ABSTRACT
Dairy cows consume inadequate amounts of feed in early lactation and during conditions and diseases such as excessive fatness, heat stress, and infectious diseases. Affected cows often experience increases in plasma concentrations of acute phase proteins consistent with the negative effect of inflammation on appetite. The acute phase protein orosomucoid 1 (ORM1), also known as alpha-1-acid glycoprotein, was recently reported to reduce appetite in the mouse through its ability to bind the full-length leptin receptor (Ob-Rb) and activate appetite-suppressing signal transducer and activator of transcription 3 (STAT3) signaling. These observations raise the possibility that ORM1 exerts appetite-suppressing effects in dairy cattle during periods of increased inflammatory tone. The applicability of this model was assessed in 2 ways. First, we asked whether ORM1 is regulated during periods of inadequate appetite such as the transition from late pregnancy to early lactation and periods of increased inflammatory tone. Plasma ORM1 was invariant in late pregnancy but increased 2.5-fold between parturition and d 7 of lactation. Gene expression studies showed that liver was the major source of this elevation with little contribution by adipose tissue or mammary gland. Additional studies showed that plasma ORM1 was not increased further by excessive fatness or by reproductive dysfunction in early lactation and was completely unresponsive to inflammatory stimuli such as heat stress or intravascular administration of the endotoxin lipopolysaccharide during established lactation. Second, we tested the ability of ORM1 to trigger STAT3 signaling through Ob-Rb using Chinese hamster ovary K1 (CHO-K1) cells transfected with a STAT3 expression plasmid. In this configuration, CHO-K1 cells did not express Ob-Rb and were incapable of leptin-induced STAT3 phosphorylation. Leptin responsiveness was conferred by co-transfecting with bovine Ob-Rb, with leptin causing increases of 5.7-fold in STAT3 phosphorylation and 2.1-fold in the expression of the STAT3-dependent gene, SOCS3. In contrast, neither bovine or human ORM1 triggered STAT3 phosphorylation irrespective of dose and period of incubation tested. In summary, bovine ORM1 is not increased during periods of increased inflammatory tone except in early lactation and is incapable of Ob-Rb-dependent STAT3 signaling. Overall, these data are inconsistent with ORM1 mediating the appetite-suppressing effects of inflammation in cattle through Ob-Rb.
Subject(s)
Appetite Regulation/physiology , Cattle/metabolism , Lactation/physiology , Orosomucoid/metabolism , Receptors, Leptin/metabolism , STAT3 Transcription Factor/metabolism , Signal Transduction , Acute-Phase Proteins/metabolism , Adipose Tissue/metabolism , Animals , CHO Cells , Cricetinae , Cricetulus , Female , Leptin/metabolism , Pregnancy , Up-RegulationABSTRACT
Most dairy cows develop a dominant follicle within two weeks postpartum, but 60% of these follicles fail to ovulate. In a previous study, we determined that cows destined to ovulate have higher LH pulse frequency and circulating estradiol. The latter characteristic provided a method for distinguishing ovulatory from nonovulatory follicles during development and we found that nonovulatory follicles have lower estradiol and androstenedione in their follicular fluid. We hypothesized that lower LH pulse frequency impairs androgen production by theca cells of nonovulatory cows, reducing their ability to make estradiol. In the present study, we applied our method for predicting follicle fate to collect dominant follicles from predicted ovulatory (n = 7) and nonovulatory (n = 3) follicles. Theca and granulosa cells were separated and cultured in the absence or presence of LH, FSH, and/or testosterone for three days, with daily collection of culture medium for steroid RIAs. Estradiol and progesterone production by granulosa cells were not different between ovulatory and nonovulatory follicles. By contrast, overall androstenedione production by theca cells from ovulatory follicles was significantly higher compared with nonovulatory follicles on all three days of culture and, as culture progressed, theca from nonovulatory follicles had increasingly poorer responses to LH. In the same cultures, the progesterone production by theca cells was similar in ovulatory and nonovulatory groups. In support of our hypothesis, the results show that estradiol production by granulosa cells from nonovulatory follicles is robust when androgen substrate is present, but that thecal androgen production in response to LH is impaired. This suggests that the initial defect in steroidogenesis in dominant follicles that fail to ovulate postpartum is lower production of androgen by theca cells.
Subject(s)
Androgens/metabolism , Androgens/pharmacology , Cattle/physiology , Luteinizing Hormone/pharmacology , Theca Cells/drug effects , Theca Cells/metabolism , Animals , Cells, Cultured , Dose-Response Relationship, Drug , Estradiol/metabolism , Female , Granulosa Cells/metabolism , Luteinizing Hormone/administration & dosage , Postpartum Period , Pregnancy , Progesterone/metabolismABSTRACT
This study was conducted to evaluate associations of polymorphisms in the genes for the growth hormone receptor (GHR), insulin-like growth factor I (IGF-I) and signal transducer and activator of transcription 5A (STAT5A) with serum concentrations of IGF-I, reproductive performance and milk production of postpartum Holstein dairy cows. Days from calving to first ovulation (DTO) and calving to conception interval (CCI) were evaluated in 95 Holstein cows. Serum concentrations of IGF-I and ß-hydroxybutyrate (BHBA) were quantified in samples collected in sequential blood collections. Genotyping of the IGF-I and STAT5A genes was performed. The IGF-I polymorphism distribution was 35.9% CC, 46.1% CT and 18% TT. The IGF-I concentrations in circulation were greater in cows of the TT compared with both the CT and CC groups (Pâ¯<â¯0.05). Genotype had a linear association (Pâ¯<â¯0.05) with DTO and CCI, which were less for cows of the TT group. There was no association of STAT5A BstEII on serum IGF-I or reproductive variables (P> 0.05). When combining the GHR AluI T allele, obtained in a previous study, and the IGF-I SnaBI T allele from the current study, for the same cows, there were additive associations of both with serum IGF-I, BHBA, number of services per conception, DTO and CCI (Pâ¯<â¯0.05). Thus, the IGF-1 SnaBI TT appears to be associated with fewer DTO and lesser CCI of lactating dairy cows and had an additive association with the GHR AluI T allele on indicators for improvement of fertility.
