids (NEFA), β-hydroxybutyrate (BHBA), calcium and postpartum diseases was also evaluated. A total of 307 Holstein dairy cows from 6 commercial grazing herds in Osorno, Chile, were monitored from calving until 21 days in milk. Cases of retained placenta, clinical hypocalcemia and clinical mastitis were recorded by the farmer using established definitions. Twice weekly, cows were evaluated for metritis by the same veterinarian based on vaginal discharge and body temperature. Postpartum blood samples were collected weekly and analyzed for serum concentrations of cholesterol, NEFA, BHBA and calcium. Cows were considered as having subclinical ketosis if BHBA >1.2 mmol/L, and subclinical hypocalcemia if calcium <2.0 mmol/L in any of the 3 weekly samples. Overall, 56% of the cows studied developed at least one clinical or subclinical disease after calving. Incidence of individual diseases was 8.8% for retained placenta, 4.2% for clinical hypocalcemia, 11.7% for clinical mastitis, 41.1% for metritis, 19.9% for subclinical hypocalcemia and 16.6% for subclinical ketosis. Lower postpartum cholesterol in cows was associated with developing severe metritis or having more than one clinical disease after calving. For every 0.4 mmol/L decrease in serum cholesterol cows were nearly twice as likely to be diagnosed with multiple clinical diseases after calving. Higher BHBA concentrations and lower calcium concentrations during week 1 were associated with severe cases of metritis. Low serum calcium concentration during week 1 was also associated with developing more than one clinical disorder after calving. In conclusion, the incidence of postpartum diseases can be high even in grazing herds and lower serum cholesterol concentrations were associated with occurrence of clinical postpatum disorders.     

Evaluation of the Change of Serum Copper and Zinc Concentrations of Dairy Cows with Subclinical Ketosis

Ketosis in dairy cows can lead to poor reproductive success and decreased milk production. Since the serum concentrations of copper (Cu) and zinc (Zn) are closely associated with the health status of cows, we investigated whether serum concentrations of Cu and Zn differed in dairy cows with subclinical ketosis and healthy dairy cows. Blood samples of 19 healthy dairy cows and 15 subclinically ketotic dairy cows were collected from three farms, and the concentrations of β-hydroxybutyrate (BHBA), glucose, non-esterified fatty acids (NEFA), Cu, and Zn were determined. Subclinically ketotic dairy cows had significantly higher BHBA and NEFA levels (p?<?0.01) and lower glucose (p?<?0.01) than healthy dairy cows. Likewise, serum concentrations of Zn were significantly decreased (p?<?0.05) in dairy cows with subclinical ketosis. There was no significant difference observed for serum Cu concentration between healthy and subclinically ketotic dairy cows. This study suggests that a decreased serum Zn concentration could be a cause of decreased reproductive performance in subclinically ketotic dairy cows.     

Relationship between plasma beta-carotene concentrations during the peripartum period and ovulation in the first follicular wave postpartum in dairy cows

Beta-carotene functions independently of vitamin A in the reproductive performance of dairy cows. The concentrations of beta-carotene in plasma decrease during the dry period, and reach a nadir in about the first week postpartum. This coincides with a negative energy balance, which affects the onset of the first ovulation in early postpartum cows. Thus, we hypothesised that plasma beta-carotene concentrations during the peripartum period may affect ovulation in the first follicular wave postpartum in dairy cows. The aim of the present study was to investigate changes in the profiles of plasma beta-carotene concentrations during the peripartum period in ovulatory and anovulatory cows during the first follicular wave postpartum. We used 22 multiparous Holstein cows, which were fed a total mixed ration consisting of grass, corn silage and concentrate, and collected blood samples for beta-carotene and progesterone analysis from week 3 prepartum to week 3 postpartum when the period of day 0-6 after parturition was regarded as the parturient week (week 0). The first ovulation was confirmed using the profile of plasma progesterone concentrations and colour Doppler ultrasound. Thirteen cows ovulated during the first postpartum follicular wave. Parity, the dry-off period, calving interval, mastitis episodes, and actual 305 days' milk yield during the previous lactation, and milk composition in the last month during the previous lactation in this study did not differ between ovulatory and anovulatory cows. Differences in the plasma beta-carotene profile were observed between ovulatory and anovulatory cows. Plasma beta-carotene concentrations at week 3 prepartum were greater in ovulatory cows (2.97+/-0.24 mg/L) than in anovulatory cows (1.53+/-0.14 mg/L; P<0.001), after that its concentrations in ovulatory cows decreased and reached the lowest level at week 1 postpartum, although its concentrations in anovulatory cows remained unchanged. No differences in plasma beta-carotene concentrations between the two groups were observed postpartum. The present study indicates for the first time that the lower beta-carotene concentrations in plasma during the prepartum period is associated with anovulation during the first follicular wave postpartum.     

