Metabolic changes in follicular fluid of the dominant follicle in high-yielding dairy cows early post partum

Characteristics of the intrafollicular environment to which the preovulatory oocyte is exposed may be one of the major factors determining subsequent fertility. The aim of our study was to examine to what extent metabolic changes that occur in early post partum high-yielding dairy cows are reflected in the follicular fluid (FF) of the dominant follicle (>8 mm). Nine blood samples were taken per cow from nine high-yielding dairy cows between 7 days before and 46 days after parturition. From Day 14 post partum on and together with blood sampling, FF samples of the largest follicle were collected from the same cows by means of transvaginal follicle aspiration. Serum and FF samples were analyzed using commercial clinical and photometric chemistry assays for glucose, beta-hydroxybutyrate (beta-OHB), urea, total protein (TP), triglycerides (TG), non-esterified fatty acids (NEFA) and total cholesterol (TC). All cows lost body condition during the experimental period (0.94+/-0.09 points) illustrating a negative energy balance during the experimental period. In FF, glucose concentrations were significantly higher and the TP, TG, NEFA and TC concentrations were significantly lower than in serum (P<0.05). The concentrations of glucose, beta-OHB, urea and TC in serum and in FF changed significantly over time (P<0.05). Throughout the study, changes of all metabolites in serum were reflected by similar changes in FF. Especially for glucose, beta-OHB and urea the correlations were remarkably high. The results from the present study confirm that the typical metabolic adaptations which can be found in serum of high-yielding dairy cows shortly post partum, are reflected in follicular fluid and, therefore, may affect the quality of both the oocyte and the granulosa cells.     

The consequences of metabolic changes in high-yielding dairy cows on oocyte and embryo quality

Unsatisfactory reproductive performance in dairy cows, such as reduced conception rates, in addition to an increased incidence of early embryonic mortality, is reported worldwide and has been associated with a period of negative energy balance (NEB) early post partum. Typically, NEB is associated with biochemical changes such as high non-esterified fatty acid (NEFA), high β-hydroxybutyrate (β-OHB) and low glucose concentrations. The concentrations of these and other metabolites in the follicular fluid (FF) of high-yielding dairy cows during NEB were determined and extensively analyzed, and then were replicated in in vitro maturation models to investigate their effect on oocyte quality. The results showed that typical metabolic changes during NEB are well reflected in the FF of the dominant follicle. However, the oocyte seems to be relatively isolated from extremely elevated NEFA or very low glucose concentrations in the blood. Nevertheless, the in vitro maturation models revealed that NEB-associated high NEFA and low glucose levels in the FF are indeed toxic to the oocyte, resulting in deficient oocyte maturation and developmental competence. Induced apoptosis and necrosis in the cumulus cells was particularly obvious. Furthermore, maturation in saturated free fatty acid-rich media had a carry-over effect on embryo quality, leading to reduced cryotolerance of day 7 embryos. Only β-OHB showed an additive toxic effect in moderately hypoglycemic maturation conditions. These in vitro maturation models, based on in vivo observations, suggest that a period of NEB may hamper the fertility of high-yielding dairy cows through increased NEFA and decreased glucose concentrations in the FF directly affecting oocyte quality. In addition to oocyte quality, these results also demonstrate that embryo quality is reduced following an NEB episode. This important observation may be linked to the typical diet provided to stimulate milk yield, or to physiological adaptations sustaining the high milk production. Research into this phenomenon is ongoing.     

Acute fasting in heifers as a model for assessing the relationship between plasma and follicular fluid NEFA concentrations