Subject(s)
Cattle/physiology , Insulin-Like Growth Factor I/genetics , Insulin-Like Growth Factor I/metabolism , Polymorphism, Single Nucleotide , Receptors, Somatotropin/genetics , STAT5 Transcription Factor/genetics , Animals , Cattle/genetics , FemaleABSTRACT
Paraoxonase 1 (PON1) is a negative acute phase plasma protein synthesized by the liver that has anti-oxidant activity. The aim of this study was to evaluate the association of single nucleotide polymorphisms (SNPs) in the PON1 promoter region with plasma PON1 activity and fertility in Holstein dairy cows. Sixty-eighty Holstein cows were used in this initial investigative study. Blood samples were collected weekly beginning 28 days prior to expected calving, twice weekly in week 1 and 2 postpartum, and then once weekly through 6 weeks postpartum for plasma PON1 activity analysis. Cows were synchronized for ovulation and timed AI at 63-70 DIM using an Ovsynch program. Pregnancy diagnosis was confirmed by rectal palpation and reproductive performance data was recorded until 210 DIM. DNA was extracted from blood of each cow and a fragment of proximal PON1 gene promoter was sequenced. Seven single nucleotide polymorphisms (SNPs) were identified in the promoter region of the PON1 gene at positions -22, -105, -176, -221, -392, -611 and -676, six of which were significantly associated with plasma PON1 activity level. The SNPs -221 and -392 were significantly associated with both plasma PON1 activity and the calving to conception interval (Pâ¯<â¯0.05) with no significant effect on calving to first ovulation interval. In conclusion, the genotypes associated with higher plasma PON1 activity in SNP locations -221 and -392 were also associated with a reduced calving to conception interval in this study set of cows. These SNPs may provide novel genetic markers for improved fertility in future larger studies in dairy cows.
Subject(s)
Aryldialkylphosphatase/metabolism , Cattle/genetics , Cattle/physiology , Fertility/genetics , Polymorphism, Single Nucleotide , Animals , Antioxidants/metabolism , Aryldialkylphosphatase/genetics , Estrus Synchronization , Female , Gene Expression Regulation, Enzymologic , Genotype , Insemination, Artificial , Postpartum Period , PregnancyABSTRACT
In transition dairy cows, plasma levels of the insulin-sensitizing hormone adiponectin fall to a nadir at parturition and recover in early lactation. The transition period is also characterized by rapid changes in metabolic and hormonal factors implicated in other species as positive regulators of adiponectin production (i.e., negative energy balance, lipid mobilization) and others as negative regulators (i.e., reduced leptin and insulin and increased growth hormone and plasma fatty acids). To assess the role of onset of negative energy balance and lipid mobilization after parturition, dairy cows were either milked thrice daily (lactating) or never milked (nonlactating) for up to 4 wk after parturition. Plasma adiponectin was 21% higher across time in nonlactating than lactating cows. Moreover, nonlactating cows recovered plasma adiponectin at similar rates as lactating cows even though they failed to lose body condition. Next, we assessed the ability of individual hormones to alter plasma adiponectin in transition dairy cows. In the first experiment, dairy cows received a constant 96-h intravenous infusion of either saline or recombinant human leptin starting on d 8 of lactation. In the second experiment, dairy cows were studied in late pregnancy (LP, starting on prepartum d -31) and again in early lactation (EL, starting on d 7 postpartum) during a 66-h period of basal sampling followed by 48 h of hyperinsulinemic-euglycemia. In the third experiment, cows were studied either in LP (starting on d -40 prepartum) or EL (starting on d 7 postpartum) during a 3-h period of basal sampling followed by 5 d of bovine somatotropin treatment. Plasma adiponectin was reduced by an average of 21% in EL relative to LP in these experiments, but neither leptin, insulin, or growth hormone treatment affected adiponectin in LP or EL. Finally, the possibility that plasma fatty acids repress plasma adiponectin was evaluated by intravenous infusion of a lipid emulsion in nonpregnant, nonlactating cows in the absence or presence of glucagon for 16 consecutive hours. The intralipid infusion increased plasma fatty acid concentration from 102 to over 570 µM within 3 h but had no effect on plasma adiponectin irrespective of presence or absence of glucagon. Overall, these data suggest that energy balance around parturition may regulate plasma adiponectin but do not support roles for lipid mobilization or sustained changes in the plasma concentration of leptin, insulin, growth hormone, or fatty acids.