Effect of Organic Selenium Supplementation on Selenium Status,Oxidative Stress,and Antioxidant Status in Selenium-Adequate Dairy Cows During the Periparturient Period

The periparturient period represents a stressful time for dairy cows as they transition from late gestation to early lactation. Oxidation stress occurs during this period owing to the increased metabolic activity. Antioxidants supplementation slightly above the suggested requirements may be beneficial in relieving this kind of stress. The objective of this study was to determine whether supplementing selenium (Se) yeast to diets with adequate Se concentrations affects Se status, oxidative stress, and antioxidant status in dairy cows during the periparturient period. Twenty multiparous Holstein cows were randomly divided into two groups with ten replicates in each group. During the last 4 weeks before calving, cows were fed Se-yeast at 0 (control) or 0.3 mg Se/kg dry matter (Se-yeast supplementation), in addition to Na selenite at 0.3 mg Se/kg dry matter in their rations. The concentrations of Se, reactive oxygen species (ROS), hydrogen peroxide (H₂O₂), hydroxyl radical, malonaldehyde (MDA), α-tocopherol and glutathione (GSH), the activities of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) and catalase (CAT), and the total antioxidant capacity (T-AOC) in plasma or erythrocyte of dairy cows were measured at 21 and 7 days prepartum, and at 7 and 21 days postpartum. Cows fed Se-yeast supplement during the last 4 weeks of gestation had higher plasma Se and lower MDA concentrations at 7 days prepartum, and at 7 and 21 days postpartum, and had higher whole blood Se and lower plasma ROS and H₂O₂ concentrations at 7 and 21 days postpartum compared with control cows. Se-yeast supplementation increased plasma and erythrocyte GSH-Px activities and erythrocyte GSH concentration at 7 days postpartum as compared to Se-adequate control cows. Compared with control cows, the enhanced SOD and CAT activities, increased α-tocopherol and GSH concentrations, and improved T-AOC in plasma at 7 and 21 days postpartum in Se-yeast-supplemented cows were also observed in this study. The results indicate that feeding Se-adequate cows a Se-yeast supplement during late gestation increases plasma Se status, improves antioxidant function, and relieves effectively oxidative stress occurred in early lactation.     

Effects of a plant product consisting of green tea and curcuma extract on milk production and the expression of hepatic genes involved in endoplasmic stress response and inflammation in dairy cows

During the periparturient phase, cows are typically in an inflammation-like condition, and it has been proposed that inflammation associated with the induction of stress of the endoplasmic reticulum (ER) in the liver contributes to the development of fatty liver syndrome and ketosis. In the present study, the hypothesis that supplementation of dairy cows with a plant product consisting of green tea (95%) and curcuma extract (5%) rich in polyphenols attenuates inflammation and ER stress in the liver during early lactation was investigated. Twenty-seven cows were assigned to two groups, either a control group (n = 14) or a treatment group (n = 13). Both groups of cows received a total mixed ration, and the ration of the treatment group was supplemented with 0.175 g of the plant product per kg dry matter from week 3 prepartum to week 9 postpartum. Dry matter intake and energy balance during week 2 to week 9 postpartum were not different between the two groups. However, cows supplemented with the plant product had a greater amount of energy-corrected milk during week 2 to week 9 postpartum and lower concentrations of triacylglycerols and cholesterol in the liver in week 1 and week 3 postpartum than cows of the control group (p < 0.05). Cows supplemented with the plant product showed a trend towards a reduced mRNA concentration of haptoglobin (p < 0.10), while relative mRNA concentrations of eight genes of the unfolded protein response considered in the liver were not different between the two groups of cows. Relative hepatic mRNA concentration of fibroblast growth factor, a stress hormone induced by various stress conditions, was reduced at week 1 and week 3 postpartum in cows supplemented with the plant product (p < 0.05). Overall, the data of this study suggest that – although there were only minor effects on the occurrence of ER stress and inflammation – a supplementation of polyphenols might be useful to improve milk yield and prevent fatty liver syndrome in dairy cows.     