It is known from epidemiological studies that negative energy balance in early lactating dairy cows is related to a depression in reproductive performance. Elevated plasma concentrations of non-esterified fatty acids (NEFA) are a typical metabolic characteristic of these animals and are proposed as the possible link. The suggestion is that NEFA might have a direct effect on the ovary, by affecting development of the oocyte or the granulosa cells. However, no data is available concerning the relationship between the concentration of NEFA in follicular fluid and plasma. Therefore, a cross-over study with 10 heifers around 15 months of age was conducted to analyze this relationship and examine the suggested effects of the negative energy balance on follicular growth. Investigation of these effects was performed on fasted heifers. The experimental treatment consisted of feeding hay with a subsequent period of fasting, to induce elevated plasma NEFA concentrations. Sampling of follicular fluid was performed using transvaginal aspiration of follicles, which were standardized using a synchronization protocol. In addition, concentrations of glucose, insulin, NEFA, and estradiol were measured in plasma. Follicular estradiol and progesterone concentrations were also measured to assess the quality of the dominant follicle. Fasting resulted in significantly lower plasma glucose (P=0.0006) and plasma insulin (P<0.0001) concentrations, higher plasma estradiol (P=0.008) and higher NEFA (P<0.001) concentrations, and smaller follicles (P=0.04) with lower estradiol:progesterone (E/P) ratios (P=0.05). Concentrations of NEFA in follicular fluid and plasma were closely related. Given this close relationship, we concluded that the presence of high plasma NEFA concentrations might link energy metabolism in early lactation with fertility.     

The effect of feeding diets high in quickly degradable nitrogen on follicular development and embryo growth in lactating Holstein dairy cows

This study investigated the effect on follicular and embryo development of increasing quickly degradable nitrogen (QDN) intake in lactating Holstein dairy cows. Forty mature post-partum cows were fed one of two diets for a minimum of 10 weeks, starting 10 days before first insemination. The Control diet was a high production dairy ration. The High QDN diet comprised the Control ration plus 250 g urea/head/day. Both diets were formulated to ensure that the energy requirements of the cows were satisfied. The High QDN treatment resulted in a significant increase in milk urea, plasma urea and plasma ammonia concentrations. The highest plasma urea (8.2 mmol/l) and ammonia concentrations (120 micromol/l) were recorded within 7 days of the urea supplement being added to the diet. There was no effect of diet on plasma progesterone or glucose concentration. There was also no significant effect of treatment on follicular development or embryo growth. The results from this study suggest that the lactating cow can adapt to increased intakes of QDN if the increase starts at least 10 days before insemination.     

Lactation influences the serum level of leptin and growth hormone during the daily bathyphase in ewes

The aim of the study was to investigate the influence of early lactation on leptin and growth hormone (GH) during bathyphase. Forty lactating Sarda ewes were divided into two equal groups on the basis of their milk production levels: HIGH (>1100 g/day) and LOW (<900 g/day). From the 5th to the 110th day after lambing, every 15 days, body condition score (BCS) was recorded and milk samples were collected. At the same data point, blood sampling was performed and leptin, GH and, Non-Esterified Fatty Acids (NEFA) were assessed. On milk, fat and proteins were determined. Statistical differences were observed in BCS, leptin, GH, NEFA and fat concentration in milk between the two groups. BCS was lower in high group and decreased from the 20th to the 90th day of lactation. Leptin was higher in low group and increased from the 50th and the 65th day of lactation, in low and high groups, respectively. GH and NEFA were higher in high group and decreased from the mild lactation. In high group, BCS and milk yield were negatively correlated and leptin was negatively correlated with GH and NEFA. In low group, leptin was positively correlated with BCS and negatively correlated with the all studied parameters. GH and NEFA were positively correlated with both groups. In conclusion, milk yield had an effect on the leptin and GH plasma values recorded during their bathyphase.     

Comparison of oocyte developmental competence and follicular steroid content of nulliparous heifers and cows at different stages of lactation

Reduced reproductive performance and lower conception rates of lactating cows are closely associated with genetic progress for high milk production. In contrast, the fertility of nulliparous Holstein heifers has remained fairly stable over the years and appears to be markedly higher than that of mature lactating cows. Possible differences in oocyte quality and follicular steroid levels, which could be associated with the low fertility of high-lactating cows, were examined in 13-month-old heifers, cows around the time of first AI (60-95 d post-partum, yielding 49+/-2.4 kg/d) and cows at mid-lactation (120-225 d post-partum, yielding 37+/-2.1 kg/d). Estrus was synchronized by two doses of PGF2alpha and follicles (5-8 mm) were aspirated on days 4, 8, 11 and 15 of the cycle by an ultrasound-guided procedure. Oocytes were morphologically examined, matured in vitro, chemically activated and cultured for 8d. Cleavage rate and the proportion of developing parthenogenetic blastocysts were determined on days 3 and 8 post-activation, respectively. On day 17, heifers and cows received additional PGF2alpha and follicular fluids from preovulatory follicles were collected on day 19 perior to the expected estrus. Follicular-fluid volumes were similar in cows and heifers, as were estradiol, progesterone and androstenedione concentrations in the follicular fluid. Percentages of high-grade oocytes, proportions of cleaved oocytes and developed blastocysts did not differ between the groups. Results suggest that the fertility gap between nulliparous heifers and high-lactating cows is not directly related to steroid content in the preovulatory follicular fluid or oocyte developmental competence.     