Subject(s)
Adiponectin/blood , Cattle/physiology , Energy Metabolism/physiology , Lactation/physiology , Adiponectin/metabolism , Animals , Female , Glucagon , Humans , Insulin/blood , Leptin/blood , Milk/metabolism , Parturition/blood , Postpartum Period/blood , PregnancyABSTRACT
The objective of this study was to determine the effects of uterine and systemic inflammatory responses to uterine bacterial contamination at calving in dairy cows on the growth and ovulatory outcomes of the first dominant follicle postpartum. Ovulatory capability of the first dominant follicle postpartum was predicted in 53 multiparous cows by using a combination of follicle growth characteristics and circulating estradiol concentrations. Endotoxin levels were assayed in follicular fluid samples that were aspirated the day after ovulatory outcome prediction. Plasma levels of haptoglobin, a proinflammatory acute phase protein, and paraoxonase, a negative acute phase protein were determined. Uterine bacteria and inflammation were evaluated in three uterine fluid samples from each cow collected on the day of calving, the day after follicle aspiration, and at 35 days postpartum. Cows that had a strong initial uterine inflammatory response (robust recruitment of polymorphonuclear leukocytes of ≥ 35% and cows with uterine pH < 8.5 on the day of calving) were more likely to have an ovulatory first dominant follicle. Follicular fluid endotoxin levels were higher in non-ovulatory cows compared with ovulatory cows. Endotoxin levels in circulation were not different between ovulatory groups but were higher prepartum than on day 7 and 14 postpartum. Systemic inflammation characterized by elevated haptoglobin concentrations was higher in non-ovulatory cows despite similar bacterial contamination and circulating endotoxin levels. Paraoxonase activity in follicular fluid was significantly associated with the paraoxonase activity in plasma, however, plasma paraoxonase concentrations were not different between non-ovulatory and ovulatory cows. Cows with a higher uterine bacterial load on the day of calving had slower ovarian follicle growth. In summary, a robust uterine inflammatory response on the day of calving was positively associated with ovarian function while elevated systemic inflammation during the early postpartum period was negatively associated with the ovulatory status of the first dominant follicle postpartum.
Subject(s)
Dairying , Ovarian Follicle/physiopathology , Postpartum Period/physiology , Uterine Diseases/veterinary , Animals , Cattle , Female , Inflammation/physiopathology , Inflammation/veterinary , Ovulation , Uterine Diseases/physiopathologyABSTRACT
The objective of the study was to evaluate the effect of prepartum and postpartum (PP) supplementation with 2 isomers of conjugated linoleic acid (CLA) on reproductive parameters and some related metabolic factors in dairy cows. High-producing, multiparous Holstein Friesian cows (n = 60) were allotted to 3 treatment groups: the CLA1 group (n = 20) was supplemented with 70 g of lipid-encapsulated CLA providing 7 g each of cis-9,trans-11 and trans-10,cis-12 CLA from d 21 (d 21) before expected calving until d 7 after artificial insemination (AI), that is, until 77 to 91 d PP; the CLA2 group (n = 20) was supplemented with the same amount of CLA beginning at calving until d 7 after AI; and the control group (n = 20) received an isocaloric, isonitrogenous, and isolipidic diet. Blood samples were taken weekly to measure glucose, insulin, insulin-like growth factor-I (IGF-I), and leptin. Liver biopsy was performed in 10 cows per group for growth hormone receptor 1A and IGF-I mRNA analyses. At d 49 to 63 PP, ovulation was synchronized with the Pre-Synch protocol followed by fixed-time AI. Milk progesterone was monitored from calving until d 35 post-AI. Cows returning to estrus following AI were inseminated. Supplementation with CLA before calving improved the recovery of plasma leptin levels in the early PP period (from the day of calving until wk 3 PP; treatment effect). Later PP (wk 5), plasma IGF-I, and leptin remained significantly higher in both CLA1 and CLA2 groups compared with control, although hepatocellular IGF-I mRNA was not different among groups. Plasma IGF-I levels remained higher in both CLA-treated groups on the day of AI. Growth hormone receptor 1A mRNA levels in hepatic tissue decreased in all groups, reaching a nadir in the first week PP. Days to first PP ovulation did not differ between groups; however, both supplemented groups conceived earlier compared with control (d 97 ± 19, d 97 ± 23, and d 113 ± 30 for CLA1, CLA2, and control, respectively). Plasma progesterone concentration was higher in both supplemented groups on d 2 to 5 following the synchronized ovulation than in controls. We concluded that CLA supplementation around calving alters PP metabolic signals as reflected by higher plasma leptin and IGF-I levels. Conjugated linoleic acid stimulated early luteal function and reduced the PP interval to conception.
Subject(s)
Cattle , Growth Hormone/drug effects , Insulin-Like Growth Factor I/drug effects , Linoleic Acids, Conjugated/administration & dosage , Reproduction/drug effects , Animals , Female , Lactation , Lipids , Milk , Postpartum Period , PregnancyABSTRACT
Paraoxonase-1 (PON1) is an enzyme found in serum and follicular fluid that protects cell membrane and circulating lipids against oxidative damage. The aims of this study were to measure the direct effects of recombinant PON1 (rPON1) on bovine oocyte maturation at the molecular level (gene expression) and to measure the carry-over effects of PON1 on pre-implantation embryo development in vitro. COCs were submitted to IVM with the addition of 0.0, 0.02, 0.04 and 0.08 mg ml(-1) of rPON1, corresponding to an average PON1 arylesterase enzyme activity of 2.2 ± 0.4, 15.5 ± 1.5, 30.2 ± 3.0 and 57.9 ± 5.0 U ml(-1) , respectively. The results indicated that addition of rPON1 during IVM improved embryo development in a dose-dependent manner as D7 embryo development was 22.2%, 29.4%, 32.2% and 37.0% for the treatment groups, respectively (p = 0.02). In conclusion, addition of PON1 enzyme during IVM exerted dose-related positive effects on embryo development rates to blastocysts.