Inflammatory cytokine concentrations in uterine flush and serum samples from dairy cows with clinical or subclinical endometritis

The objective of this study was to compare the concentrations of inflammatory cytokines in uterine flush and serum from healthy postpartum dairy cows and cows with clinical or subclinical endometritis. Clinical endometritis was diagnosed by observation of vaginal discharges (>50% pus) and subclinical endometritis was diagnosed by evaluation of uterine cytology (neutrophils >18%) at 4 weeks postpartum. Uterine flush was obtained from 48 cows at 4, 6, and 8 weeks postpartum for evaluation of tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-6, IL-8, and IL-10 concentrations. Serum samples were obtained from 34 cows just after calving and at 1, 2, 4, 6, and 8 weeks postpartum for evaluation of TNF-α, IL-1β, and IL-6 concentrations. Concentrations of TNF-α, IL-6, and IL-10 were greater (P < 0.05) in cows with clinical endometritis than in cows with subclinical endometritis and healthy controls, whereas concentrations of IL-8 in both cows with clinical and subclinical endometritis were greater (P < 0.005) than in controls. Overall, IL-6 and IL-10 concentrations decreased during the postpartum period. IL-1β concentrations in cows with clinical endometritis decreased (P < 0.0005) during the postpartum, whereas concentrations in cows with subclinical endometritis and controls did not change significantly with time; at 4 weeks postpartum, concentrations were greater (P < 0.0001) in cows with clinical endometritis. There were no significant effects of group, sampling time, or interaction on serum cytokine concentrations. In conclusion, cows with endometritis have greater inflammatory cytokine concentrations in uterine flush than healthy cows, but no differences were observed in serum.     

Effects of an energy-dense diet and nicotinic acid supplementation on production and metabolic variables of primiparous or multiparous cows in periparturient period

It is well observed that feeding energy-dense diets in dairy cows during the dry period can cause metabolic imbalances after parturition. Especially dairy cows with high body condition score (BCS) and fed an energy-dense diet were prone to develop production diseases due to metabolic disturbances postpartum. An experiment was conducted to determine the effects of an energy-dense diet and nicotinic acid (NA) on production and metabolic variables of primiparous and multiparous cows in late pregnancy and early lactation which were not pre-selected for high BCS. Thirty-six multiparous and 20 primiparous German Holstein cows with equal body conditions were fed with energy-dense (60% concentrate/40% roughage mixture; HC group) or adequate (30% concentrate/70% roughage mixture; LC group) diets prepartum. After parturition, concentrate proportion was dropped to 30% for all HC and LC groups and was increased to 50% within 16 days for LC and within 24 days for HC cows. In addition, half of the cows per group received 24 g NA supplement per day and cow aimed to attenuate the lipid mobilisation postpartum. Feeding energy-dense diets to late-pregnant dairy cows elevated the dry matter (p < 0.001) and energy intake (p < 0.001) as well as the energy balance (p < 0.001) without affecting the BCS (p = 0.265) during this period. However, this did not result in any metabolic deviation postpartum as the effects of prepartum concentrate feeding were not carried over into postpartum period. Multiparous cows responded more profoundly to energy-dense feeding prepartum compared with primiparous cows, and parity-related differences in the transition from late pregnancy to lactation were obvious pre- and postpartum. The supplementation with 24 g NA did not reveal any effect on energy metabolism. This study clearly showed that energy-dense feeding prepartum did not result in metabolic imbalances postpartum in multiparous and primiparous cows not selected for high BCS. A genetic predisposition for an anabolic metabolic status as indicated by high BCS may be crucial for developing production diseases at the onset of lactation.     

Effect of Chromium Supplementation on Productive and Reproductive Performances and Some Metabolic Parameters in Late Gestation and Early Lactation of Dairy Cows

The objective of this study was to determine the effect of supplemental chromium as chromium-L-methionine (Cr-Met) on productive and reproductive performances and some metabolic parameters in late gestation and early lactation of dairy cows. Sixty multiparous Holstein dairy cows according to prior lactation, parity, body mass (682?±?33?kg), and expected calving date were divided equally (30 cows/treatment) and were randomly allocated to one of the two groups. One group received basal diet without Cr (control group) and another group received Cr-Met supplement added at manufacturer's recommended level (8?mg of "Cr"/head per day) from 21?days before expected calving date until 21?days of lactation. Supplemental Cr tended to increase milk yield (P?=?0.08) while percentage of lactose and lactose yield increased (P??0.05). Percentage of cyclic cows at 36?days postpartum and estrous behavior before AI was higher in the supplemented group.     