Metabolic adaptation and reticuloruminal pH in periparturient dairy cows experiencing different lipolysis early postpartum

Metabolic adaptation includes an array of concerted metabolic and endocrine events that enable dairy cows bridging the period of energy deficit at the onset of lactation. The present study evaluated metabolic, endocrine and reticuloruminal pH changes in 30 (25 Holstein and five Simmental) periparturient dairy cows experiencing variable lipolysis early postpartum. Cows were fed the same close-up and fresh lactation diets and kept in the same management conditions. Blood samples were collected at day 14, and day 4, relative to expected parturition, and at day 2, and day 21 postpartum, and serum metabolites and hormones related to glucose and lipid metabolism, as well as concentrations of several liver enzymes and acute phase proteins were determined. Additionally, reticuloruminal pH was monitored every 10 min over the last 3 days of the observation period. BW and milk yields were recorded and balances of energy and protein were assessed. Based on serum concentration of non-esterified fatty acids (NEFA) postpartum, cows were retrospectively classified into low (n=8), medium (n=11), and high (n=11) lipolysis groups, with NEFA levels of <0.4 mmol/l, between 0.4 and 0.7 mmol/l, and >0.7 mmol/l, respectively. Overall, elevated NEFA concentrations in the High group went along with a higher ratio of NEFA to cholesterol and reduced insulin sensitivity. While serum glucose, energy deficit and BW loss did not differ, cows of the High group exhibited increased lactate concentrations in the serum, compared with the Medium group. No differences in liver enzymes and acute phase proteins were evidenced among fat mobilization groups, whereas concentration of serum billirubin was lowest in the Low group after parturition. Data of milk yield and milk energy output showed no differences among groups, despite divergences in calculated energy balance and BW change postpartum. Cows of the Low group tended to increase dry matter intake but also showed longer time duration of pH below 6.0 in the reticulorumen (on average 299 min/day compared with 99 and 91 min/day for Medium and High groups, respectively). Differences in metabolic, endocrine and reticuloruminal pH responses indicate diverse metabolic adaptation strategies of early-lactation cows to cope with energy deficit postpartum.     

Plasma estrone sulfate,clinical biochemistry,and milk yield of dairy cows carrying a fetus from a bull or its clone

The aim of this article was to compare plasma estrone sulfate (E₁SO₄), clinical biochemistry, and milk yield of dairy cows carrying a female fetus from a bull (BULL) or from its clone (CLONE), evaluating also the relationship between the former variables and the birth weight of the newborn. Sixteen recipient dairy Friesian heifers (10 BULL and 7 CLONE) received a female embryo, obtained by in vitro embryo production and sexing by polymerase chain reaction with the semen of the BULL or the CLONE. Blood samples on all cows were obtained before feed distribution in the morning from jugular vein from 4 weeks before to 4 weeks after calving, to be analyzed for metabolic profile. The samples from late gestation were also analyzed for E₁SO₄ concentration. To separately assess the effect of calf birth weight (CBW), data were categorized as follows: low (<39 kg; BWT-A), mid (39–46 kg; BWT-B), and high (>46 kg; BWT-C). The plasma concentrations of β-hydroxybutyric acid (BHB, P = 0.019), Na (P = 0.002), Cl (P = 0.026), strong cation–anion balance (P = 0.020), total bilirubin (P = 0.054), and α₁-globulin (P = 0.044) were higher in prepartum BULL recipients than those in CLONE, whereas BHB (P = 0.021) and Mg (P = 0.090) were higher in postpartum BULL recipients, while no differences were recorded in the remaining postpartum parameters. The CBW class had significant interaction with week of gestation on antepartum plasma estrone sulfate (P = 0.021), whereas CBW per se affected antepartum plasma BHB (P = 0.021), and nonesterified fatty acids (NEFA; P = 0.011) being higher in BWT-C which also had the lower NEFA concentration during postpartum. Milk yield was unaffected by the sire used, both for quantitative and qualitative aspects. Cows carrying heavier fetus (BWT-C) had a different lactation affected by month compared with the other 2 CBW groups. From these results, there were no differences between BULL and CLONE recipients. Estrone sulfate, BHB, and NEFA may be used to predict CBW and provide different nutritional management during gestation.     