Subject(s)
Aryldialkylphosphatase/pharmacology , Cattle/embryology , Embryo Culture Techniques/veterinary , In Vitro Oocyte Maturation Techniques/veterinary , Animals , Culture Media , Cumulus Cells/drug effects , Embryo, Mammalian/drug effects , Embryonic Development/drug effects , Gene Expression Regulation, Developmental/drug effectsABSTRACT
Conjugated linoleic acid (CLA) isomers can affect the lipid profile and signaling of cells and thereby alter their function. A total of 5,700 bovine oocytes were used in a structured series of experiments to test the effects of CLA cis-9,trans-11 and CLA trans-10,cis-12 in vitro. In experiment 1, high doses of each CLA isomer during in vitro maturation (IVM) were compared with high or low doses during the entire in vitro culture (IVC) of parthenogenetic embryos. High doses of the CLA isomers ranged from 50 to 200 µM and low doses were 15 and 25 µM. In experiment 2, the low doses of each CLA isomer were tested during IVM/IVC on embryos produced by in vitro fertilization (IVF). Experiment 3 compared the effects of 15 µM doses of each CLA isomer during IVM or IVC of IVF embryos. In experiment 4, post-rewarming survival rates and blastomere counts were assessed for embryos supplemented with each CLA isomer during IVM or for 36 h before vitrification. In experiment 1, when either CLA isomer was provided only during IVM, we observed no effects on overall rates of maturation, cleavage, or blastocysts (92.2 ± 1.6%, 78.3 ± 4.1%, and 28.9 ± 5.1%, respectively). However, high doses of each CLA isomer, but not low doses, during the entire embryo culture period decreased blastocyst rates (5-20%) in a dose-dependent manner. Cleavage rates improved with 15 or 50 µM CLA trans-10,cis-12. Progesterone concentrations in maturation media were significantly increased by high doses of each CLA isomer compared with control, but low doses of CLA isomers had no effect. In experiment 2 with IVF embryos, low doses of each CLA isomer did not alter cleavage rates (average 84.9 ± 1.9%) and only 25 µM CLA trans-10,cis-12 during IVC reduced blastocyst rates below those of controls (25.5 ± 2.1 vs. 38.2 ± 2.3%). The lipid content of embryos was increased and relative expression of the BIRC5 (baculoviral IAP repeat containing 5) gene was depressed by CLA trans-10,cis-12. In experiment 3, low doses (15µM) of each CLA isomer during IVC significantly reduced blastocyst rates (20.6 ± 2.4% and 27.7 ± 1.2% vs. 34.18 ± 1.2% for CLA trans-10,cis-12 and CLA cis-9,trans-11 compared with control, respectively) with less effect of each CLA during IVM. In experiment 4, adding 100 µM CLA cis-9,trans-11 during the final 36 h of culture resulted in a high survival rate after rewarming and culture, and the higher embryo blastomere count was comparable to that of control embryos not undergoing vitrification. In conclusion, supplementation with either CLA isomer did not improve embryo production, but inclusion of CLA cis-9,trans-11 before vitrification improved the quality of bovine IVF embryos after rewarming and culture.
Subject(s)
Cattle/embryology , Cryopreservation/veterinary , Fertilization in Vitro/veterinary , Linoleic Acids, Conjugated/pharmacology , Animals , Blastocyst/physiology , Cryopreservation/methods , Embryo Culture Techniques/methods , Embryo Culture Techniques/veterinary , Embryonic Development/drug effects , In Vitro Oocyte Maturation Techniques/methods , In Vitro Oocyte Maturation Techniques/veterinary , Isomerism , Linoleic Acids, Conjugated/chemistry , Lipids/analysis , Oocytes/physiology , Progesterone/analysis , Progesterone/physiologyABSTRACT
The objective was to evaluate the individual and combined effect of anovulation and cytological endometritis (CTE) on the reproductive performance of dairy cows. A total of 1,569 cows from 3 data sets were used. In data set 1, 403 Holstein cows from 5 dairies in New York were used. In data set 2, 750 Holstein cows from 2 dairies, one in Florida and one in California were used. In data set 3, 416 dairy cows, 165 Holsteins, 36 Jerseys, and 215 Holstein-Jersey crossbreeds from a grazing dairy in Florida were used. Cyclicity and CTE was determined at 35±3 (data set 2) or 49±3 d in milk (data sets 1 and 3). A variable (VarCycCTE) containing all 4 possible permutations between cyclicity (cyclic = Cyc; anovular = Anov) and CTE (present = CTE; absent = Healthy) was created. In the combined data set (sets 1, 2, and 3), pregnancy per artificial insemination (P/AI) diagnosed at 30 to 38 d after first AI was affected by VarCycCTE, with AnovCTE cows having decreased P/AI compared with CycHealthy cows (21.3 vs. 46.7%), whereas AnovHealthy (37.9%) and CycCTE cows (36.0%) had intermediate P/AI. Pregnancy per artificial insemination for the individual data sets and for pregnancy diagnosed at 63 to 74 d after artificial insemination followed a similar pattern. Pregnancy loss was not affected by VarCycCTE. Hazard of pregnancy up to 300 d in milk was affected by VarCycCTE in the combined data sets 1 and 2, with AnovCTE [hazard ratio (HR)=0.55], AnovHealthy cows (HR=0.71), and CycCTE (HR=0.8) having decreased hazard of pregnancy compared with CycHealthy cows. Median days open were 200, 159, 145, and 121 for AnovCTE, AnovHealthy, CycCTE, and CycHealthy, respectively. Hazard of pregnancy for the individual data sets followed a similar pattern. In summary, both anovulation and CTE were negatively associated with reproductive performance and, when combined, they had an additive negative effect.