Peripartum changes in the bovine placenta related to fetal membrane retention

To understand fetal membrane retention in dairy cattle, we examined these tissues in the immediate peripartum period before tissue separation. Placentomes were collected at 270-280 d of gestation from pre-partum Holstein cows (n = 5) and at 1, 3, 6, and 12 h postpartum from a) cows releasing fetal membranes in less than 12 h (n = 7), b) cows retaining fetal membranes for more than 12 h (n = 5), and c) cows induced to calve with dexamethasone (releasing membranes in more than 12 h; n = 5). Subjective evaluations of necrosis, distribution, and condition of binucleate giant and of principal cells were made. Necrotic foci and binucleate giant cells were counted for each interval for which tissue was available. Necrosis, existing prepartum, was identical to that observed at 1 h postpartum in all treatment groups (P > 0.05). Necrosis gradually increased in all treatment groups with time (P < 0.05); thus necrosis is unrelated to initiation of calving and is indirectly related to fetal membrane retention. Many binucleate giant cells were observed prepartum. In cows releasing fetal membranes normally, a significant (P < 0.05) decline in these cell numbers had occurred by 1 h postpartum. When fetal membrane retention occurred, no such decrease in binucleate giant cells was observed before 12 h postpartum. Loss of binucleate giant cells appears necessary for, or must occur with, separation of fetal membranes.     

Variations in Copper Concentration and Ceruloplasmin Activity of Dairy Cows in Relation to Lactation Stages with Regard to Ceruloplasmin to Copper Ratios

Variations of copper (Cu) concentration and ceruloplasmin (Cp) activity in serum and plasma of dairy cows at different stages of lactation were assessed in 240 Holstein dairy cows. Furthermore, ceruloplasmin to copper ratios were also investigated. The cows were classified according to their lactation stages into four different groups as close-up stage (3–1 week(s) antepartum), fresh lactation stage (0–1 week postpartum), early lactation stage (3–5 weeks postpartum), and mid-lactation stage (15–18 weeks postpartum). Each group consisted of 60 multiparous cows. Serum and heparinised plasma samples were obtained from each cow. Concentrations of serum copper (sCu) and plasma copper (pCu) as well as activities of plasma Cp (pCp) were higher in the group of fresh lactation stage than other groups (P < 0.05). Serum Cp (sCp) exhibited no significant difference between fresh lactation and early lactation groups (P > 0.05). Low correlations were obtained between sCp and pCu, sCp and sCu, and sCu and pCu. Plasma copper concentration and plasma ceruloplasmin activity were higher than serum copper concentration and serum ceruloplasmin activity, respectively (P < 0.001). The ratios of Cp activity to Cu concentration (Cp/Cu) were not significantly changed in the different lactation stages of dairy cows (P > 0.05). Use of sCp/pCu and sCp/sCu rather than pCp/pCu will reduce the calculated value of Cp/Cu. Furthermore, for evaluation of copper status, use of sCp/sCu or sCp/pCu identified more animals as ‘low’ and ‘marginal’ than using pCp/pCu (P < 0.001). It can be concluded that ceruloplasmin and copper undergo a physiological increase just after calving; thus, their values should be interpreted with caution during assessment of copper status. Plasma measurements should be used for calculation of Cp/Cu, and further research is required to refine diagnosis criteria for use of such ratio in determining copper status in dairy cows.     