Effects of live weight adjusted feeding strategy on plasma indicators of energy balance in Holstein cows managed for extended lactation

In early lactation, most of the dairy cows are in negative energy balance; the extent and duration depend in part on the feeding strategy. Previous studies showed an increased lactation milk yield by use of a live weight (LW) adjusted feeding strategy with a high energy diet before and a reduced energy diet after LW nadir compared with a standard diet throughout lactation. The objective of the present study was to examine how such an individualized feeding strategy affects plasma indicators of energy status. It was hypothesized that an energy-enriched diet until LW nadir will reduce the severity of the negative energy balance, and that the reduction in diet energy concentration from LW nadir will extend the negative energy balance period further. Sixty-two Holstein cows (30% first parity) were managed for 16 months extended lactation and randomly allocated to one of two feeding strategies at calving. Two partially mixed rations were used, one with a high energy density (HD) and a 50 : 50 forage : concentrate ratio, and one with a lower energy density (LD, control diet) and a 60 : 40 forage : concentrate ratio. Half of the cows were offered the HD diet until they reached at least 42 days in milk and a LW gain⩾0 kg/day based on a 5-days LW average, and were then shifted to the LD diet (strategy HD-LD). The other half of the cows were offered the LD diet throughout lactation (control strategy LD-LD). Weekly blood samples were drawn for analysis of plasma metabolites and hormones. Before the shift in diet, the HD-LD cows had higher glucose and lower beta-hydroxybutyrate and non-esterified fatty acids (NEFA) concentrations than the LD-LD cows. After the shift until 36 weeks after calving, plasma NEFA was higher in HD-LD than LD-LD cows. Insulin and insulin-like growth factor-1 were not affected by the feeding strategy. To conclude, in early lactation, the energy-enriched diet reduced the negative energy balance. Plasma NEFA was higher in HD-LD than LD-LD cows from diet shift until 36 weeks after calving, indicating a carry-over effect of the early lactation HD diet to late lactation metabolism.     

The effect of feeding propylene glycol to dairy cows during the early postpartum period on follicular dynamics and on metabolic parameters related to fertility

Postpartum dairy cows (n=35) were used to determine the effects of feeding propylene glycol (PG) on metabolic variables related to ovarian function and on oocyte developmental competence. Starting on Day 7 postpartum, each animal received an oral dose (500 ml) of either PG or water once daily. Blood samples were collected on Days 5, 15, 25 and 35 pp to measure insulin, non-esterified fatty acids (NEFAs), beta-hydroxybutyrate (BHB) and glucose concentrations. Oocytes were recovered by ultrasound guided follicular aspiration starting on approximately Day 30 postpartum and submitted to in vitro fertilization. Ovarian follicular activity was examined daily by ultrasonography from Day 7 until ovulation or Days 35-40 postpartum. Animals receiving PG had elevated insulin concentrations over the subsequent 90 min following dosing (P<0.05) compared to control animals. Glucose concentrations followed a similar pattern. Irrespective of treatment, concentrations of NEFA declined significantly from Days 15 to 35 postpartum. Administration of PG resulted in a decrease in NEFA (P<0.001) and BHB (P<0.001) over the subsequent 90 min compared to control animals. Treatment with PG had no effect on follicular dynamics, mean days to emergence of the first cohort of follicles postpartum, or days to dominance and duration of dominance for any follicular wave recorded postpartum. There was also no difference in mean days to first ovulation or in size of the preovulatory follicle between treatments. Oocyte quality as measured by blastocyst development after IVF was not affected by treatment. These results suggest that administration of PG has the ability to positively alter the systemic concentrations of a number of metabolic variables which have been related to fertility. However, we did not observe an effect of PG treatment on follicular dynamics or the length of the postpartum interval. An effect on oocyte developmental competence remains to be proven.     