Subject(s)
Anovulation/veterinary , Cattle Diseases/physiopathology , Cattle , Endometritis/veterinary , Reproduction , Animals , California , Diet/veterinary , Female , Florida , Housing, Animal , Insemination, Artificial/veterinary , Milk , New York , Pregnancy , Retrospective StudiesABSTRACT
Despite advantages of in vitro embryo production in many species, widespread use of this technology is limited by generally lower developmental competence of in vitro derived embryos compared to in vivo counterparts. Regardless, in vivo or in vitro gametes and embryos face and must adjust to multiple microenvironments especially at preimplantation stages. Moreover, the embryo has to be able to further adapt to environmental cues in utero to result in the birth of live and healthy offspring. Enormous strides have been made in understanding and meeting stage-specific requirements of preimplantation embryos, but interpretation of the data is made difficult due to the complexity of the wide array of culture systems and the remarkable plasticity of developing embryos that seem able to develop under a variety of conditions. Nevertheless, a primary objective remains meeting, as closely as possible, the preimplantation embryo requirements as provided in vivo. In general, oocytes and embryos develop more satisfactorily when cultured in groups. However, optimization of individual culture of oocytes and embryos is an important goal and area of intensive current research for both animal and human clinical application. Successful culture of individual embryos is of primary importance in order to avoid ovarian superstimulation and the associated physiological and psychological disadvantages for patients. This review emphasizes stage specific shifts in embryo metabolism and requirements and research to optimize in vitro embryo culture conditions and supplementation, with a view to optimizing embryo culture in general, and culture of single embryos in particular.
Subject(s)
Animals, Domestic/embryology , Blastocyst/metabolism , Embryo Culture Techniques , Animals , Antioxidants/metabolism , Blastocyst/physiology , Culture Media , Embryonic Development , Female , Fertilization in Vitro , Humans , Oocytes/cytology , Oxidative StressABSTRACT
The aim of this work was to determine the effects of a growth hormone receptor (GHR) AluI polymorphism on the reproductive performance of Holstein cows. The cows (n = 94) were on the study from 3 weeks prepartum until 210 days in milk (DIM). Blood samples were collected at -21, 0, 7, 21, and 60 DIM. For GHR genotyping, DNA was extracted from blood and the presence of the alleles determined after polymerase chain reaction and digestion with the restriction enzyme AluI. Milk samples were collected for progesterone analysis and detection of ovulation until first breeding. Cows were submitted to an OvSynch-TAI protocol at 55 DIM that was repeated for cows diagnosed as not pregnant. Data were analyzed with SAS for polynomial effects of the presence of 0, 1, or 2 GHR AluI (-) alleles. Among the cows, 37% had the AluI(+/+) genotype, 51% had AluI(-/+), and 12% were AluI(-/-). Interval from calving to first ovulation was not different among genotypes (P > 0.05). Cows carrying at least one GHR AluI(-) allele had fewer number of services per conception (P = 0.02). In addition, there was a linear reduction (P = 0.02) in the calving to conception interval among genotypes with fewest days for GHR AluI(-/-) cows. GHR AluI(-/-) cows also had the highest serum IGF-I concentrations (P = 0.03). Milk production and composition were not different among genotypes (P > 0.05). The presence of one or two GHR AluI(-) alleles in Holstein cows was associated with a linear reduction in the calving to conception interval and a reduction in the number of AI/conception.
Subject(s)
Cattle/physiology , Fertility/genetics , Polymorphism, Genetic , Animals , Genotype , Insulin-Like Growth Factor I/chemistry , Insulin-Like Growth Factor I/genetics , Insulin-Like Growth Factor I/metabolism , Milk/chemistry , Progesterone/metabolism , Receptors, Somatotropin/geneticsABSTRACT
Normal metabolic activity in ovarian follicles may result in oxidative stress and damage to oocytes. The aim of this study was to evaluate expression of the natural anti-oxidants paraoxonase (PON) 1, 2 and 3 in granulosa cells and PON1 activity in follicular fluid (FF) and plasma of dairy cows. For the first experiment, ovaries were collected from cows at slaughter, after which follicles were dissected and classified as oestrogen active (EAF) or atretic (ATF). Expression of PON1, PON2 and PON3 mRNA was evaluated in granulosa cells, and activity of PON1 was measured in FF. PON1 mRNA was undetectable in granulosa cells, PON2 mRNA expression was not different between follicle types, and PON3 mRNA tended to be higher in EAF (p = 0.11). The activity of PON1 in FF was higher (p = 0.01) for EAF (82.6 ± 8.0 kU/L) than ATF (53.9 ± 6.8 kU/L), as were high-density lipoproteins (HDL), low-density lipoproteins (LDL) and total cholesterol concentrations. In the second experiment, we aimed to compare plasma and FF PON1 activity in early lactation Holstein cows (n = 15) with pre-ovulatory EAF. Activity of PON1 was twofold higher (p < 0.0001) in plasma (122.5 ± 11.1 kU/L) than in FF (61.4 ± 5.2 kU/L). Plasma concentrations were also higher (p < 0.0001) for HDL, LDL and total cholesterol when compared to FF. In conclusion, FF concentrations of PON1, HDL, LDL and total cholesterol were higher in healthy oestrogen active bovine follicles than in atretic follicles. PON1 was not expressed by granulosa cells indicating that high PON1 activity in bovine FF is apparently derived by transfer from blood in association with HDL.