Variables associated with peripartum traits in dairy cows. I. Effect of dietary forages and disorders on voluntary intake of feed, body weight and milk yield

Daily voluntary intakes of feed by each of 89 Holstein cows were compared between day 220 of gestation and day 30 postpartum over a 21-month period. Diets designed to meet NRC requirements and which contained either chopped hay (29 cows), hay crop silage (HCS; 30 cows) or corn silage (CS; 30 cows) were compared prepartum (27 to 0 days), peripartum (1 day before to 3 days after calving) and postpartum (days 4 to 30 postpartum). Mixed rations, fed during lactation, were 60% forage and 40% concentrate dry matter (DM). Cow management was similar to commercial operations. The experimental hypothesis was that pre-disposition for partum and postpartum disorders (abnormal) could be related either to voluntary intake of different diets or to physical traits. Intakes (DM or estimated net energy) across diets decreased 30% between days 7 and 1 prepartum and averaged 18% and 20% lower peripartum and postpartum, respectively, in abnormal cows than in control cows. Seasonal effects on intakes were significant. In general, changes in body weight and condition and differences in udder edema and milk yield reflected intakes. The results support the original hypothesis.     

Prepartum and Postpartum Rumen Fluid Microbiomes: Characterization and Correlation with Production Traits in Dairy Cows

Microbes present in the rumen of dairy cows are essential for degradation of cellulosic and nonstructural carbohydrates of plant origin. The prepartum and postpartum diets of high-producing dairy cows are substantially different, but in what ways the rumen microbiome changes in response and how those changes may influence production traits are not well elucidated. Here, we sequenced the 16S and 18S rRNA genes using the MiSeq platform to characterize the prepartum and postpartum rumen fluid microbiomes in 115 high-producing dairy cows, including both primiparous and multiparous animals. Discriminant analysis identified differences between the microbiomes of prepartum and postpartum samples and between primiparous and multiparous cows. 18S rRNA sequencing revealed an overwhelming dominance of the protozoan class Litostomatea, with over 90% of the eukaryotic microbial population belonging to that group. Additionally, fungi were relatively more prevalent and Litostomatea relatively less prevalent in prepartum samples than in postpartum ones. The core rumen microbiome (common to all samples) consisted of 64 bacterial taxa, of which members of the genus Prevotella were the most prevalent. The Chao1 richness index was greater for prepartum multiparous cows than for postpartum multiparous cows. Multivariable models identified bacterial taxa associated with increased or reduced milk production, and general linear models revealed that a metagenomically based prediction of productivity is highly associated with production of actual milk and milk components. In conclusion, the structure of the rumen fluid microbiome shifts between the prepartum and first-week postpartum periods, and its profile within the context of this study could be used to accurately predict production traits.     

Variables associated with peripartum traits in dairy cows. III. Effect of diets and disorders on certain blood traits

BLood samples were collected from 89 Holstein cows on days 220 and 250 of gestation, within 24 hr prepartum and postpartum and on day 30 postpartum. Balanced diets which contained either chopped hay (29 cows), hay crop silage (HCS; 30 cows) or corn silage (CS; 30 cows) were fed from day 220 of gestation to day 30 postpartum. The purpose was to determine if variations in certain blood traits were indicative of peripartum and postpartum disorders. The blood traits evaluated were concentrations of plasma total protein, whole blood hemoglobin, packed cell volume, and white blood cells, and serum glutamic oxalacetic transaminase (SGOT), glucose, urea nitrogen, calcium, inorganic phosphorus, magnesium, potassium and sodium. No blood trait was useful to predict a disorder prior to its visual signs with one possible exception. Serum glucose and calcium were lower and SGOT and magnesium were higher peripartum which was prior to death of three cows from fat cow syndrome.     

Effects of body condition score on direct and indirect measurements of insulin sensitivity in periparturient dairy cows

Reductions in insulin sensitivity in periparturient dairy cows develop as a means to support lactation; however, excessive mobilization of fatty acids (FA) increases the risk for peripartal metabolic disorders. Our objectives were to investigate the effect of prepartum body condition score (BCS) on systemic glucose and insulin tolerance, and to compare direct and indirect measurements of insulin sensitivity in peripartal lean and overweight dairy cows. Fourteen multiparous Holstein cows were allocated into two groups according to their BCS at day −28 prepartum: lean (n = 7; BCS ≤ 3.0) or overweight; (n = 7; BCS ≥ 4.0). Liver biopsies were performed on day −27, −14 and 4, relative to expected parturition. Intravenous insulin or glucose tolerances tests were performed following each liver biopsy. Relative to lean cows, overweight cows exhibited lower dry matter intake, lost more BCS and displayed increased plasma FA and β-hydroxybutyrate concentrations and elevated liver lipid content during peripartum. Glucose clearance rate was lower for all cows postpartum. Prepartum BCS had minimal effects on insulin and glucose tolerance; however, the ability of the cow to restore blood glucose levels following an insulin challenge was suppressed by increased BCS. Glucose-dependent parameters of insulin and glucose tolerance were not correlated with surrogate indices of insulin sensitivity. We conclude that prepartum BCS had minimal effect on systemic insulin sensitivity following parturition. The observed inconsistency between surrogate indices of insulin sensitivity and direct measurements of insulin and glucose tolerance adds support to growing concerns regarding their usefulness as tools to estimate systemic insulin action in periparturient cows.     