Relationship between body condition score loss and mRNA of genes related to fatty acid metabolism and the endocannabinoid system in adipose tissue of periparturient cows

The endocannabinoid system (ECS) controls feed intake and energy balance in nonruminants. Recent studies suggested that dietary management alters the expression of members of the ECS in the liver and endometrium of dairy cows. The aim of this study was to determine the relationship between body condition score (BCS) loss and the mRNA abundance of genes related to fatty acid metabolism and the ECS in the subcutaneous adipose tissue (AT) of dairy cows. The BCS was determined in multiparous (3.2 ± 0.5 lactations) Holstein cows at −21 and 42 days relative to calving (designated as d = 0). Cows were grouped into three categories according to BCS loss between both assessments as follows: (1) lost ≤0.25 unit (n = 8, low BCS loss (LBL)), (2) lost between 0.5 and 0.75 units (n = 8, moderate BCS loss (MBL)) and (3) lost ≥1 unit (n = 8, high BCS loss (HBL)). Concentrations of haptoglobin and non-esterified fatty acids (NEFAs) were determined in plasma. Real-time PCR was used to determine mRNA abundance of key genes related to fatty acid metabolism, inflammation and ECS in AT. Milk yield (kg/day) between week 2 and 6 post-calving was greater in the LBL group (49.4 ± 0.75) compared to MBL (47.9 ± 0.56) and HBL (47.4 ± 0.62) groups (P < 0.05). The overall mean plasma haptoglobin and NEFA concentrations were greater in MBL and HBL groups compared with the LBL group (P < 0.05). The mRNA abundance of TNF-α, Interleukin-6 (IL-6) and IL-1β was greatest at 21 and 42 days post-calving in HBL, intermediate in MBL and lowest in LBL groups, respectively. Cows in the HBL group had the greatest AT gene expression for carnitine palmitoyltransferase 1A, hormone sensitive lipase and adipose triglyceride lipase at 21 and 42 days post-calving (P < 0.05). Overall, mRNA abundance for very long chain acyl-CoA dehydrogenase and peroxisome proliferator-activated receptor gamma, which are related to NEFA oxidation, were greater in MBL and HBL groups compared to the LBL group at 42 days post-calving. However, mRNA abundance of fatty acid amide hydrolase was lower at 21 and 42 days post-calving in HBL cows than in LBL cows (P < 0.05). In summary, results showed a positive association between increased degree of BCS loss, inflammation and activation of the ECS network in AT of dairy cows. Findings suggest that the ECS might play an important role in fatty acid metabolism, development of inflammation and cow’s adaptation to onset of lactation.     

Metabolite and ionic composition of follicular fluid from different-sized follicles and their relationship to serum concentrations in dairy cows

Metabolic changes in blood serum may be reflected in the biochemical composition of follicular fluid and could indirectly influence oocyte quality. The purpose of this study was to examine the biochemical composition of follicular fluid harvested from different-sized follicles and its relationship with that of blood serum in dairy cattle. Following slaughter, blood samples were collected from dairy cows (n=30) and follicular fluid aspirated from three size classes of non-atretic follicles (<4 mm, 6–8 mm and >10 mm diameter). Samples remained independent between cows and between size classes within cows. Serum and follicular fluid samples were assayed using commercial clinical and photometric chemistry assays for ions (sodium, potassium and chloride) and metabolites (glucose, β-hydroxybutyrate (β-OHB), lactate, urea, total protein, triglycerides, non-esterified fatty acids (NEFA) and total cholesterol). Results showed that follicular fluid concentrations of glucose, β-OHB and total cholesterol increased from small to large follicles and decreased for potassium, chloride, lactate, urea and triglycerides. There was a significant concentration gradient for all variables between their levels in serum and follicular fluid (P<0.05). Significant correlations were observed for chloride (r=0.40), glucose (r=0.56), β-OHB (r=0.85), urea (r=0.95) and total protein (r=0.60) for all three follicle size classes and for triglycerides (r=0.43), NEFA (r=0.50) and total cholesterol (r=0.42) for large follicles (P<0.05). The results from the present study suggest that the oocyte and the granulosa cells of dairy cows grow and mature in a biochemical environment that changes from small to large follicles. Furthermore, the significant correlation between the composition of serum and follicular fluid for the above-mentioned metabolites suggests that metabolic changes in serum levels will be reflected in the follicular fluid and, therefore, may affect the quality of both the oocyte and the granulosa cells.     