Subject(s)
Aryldialkylphosphatase/metabolism , Cattle/metabolism , Follicular Fluid/enzymology , Gene Expression Regulation, Enzymologic/physiology , Granulosa Cells/enzymology , Animals , Aryldialkylphosphatase/blood , Aryldialkylphosphatase/genetics , Cholesterol/blood , Cholesterol/metabolism , Dairying , Female , Granulosa Cells/metabolismABSTRACT
The aim of this work was to monitor the pattern of serum acute phase proteins [paraoxonase (PON), haptoglobin (Hp) and albumin] during the peripartum period of normal healthy cows (n=16) compared to that in cows diagnosed with uterine infection (n=15). Albumin concentrations were lower at 21 days before expected calving in cows subsequently diagnosed with uterine infection and predicted the occurrence of uterine infection with an accuracy of 79.3%. Cows diagnosed with uterine infection had a postpartum reduction in serum PON activity and the concentration of Hp increased at 7 DIM for multiparous uterine infected cows. In conclusion, cows diagnosed with uterine infection had reduced serum albumin concentration 21 days before calving, lower PON activity at 7 days after calving, and increased Hp in multiparous cows at 7 DIM compared to healthy cows.
Subject(s)
Aryldialkylphosphatase/blood , Cattle Diseases/blood , Uterine Diseases/veterinary , Animals , Cattle , Female , Haptoglobins/metabolism , Postpartum Period , ROC Curve , Serum Albumin, Bovine/metabolism , Uterine Diseases/bloodABSTRACT
The objective of this study was to evaluate the association between uterine disease and indicators of neutrophil (PMN) and systemic energy status in dairy cows. Peripheral blood (120 mL) was collected weekly from 84 Holstein cows for PMN isolation and plasma collection from calving until 42 d in milk (DIM). The final analysis included 80 cows. Of those, 20 cows were classified as having metritis (fetid uterine discharge and fever), 15 as having subclinical endometritis (SCE; >or=10% PMN on uterine cytology), and 45 as healthy controls. Plasma haptoglobin concentration was increased only in cows that developed metritis. Neutrophil glycogen content was reduced in cows developing metritis compared with healthy cows on the day of calving and at 7 and 42 DIM. Cows with SCE had lower PMN glycogen content than healthy cows at 7, 28, and 42 DIM. Blood glucose was affected by disease status within parity. Primiparous metritis cows had greater blood glucose concentrations than healthy primiparous cows. Multiparous metritis cows tended to have lower blood glucose concentration than multiparous SCE cows. Cows that developed metritis and SCE had or tended to have greater NEFA and BHBA than healthy cows, mainly around calving. At calving, cows that developed metritis had higher plasma estradiol concentration than healthy cows and greater plasma cortisol than cows that had SCE. Plasma insulin was not affected. Plasma glucagon was increased for SCE cows. Cows that developed uterine disease experienced a greater degree of negative energy balance and had decreased lower intracellular PMN glycogen levels, which could be a major predisposing factor for disease because of decreased availability of oxidative fuels.
Subject(s)
Cattle Diseases/physiopathology , Energy Metabolism , Neutrophils/metabolism , Uterine Diseases/veterinary , 3-Hydroxybutyric Acid/blood , Animals , Blood Glucose/analysis , Cattle , Cattle Diseases/blood , Dairying , Estradiol/blood , Fatty Acids, Nonesterified/blood , Female , Glycogen/blood , Haptoglobins/analysis , Time Factors , Uterine Diseases/blood , Uterine Diseases/physiopathologyABSTRACT
Our objectives were to determine the effects of early ovulation on fertility and uterine health of dairy cows. Four hundred and forty-five Holstein cows (185 primiparous and 260 multiparous) from five herds were used. Blood samples were collected at 21, 35 and 49 days in milk (DIM) and cows were considered to be cyclic at 21 DIM (Cyc21) if serum progesterone (P4) concentration was above 1 ng/ml, cyclic by 49 DIM (Cyc49) if P4 concentration was above 1 ng/ml at 35 or 49 DIM, or not cyclic (NotCyc) if P4 concentration was below 1 ng/ml at all sample times. Endometrial cytology for diagnosis of subclinical endometritis was examined at 49 DIM in a subset of 414 cows. Cows in the group Cyc21 had increased hazard of insemination, for the first service, compared with cows in Cyc49 [hazard ratio (HR) = 1.40; 95% CI = 1.10-1.79; p = 0.006] and NotCyc (HR = 2.07; 95% CI = 1.52-2.82; p < 0.001). Cows in the Cyc49 group also had increased hazard of insemination compared with cows in the NotCyc group (HR = 1.47; 95% CI = 1.13-1.93; p = 0.005). Median days to insemination were, respectively, 71, 76 and 96 for cows in Cyc21, Cyc49 and NotCyc groups. Cows in Cyc21 had greater first service pregnancy per AI than Cyc49 [38.6 vs 28.1%; adjusted odds ratio (AOR) = 1.67; 95% CI = 1.01-2.75; p = 0.04] and NotCyc (38.6 vs 23.6%; AOR = 2.08; 95% CI = 1.08-4.00; p = 0.03). Pregnancy per AI was similar in Cyc49 and NotCyc cows (28.1 vs 23.6%; AOR = 1.25; 95% CI = 0.70-2.24; p = 0.45). Cows in Cyc21 had increased hazard of pregnancy up to 300 DIM compared with Cyc49 (HR = 1.52; 95% CI = 1.17-1.96; p = 0.002) and NotCyc (HR = 1.98; 95% CI = 1.41-2.78; p < 0.001). Cows in Cyc49 tended to have increased hazard of pregnancy compared with NotCyc (HR = 1.31; 95% CI = 0.96-1.77; p = 0.09). Median days to pregnancy were, respectively, 103, 147 and 173 for cows in Cyc21, Cyc49 and NotCyc groups. Cows in the Cyc21 group had decreased prevalence of subclinical endometritis compared with cows in the NotCyc group (29.9 vs 43.7%; AOR = 0.53; 95% CI = 0.29-0.97; p = 0.04); however, the prevalence did not differ from the Cyc49 group (29.9 vs 39.1%; AOR = 0.68; 95% CI = 0.41-1.14; p = 0.15). Cyc49 cows had similar prevalence of subclinical endometritis compared with NotCyc cows (AOR = 0.77; 95% CI = 0.46-1.29; p = 0.32). Early postpartum ovulation was associated with improved uterine health and fertility.