Effects of dietary supplementation of pioglitazone on metabolism,milk yield,and reproductive performance in transition dairy cows

The objective of this study was to investigate the effect of dietary supplementation of pioglitazone (PGT), a specific ligand for PPARγ, on metabolic dynamics, milk production, and reproductive performance of transition dairy cows. Eighty multiparous Holstein cows in their second or more lactations were blocked by the calving date and parity and assigned randomly to four dietary groups (n = 20 cow/treatment) including control (no PGT−/−), supplemented with PGT (6-mg PGT/kg body weight) from Day −14 to +21 relative to parturition (PGT+/+) or only during prepartum (PGT+/−) or postpartum periods (PGT−/+). Postpartum body condition score and body weight loss decreased (P < 0.05) in all PGT-supplemented groups. Milk yield was not affected by PGT supplementation (P > 0.05). Percentage of milk fat decreased (P < 0.05) in all PGT-treated groups; however, milk fat yield was lower (P < 0.05) in PGT (+/+) and PGT (+/−) groups compared with PGT (−/−). Peripartum (Day −7 to +7) concentrations of plasma nonesterified fatty acids and β-Hydroxybutyrate decreased in PGT (+/+) but not in the PGT (−/−) group (P < 0.05). During the postpartum period, PGT reduced (P > 0.05) plasma concentrations of nonesterified fatty acids in all PGT-treated groups but did not affect β-Hydroxybutyrate level. Plasma concentrations of triglycerides decreased in all PGT-supplemented groups. Supplementation of PGT increased the peripartum concentrations of plasma glucose in PGT (+/+) and PGT (+/−) groups compared with control. Plasma concentrations of insulin-like growth factor 1 were higher in PGT (+/+) compared with the control group during both the peripartum and postpartum periods. Plasma concentrations of growth hormone and insulin were not affected by PGT treatment (P > 0.05). Mean days to ovulation were lower in PGT (+/+) and PGT (-/+), and the proportion of cows ovulating by Day 14 postpartum was higher in PGT (+/+) compared with control. Days open were shorter in PGT (+/+), PGT (+/−), and PGT (−/+) groups compared with control. However, the proportion of pregnant cows at 120 days in milk were higher in all PGT-supplemented groups. The results showed positive effects of dietary supplementation of PGT, especially supplementation during both the prepartum and postpartum periods, on metabolic dynamics, ovarian function, and reproductive performance in transition dairy cows.     

Infradian rhythm in hematology,cell mediated immunity and serum micronutrient concentration around peripartum periods in high yielding crossbred cows before and after micronutrient supplementation

Alteration in hematology and cell-mediated immunity around peripartum periods before and after micronutrient supplementation was studied in this investigation. Twelve (12) high-yielding cross-bred cows in advanced pregnancy were selected for the experiment and divided into two equal groups viz. supplemented (n = 6) and unsupplemented (n = 6). Supplemented animals were provided with a micronutrient mixture (@ 25 g per cow) as per the recommendation of NRC over the normal feeding. Unsupplemented group of cows were kept as control with standard farm feeding management. Blood samples (15 ml/cow) were collected from all the animals during days 30th, 15th, and 7th before calving, on the day of calving, and during days 7th, 15th, and 30th after calving and analyzed for hematology viz. hemoglobin (Hb), total erythrocyte counts (TEC), total leukocyte counts, and differential leukocyte counts. In vitro phagocytic index (PI) of neutrophils and lymphocyte proliferation response (LPR) were also evaluated. Copper (Cu), zinc (Zn), manganese, and selenium (Se) concentrations in serum were estimated by atomic absorption spectrophotometer. The Hb concentration was found to be significantly (p < 0.01) higher in supplemented group compared to the unsupplemented group of cows. There were no significant differences in Hb concentration during different days of peripartum in both the group of cows. There was a significantly (p < 0.001) higher TEC in supplemented group compared to unsupplemented group of cows, though it did not vary significantly between different days of peripartum in both the group of cows. There was a significantly (p < 0.001) higher PI in supplemented group compared to the unsupplemented group of cows. In both the groups, the PI of blood neutrophils was significantly (p < 0.001) lower during calving. The LPR did not differ significantly between the groups, but it differed significantly (p < 0.01) between different days of peripartum and was lowest on the day of calving in both the groups. The Cu, Zn, and SE concentration was found to be significantly (p < 0.01) higher in supplemented group compared to the unsupplemented group of cows, but remained unaltered throughout the peripartum period. The study indicated that there was a marked immune depression around peripartum (15 days precalving to 15 days postcalving) in terms of in vitro phagocytic activity of blood neutrophils and in vitro LPR. Supplementation of micronutrients improves the hematological status as well as immune status, so it was recommended to supplement these micronutrients for better productivity and disease resistance in high-yielding cross-bred cows around peripartum.     