Effect of butaphosphan and cyanocobalamin on postpartum metabolism and milk production in dairy cows

The aim of this study was to determine the effect of butaphosphan and cyanocobalamin (BTPC) supplementation on plasma metabolites and milk production in postpartum dairy cows. A total of fifty-two Holstein cows were randomly assigned to receive either: (1) 10 ml of saline (NaCl 0.9%, control group); (2) 1000 mg of butaphosphan and 0.5 mg of cyanocobalamin (BTPC1 group); and (3) 2000 mg of butaphosphan and 1.0 mg of cyanocobalamin (BTPC2 group). All cows received injections every 5 days from calving to 20 days in milk (DIM). Blood samples were collected every 15 days from calving until 75 DIM to determine serum concentration of glucose, non-esterified fatty acids (NEFA), β-hydroxybutyrate (BHB), cholesterol, urea, calcium (Ca), phosphorus (P), magnesium (Mg), aminotransferase aspartate (AST) and γ-glutamyltransferase (GGT). The body condition score (BCS) and milk production were evaluated from calving until 90 DIM. Increasing doses of BTPC caused a linear reduction in plasma concentrations of NEFA and cholesterol. Supplementation of BTPC also reduced concentrations of BHB but it did not differ between the two treatment doses. Milk yield and milk protein had a linear increase with increasing doses of BTPC. A quadratic effect was detected for milk fat and total milk solids according to treatment dose, and BTPC1 had the lowest mean values. Concentrations of glucose, urea, P, Mg, AST, GGT, milk lactose and BCS were not affected by treatment. These results indicate that injections of BTPC during the early postpartum period can reduce NEFA and BHB concentrations and increase milk production in Holstein cows.     

Effect of chromium methionine supplementation on lactation performance,hepatic respiratory rate and anti-oxidative capacity in early-lactating dairy cows

Chromium may regulate dairy cow metabolism; a chelated formation of chromium methionine (Cr-Met) is available to the feed industry. The objective of this study was to investigate the effect of Cr-Met supplementation on lactation performance, hepatic respiratory rate and anti-oxidative capacity in early-lactating Holstein dairy cows. 64 multiparous cows were assigned to 16 blocks based on parity and milk yield and then the four cows in a block were randomly allocated to four treatment groups with 0, 4, 8 or 16 g/d of Cr-Met per cow supplemented to a basal diet. Cows were moved from an open dry lot to a naturally ventilated tie stall barn 2 weeks before treatment to adapt to this facility, fed and milked at 0630, 1400, and 1930 h every day. The experiment lasted for 12 weeks. Milk yield and composition were recorded weekly. Dry matter intake was measured every 2 weeks for a total of six times throughout the trial. The plasma variables were measured in weeks 4, 8 and 12 of the experiment. Supplementation of Cr-Met did not affect DM intake of cows. As the supplementation of Cr-Met increased, yields of milk, fat, energy corrected milk (P < 0.01) and lactose (P = 0.01) increased in a linear manner. In terms of plasma variables, insulin concentration decreased in a linear manner with Cr-Met supplementation. As for variables relating to hepatic respiration rate, concentrations of pyruvate and NAD in the plasma were increased in quadratic manners, and lactic dehydrogenase activity was linearly increased as Cr-Met feeding levels increased. Moreover, plasma glutathione peroxidase and superoxide dismutase activity were increased in a linear manner. In conclusion, our study suggested that Cr-Met supplementation improved lactation performance of early-lactating dairy cows through enhancing antioxidant capacity and hepatic cellular respiration.     