Subject(s)
Cattle/physiology , Dairying , Fertility/physiology , Ovulation/physiology , Postpartum Period/physiology , Animals , Dinoprost/administration & dosage , Dinoprost/pharmacology , Drug Administration Schedule , Female , Fertility Agents/administration & dosage , Fertility Agents/pharmacology , Lactation , Ovulation/drug effects , Progesterone/blood , Time FactorsABSTRACT
The objectives were to determine the effects of PGF(2alpha) treatment on the prevalence of subclinical endometritis (SCE) and fertility of dairy cows. A total of 406 Holstein cows (167 primiparous and 239 multiparous) from 5 herds were used. Uterine lavage for diagnosis of SCE, PGF(2alpha) treatment, evaluation of body condition scores (BCS), and collection of blood samples for estrous cyclicity determination were performed at 21, 35, and 49 d in milk (DIM). Polymorphonuclear cells (PMN) were quantified and thresholds for diagnosing SCE were selected by receiver operating characteristics analysis. Cows classified as having SCE at 35 DIM (>or=6.5% PMN) and 49 DIM (>or=4.0% PMN) had increased time to pregnancy; however, cows classified as having SCE at 21 DIM (>or=8.5% PMN) did not. Median days to pregnancy were delayed by 30 (151 vs. 121 d) and 40 (169 vs. 129) d for cows classified as having SCE at 35 and 49 DIM, respectively. Treatment with PGF(2alpha) did not affect the prevalence of SCE either at 35 (37.9 vs. 38.4%) or at 49 DIM (34.0 vs. 40.4%). Treatment with PGF(2alpha) did not affect time to first insemination (AI; median 76 DIM for cows treated with PGF(2alpha); 79 DIM for control. Nonetheless, PGF(2alpha) treatment increased pregnancy to first AI in all the cows (35.5 vs. 24.1%) and hazard ratio (HR) of pregnancy in cows with BCS Subject(s)
Cattle Diseases
, Dinoprost/administration & dosage
, Endometritis/complications
, Fertility/drug effects
, Infertility, Female/veterinary
, Animals
, Cattle
, Cattle Diseases/drug therapy
, Cattle Diseases/prevention & control
, Corpus Luteum/physiopathology
, Endometritis/diagnosis
, Endometritis/prevention & control
, Female
, Infertility, Female/drug therapy
, Insemination, Artificial/veterinary
, Lactation
, Pregnancy
, Progesterone/blood
ABSTRACT
Thiazolidinediones (TZD) are potent synthetic ligands for peroxisome proliferator-activated receptor-gamma that have been shown previously to reduce plasma nonesterified fatty acids and increase peripartal dry matter intake (DMI) in dairy cows. Data from Holstein cows (n = 36) entering their second or greater lactation were used to determine whether late prepartum administration of TZD would affect periparturient metabolism, milk production, and ovarian activity. Cows were administered 0, 2.0, or 4.0 mg of TZD/kg of BW by intrajugular infusion once daily from 21 d before expected parturition until parturition. Plasma samples were collected daily from 22 d before expected parturition through 21 d postpartum and twice weekly from wk 4 through 9 postpartum. In response to increasing TZD dosage, plasma nonesterified fatty acid concentrations decreased linearly during the postpartum period (d 0 to +21: 348, 331, 268 +/- 31 microEq/L, respectively). Plasma concentrations of glucose were highest in cows administered 4.0 mg of TZD/kg of BW during the peripartum and postpartum periods (d -7 to +7: 57.9, 57.8, 61.1 +/- 0.8 mg/dL and d 0 to +21: 51.6, 49.3, 54.7 +/- 1.1 mg/dL, respectively). Plasma concentrations of beta-hydroxybutyrate were increased during the peripartum period by TZD administration (9.6, 9.9, 10.2 +/- 0.3 mg/dL) but were not affected during the postpartum period. Plasma insulin was not affected by treatment during any time period. Postpartum liver triglyceride content was decreased linearly (11.0, 10.4, 4.2 +/- 1.6%) and glycogen content was increased linearly (2.16, 2.38, 2.79 +/- 0.19%) by prepartum TZD administration. Prepartum TZD administration linearly increased DMI during the peripartum period (d -7 to +7: 16.1, 17.2, 17.3 +/- 0.5 kg/d). Cows administered TZD prepartum maintained higher postpartum body condition scores than control cows (wk 1 through 9: 2.77, 2.89, 3.02 +/- 0.05). There was no effect of prepartum TZD on milk yield; however, yields of 3.5% fat-corrected milk (52.2, 54.6, 48.0 +/- 1.6 kg/d) and most other milk components were decreased in cows that received 4.0 mg of TZD/kg of BW prepartum. Prepartum TZD administration linearly decreased the number of days to first ovulation (29.3, 28.3, 19.0 +/- 3.6 d). These results suggest that prepartum administration of TZD improves metabolic health and DMI of periparturient dairy cows and may decrease reliance on body fat reserves during early lactation.