Effects of elevated parameters of subclinical ketosis on the immune system of dairy cows: in vivo and in vitro results

Using an established model in which subclinical ketosis is induced, the response of differential blood counts and levels of various haematological variables, including the inflammatory marker haptoglobin (Hp), were tested over the last six weeks of parturition until the 56th day post-partum in cows with lower or higher body condition scores (LBC and HBC, respectively; n = 9/group). Animals in the HBC group evidenced subclinical ketosis whereas LBC animals were metabolically healthy. For in vitro examination with ß-hydroxybutyrate (BHB) as a further stimulus, peripheral blood mononuclear cell (PBMC) counts of cows with and without subclinical ketosis (n = 5/group) were observed. Counts of leucocytes, granulocytes and lymphocytes (LY) peaked at day 1 post-partum in HBC cows, with a more marked increase in heifers. In subclinical ketosis LY count increased again, with significantly higher values in the HBC group. The red blood cell (RBC) profile was affected by parity (counts were higher in heifers). Hp showed a positive linear correlation with BHB and non-esterified fatty acids (NEFA; R² = 0.41). PBMC from cows that were not pre-stressed with subclinical ketosis were more sensitive to increasing levels of BHB in vitro, as evidenced by both their higher proliferative capability and increased release of nitric oxide (NO). In summary, cows with subclinical ketosis showed a heightened immune response compared with metabolically healthy individuals, based on increased LY counts, increasing stimulative properties of PBMC and a relationship between Hp and typically increased values of BHB and NEFA. Concentrations of BHB in vivo during subclinical ketosis did not alter the proliferative capability of bovine PBMC in vitro, which was first significantly decreased at a dosage of 5 mM BHB.     

The effects of energy concentration in roughage and allowance of concentrates on performance,health and energy efficiency of pluriparous dairy cows during early lactation

The aim of this study was to investigate the effects of different energy supplies from roughage and concentrates on performance, health and energy efficiency during early lactation. For this purpose an experiment was conducted containing 64 pluriparous German Holstein cows from 3 weeks prepartum until 16 weeks postpartum. During dry period all cows received an equal dry cow ration. After calving, cows were assigned in a 2 × 2 factorial arrangement to one of four groups, receiving either a moderate (MR, 6.0 MJ NE_L) or a high (HR, 6.4 MJ NE_L) energy concentration in roughage and furthermore moderate (MC, 150 g/kg energy-corrected milk (ECM)) or high amounts of concentrates (HC, 250 g/kg ECM) on dry matter (DM) basis, which were allocated from an automatic feeding system. Higher allocation of concentrates resulted in an increase of DM intake at expense of roughage intake. HC cows had a higher milk yield than MC cows, whereas ECM was higher in HR cows due to a decrease of milk fat yield in MR groups. Energy balance and body condition score were elevated in HC cows, but no differences occurred in development of subclinical ketosis. Furthermore, energy efficiency variables were lower in HC groups because the greater energy intake was not associated with a considerable elevation of milk yield. Consistency of faeces did not indicate digestive disorders in any of the treatment groups although the faecal manure score was significantly lower in HR groups. Our results underline the importance of a high energy uptake from roughage, which can contribute to an adequate performance and beneficial efficiency, especially at lower amounts of concentrates in ration. Feeding concentrates on an average amount of 9.4 kg/d compared to 6.4 kg/d on DM basis improved the energy balance in our trial, but without consequences for metabolic blood variables and general health of the cows.     