Effects of Dexamethasone and Insulin Alone or in Combination on Energy and Protein Metabolism Indicators and Milk Production in Dairy Cows in Early Lactation – A Randomized Controlled Trial

Objectives

This study investigated the effects of dexamethasone and insulin, when administered at 3^rd or 10^th day of lactation on energy and protein metabolism in dairy cows.

Materials and Methods

Two hundred Holstein cows were enrolled in a randomized controlled clinical trial. The cows were randomly assigned to receive 1 of 4 treatments at 3 or 10 days in milk: control group, 10-mL i.m. injection of sterile water, group insulin, s.c. injection of 100 units of insulin, group dexamethasone, i.m. injection of 20 mg of dexamethasone, group insulin plus dexamethasone, i.m. injection of 20 mg of dexamethasone and 100 units of insulin. The cows randomly assigned to receive the treatments on 3 or 10 days of lactation. Serum samples obtained at the time of enrollment, time of treatment and at 2, 4, 7 and 14 days after intervention. The sera were analyzed for β-hydroxybutyrate (BHBA), nonesterified fatty acids (NEFA), glucose, cholesterol, albumin, urea, and aspartate amino transferase (AST). Data were analyzed using a repeated measures mixed model that accounted for the effects of parity, body condition score, dystocia, retained placenta, metritis and the random effect of cow.

Results

There was no significant interaction of group of treatment and time of intervention (day 3 or 10 post-partum) on serum components. Cows that received insulin or dexamethasone alone or in combination, had lower BHBA 2 days after treatment compared with control cows, whereas concentrations of NEFA, were unaffected suggesting that glucocorticoids lipolytic effects do not appear to be important in healthy cows. AST activities significantly reduced in cows that received dexamethasone with or without insulin at 2 and 4 days after treatment. Albumin and urea concentrations 2 days after treatment were higher for cows that received dexamethasone only or dexamethasone plus insulin compared with control and Ins received cows. There were no treatment effects on test-day milk production, milk fat and protein percentages.

Conclusions

The results suggested that administration of glucocorticoids in early lactation resulted in short-term improvement of metabolism in postpartum dairy cows in biochemical terms.     

Effects of pituitary-derived bovine growth hormone on production parameters and biokinetics of key metabolites in lactating dairy cows at peak and mid-lactation

Changes in production parameters and metabolite biokinetics induced by treatment with pituitary-derived bovine growth hormone (bGH) were monitored at peak (c. 40 days) and mid-lactation (c. 130 days) in dairy cows. During treatment with bGH milk production increased by 6 and 14% at peak and mid-lactation respectively. At peak lactation the content of milk fat tended to increase, whereas milk protein tended to decrease and milk lactose decreased significantly. Yield of milk fat increased, but there was no change in the yield of milk protein and lactose. The content of milk fat tended to increase at mid-lactation. Milk protein decreased and there was no change in milk lactose. Yields of milk fat and lactose but not protein increased. Growth hormone exerted metabolic effects which differed with stage of lactation. At peak lactation plasma glucose concentration and its irreversible loss increased, plasma urea and acetate were unchanged and their irreversible losses tended to increase. No change was measured for plasma non-esterified fatty acids (NEFA) and the irreversible loss of NEFA decreased. At mid-lactation plasma concentrations of glucose and NEFA were increased, plasma urea decreased and acetate and 3-hydroxybutyrate tended to increase. Irreversible losses of NEFA increased, urea tended to decrease and acetate and glucose remained essentially constant. The results show that exogenous pituitary bGH exerts metabolic effects which result in the supply of increased nutrients to support milk synthesis. The metabolic effects differ with the stage of lactation, reflecting differences in physiological and/or nutritional state.     