Subject(s)
Cattle/metabolism , Lactation/drug effects , Ovary/drug effects , Thiazolidinediones/pharmacology , Animals , Dairying , Fatty Acids, Nonesterified/blood , Female , Insulin/blood , Liver/chemistry , Liver/drug effects , Pregnancy , Reproduction/drug effects , Time FactorsABSTRACT
The feeding of conjugated linoleic acid (CLA) supplements to early-lactation dairy cows has been shown to decrease milk fat synthesis and possibly improve reproductive performance. However, previously reported studies used too few animals to clearly establish the effect of CLA on reproduction. Our objective was to combine data from these studies to evaluate the association of CLA with time to first ovulation and time to conception using methods of survival analysis and overall success of pregnancy by logistic regression. A database was compiled of individual animal data (n = 212) from 5 controlled studies in which CLA had been supplemented to early-lactation dairy cows. Survival analysis incorporated both semi-parametric models (Cox proportional hazards) and parametric models (log-normal). The probability of cows becoming pregnant increased in a nonlinear manner as trans-10, cis-12 CLA dose increased, with the optimal dose predicted to be 10.1 g/d. At the optimal dose, the probability of pregnancy was increased by 26% compared with those animals receiving no CLA (probability = 91% and 72%, respectively). Similarly, the log-normal model predicted that time to conception was decreased in a nonlinear manner with increasing trans-10, cis-12 CLA dose. The predicted optimal dose was 10.5 g of trans-10, cis-12 CLA/d and at this dose the median time to conception was decreased by 34 d when compared with those cows not receiving CLA (117 vs. 151 d in milk, respectively). The log-normal model was also the best-fit model for time to first ovulation. Overall, this multi-study analysis demonstrated a strong concordance between the nature of the dose response and the predicted optimal dose of trans-10, cis-12 CLA across the 3 reproductive variables evaluated. These results indicate that reproductive performance of dairy cows may be improved by feeding of CLA supplements during early lactation.
Subject(s)
Cattle/physiology , Dietary Supplements , Lactation/physiology , Linoleic Acids, Conjugated/administration & dosage , Reproduction/physiology , Animals , Female , Logistic Models , Pregnancy , Survival AnalysisABSTRACT
Multiparous Holstein cows (n=58) were used to study the effects of peripartum dietary supplementation on metabolic status, liver function and reproduction variables. Diets for cows were as follows: (a) no supplementation (CTL), (b) prilled fatty acids as 1.9% of DM (PrFA), (c) calcium salts of long chain n-6 fatty acids as 2.24% of DM (CaLFA) or (d) daily topdressing with 769 g of 65% propylene glycol (PGLY). Supplements were fed during the last 21 days before expected calving except for PGLY that continued until 21 days after parturition. Ovarian activity was monitored by transrectal ultrasonography and days to first ovulation were recorded. Liver biopsies were obtained on day 8 and 21 postpartum and analyzed for triglyceride content and mRNA expression of pyruvate carboxylase, cytosolic phosphoenolpyruvate carboxykinase, carnitine palmytoyltransferase 1A, and peroxisome proliferator-activated receptor-alpha. At 71 days following parturition, stage of ovarian cycles was synchronized and at day15 of the cycle oxytocin was injected i.v., blood samples were obtained at frequent intervals, and analyzed for 13,14 dihydro, 15-keto PGF(2alpha) (PGFM). Milk production and milk components were not different among treatment groups. Cows in PGLY gained body condition score (BCS) prepartum and net energy balance prepartum tended to be greater, but was not different postpartum from other groups. PGLY supplementation increased plasma insulin concentration prepartum, but not during the postpartum period. No significant differences were observed in plasma concentrations of glucose, NEFA, and insulin-like growth factor or hepatic triglyceride content, but all supplements tended to decrease beta hydroxybutyrate postpartum compared to CTL cows. Abundance of mRNA of gluconeogenic and lipid oxidation genes was not different among treatment groups. Days to first ovulation and uterine PGF(2alpha) production in response to an oxytocin treatment were not significantly different among treatment groups. Peripartum supplementation did not result in the substantial improvement of metabolic profile in early lactation nor significantly affect days to first ovulation and PGFM response to an oxytocin treatment.