Effect of Supranutritional Organic Selenium Supplementation on Postpartum Blood Micronutrients,Antioxidants, Metabolites,and Inflammation Biomarkers in Selenium-Replete Dairy Cows

Dairy cows have increased nutritional requirements for antioxidants postpartum. Supranutritional organic Se supplementation may be beneficial because selenoproteins are involved in regulating oxidative stress and inflammation. Our objective was to determine whether feeding Se-yeast above requirements to Se-replete dairy cows during late gestation affects blood micronutrients, antioxidants, metabolites, and inflammation biomarkers postpartum. During the last 8-weeks before calving, dairy cows at a commercial farm were fed either 0 (control) or 105 mg Se-yeast once weekly (supranutritional Se-yeast), in addition to Na selenite at 0.3 mg Se/kg dry matter in their rations. Concentrations of whole-blood (WB) Se and serum Se, erythrocyte glutathione (GSH), and serum albumin, cholesterol, α-tocopherol, haptoglobin, serum amyloid A (SAA), calcium, magnesium, phosphorus, non-esterified fatty acids, and β-hydroxybutyrate were measured directly after calving, at 48 h, and 14 days of lactation in 10 cows of each group. Supranutritional Se-yeast supplementation affected indicators of antioxidant status and inflammation. Cows fed a supranutritional Se-yeast supplement during the last 8-weeks of gestation had higher Se concentrations in WB (overall 52 % higher) and serum (overall 36 % higher) at all-time points, had higher SAA concentrations at 48 h (98 % higher), had higher erythrocyte GSH (38 % higher) and serum albumin concentrations (6.6 % higher) at 14 days, and had lower serum cholesterol concentrations and higher α-tocopherol/cholesterol ratios at calving and at 48 h compared with control cows. In conclusion, feeding Se-replete cows during late gestation a supranutritional Se-yeast supplement improves antioxidant status and immune responses after calving without negatively impacting other micronutrients and energy status.     

Postpartal ovarian activity of healthy cows and those affected by subclinical metabolic disorders

The postpartal ovarian activity was studied in healthy cows and in those affected by subclinical fatty liver disease and subclinical primary ketosis. Twenty-six dairy cows, 4–7 years old, with not less than 5000 l milk yield in the preceding lactation, were monitored for the following blood parameters: aspartate-amino-transferase (AST)/glutamicoxalacetic transaminase (GOT), albumin, total protein, total lipid, cholesterol, non-esterified fatty acid (NEFA) and triglyceride levels, glucose and ketones. The ratios albumin : total protein and NEFA : triglyceride were also calculated. Blood samples were collected once a week between days 255 and 260 of gestation and days 1–15 postpartum, and subsequently on the 30th, 45th and 60th days postpartum. Based on the biochemical parameters, the cows were assigned in retrospect to three groups: (1) healthy (n = 5), (2) subclinical fatty liver disease (n = 6), and (3) subclinical primary ketosis (n = 10). The postpartal ovarian activity was assessed by rectal palpation and by milk progesterone levels, determined by radioimmunoassay in samples collected 2–3 times weekly between days 5 and 70 postpartum.As early as 10–18 days postpartum, ovarian activity was evident in healthy cows. This was confirmed by the progesterone profiles indicating three regular ovarian cycles with normal hormone levels within 70 days postpartum. All five healthy cows were detected in estrus and only one failed to conceive after insemination.The six cows with subclinical fatty liver disease had no regular cycles and only a few small follicles were palpable on their ovaries. The milk progesterone fluctuated around the basal level (? 1.6 nmol/l). The first Graafian follicles appeared as late as days 30–35 postpartum and the luteal function commenced only between days 38 and 60, as indicated by palpation of corpora lutea and a concomitant rise in milk progesterone. The progesterone peaks were at the lower physiological limits characteristic of luteal function.The ovarian activity of the ten cows with subclinical primary ketosis was intermediate between that of healthy and fatty liver cows. The first follicles and corpora lutea were somewhat delayed, and the progesterone levels fluctuated considerably during the luteal phase. One cow in this group resumed ovarian inactivity during the 70-day postpartal observation period.