Effect of blood plasma collected after adrenocorticotropic hormone administration during the preovulatory period in the sow on oocyte in vitro maturation

Reproduction may be affected by stressful events changing the female endocrine or metabolic profile. An altered environment during oocyte development could influence the delicate process of oocyte maturation. Here, the effect of simulated stress by media supplementation with blood plasma from sows after adrenocorticotropic hormone (ACTH) administration during the preovulatory period was assessed. Oocytes were matured for 46 hours in the presence of plasma from ACTH-treated sows, or plasma from NaCl-treated control sows, or medium without plasma (BSA group). The plasma used had been collected at 36 and 12 hours (±2 hours) before ovulation (for the first 24 hours + last 22 hours of maturation, respectively). Subsequent fertilization and embryo development were evaluated. Actin cytoskeleton and mitochondrial patterns were studied by confocal microscopy both in the oocytes and the resulting blastocysts. Nuclear maturation did not differ between treatments. Subtle differences were observed in the actin microfilaments in oocytes; however, mitochondrial patterns were associated with the treatment (P < 0.001). These differences in mitochondrial patterns were not reflected by in vitro outcomes, which were similar in all groups. In conclusion, an altered hormonal environment provided by a brief exposure to plasma from ACTH-treated sows during in vitro oocyte maturation could induce alterations in actin cytoskeleton and mitochondrial patterns in oocytes. However, these changes might not hamper the subsequent in vitro embryo development.     

Improved developmental ability of porcine oocytes grown in nude mice after fusion with cytoplasmic fragments prepared by centrifugation: A model for utilization of primordial oocytes

Primordial oocytes are a potential resource for medical and zoological application, but those of large animals have not yet been reported to show efficient embryonic development. In the present study, we established a pig model for production of blastocysts from primordial oocytes that had been grafted into nude mice and matured in vitro, in combination with fusion of cytoplasmic fragments. Neonatal porcine ovaries in which most follicles are at the primordial stage were minced and grafted into nude mice (Crlj:CD1-Foxn1^nu). About 60 days after detection of vaginal opening, the mice were given 62.5 U/mL porcine FSH for 2 weeks by infusion to enhance follicular development. Developmentally competent oocytes collected from porcine ovaries (conventional oocytes) were matured in vitro and subjected to serial centrifugation to prepare cytoplasmic fragments without a metaphase plate (cytoplasts). Three cytoplasts were fused by electrostimulation to an oocyte retrieved from a host mouse (xenogeneic oocyte) and matured in vitro. Then these fused oocytes were fertilized and subsequently cultured in vitro. No blastocysts were generated from xenogeneic oocytes without fusion of cytoplasm. When xenogeneic oocytes had been fused with three cytoplasts, the blastocyst rate increased significantly to 14.3%, comparable to that for untreated conventional oocytes (20.0%). The numbers of cells in blastocysts for these fused oocytes (37.2 cells/blastocyst) were not significantly different from those for conventional oocytes (25.4 cells/blastocyst). Our findings show that it is possible to use primordial oocytes of large mammals in combination with xenografting of ovarian tissue and also ooplasmic fusion.     

Impact of lameness on gait characteristics and lying behaviour of zero grazed dairy cattle in early lactation

The present study examines the impact of lameness on the time budgets and gait of dairy cattle during early lactation. Automated assessment of activity together with an objective method (using video motion analysis) of assessing the gait of the cattle was utilised.Twenty-five Holstein dairy cows were recruited to the study and were assessed during weeks 1, 6 and 12 of lactation. Lying behaviour was measured using IceTag? activity monitors which were attached to the right hind leg of the cow for 4 consecutive days during each study week. Cows were locomotion scored to evaluate the influence of lameness on gait and behaviour of cows. Cows that were lame in a hind limb had significantly shorter fore and hind stride lengths. They also tended (p = 0.06) to have a negative tracking distance and walk at a slower (p = 0.002) speed compared with cows that were considered to be non-lame. Lame cows spent 2 h more time lying down per day in comparison to non-lame cows. Cows spent significantly less time lying down during week 6 of lactation and more time standing in comparison to the cows in week 12. The lying behaviour of cows was not different during weeks 1 and 6 or weeks 1 and 12. Cows were significantly more active during week 1 than week 12 of lactation. However the activity of the cows during week 6 was not different from weeks 1 and 12 of lactation. The present study demonstrates that lameness influences stride characteristics and lying behaviour of zero grazed dairy cows.