Messenger RNA expression of angiotensin-converting enzyme, endothelin, cyclooxygenase-2 and prostaglandin synthases in bovine placentomes during gestation and the postpartum period

The bovine placenta contains local vasoactive-related systems, including angiotensin-converting enzyme (ACE), endothelin-1 (ET-1), ET-A receptor (ETAR) and ET-B receptor (ETBR), as well as arachidonic acid (AA) cascade-related enzymes, such as cyclooxygenase-2 (Cox-2), prostaglandin E-synthase (PGES) and prostaglandin F-synthase (PGFS). The mRNA expression of these molecules was examined in bovine placentomes (caruncles and cotyledons) collected immediately (0 h) and 6h after spontaneous parturition from 15 cows with early (fetal membranes released within 6 h of parturition) or late (fetal membranes released 6-12 h after parturition) detachment, as well as from 15 pregnant cows at a slaughterhouse. Significant differences were observed in expression of ET-1, ETAR and ETBR mRNAs between gestation and the postpartum period in both caruncles and cotyledons. Significant differences were also found between 0 and 6 h postpartum in the expression of ETBR mRNA in the early detachment group and PGES mRNA in the early and late detachment groups. Compared to PGFS, both Cox-2 and PGES exhibited opposite mRNA expression patterns during gestation and the postpartum period. The vasoactive-related peptide systems and AA cascade-related enzymes may mediate placental development and fetal membrane detachment after parturition in cattle.     

Prepartum maternal plasma glucose concentrations and placental glucose transporter mRNA expression in cows carrying somatic cell clone fetuses

In this study, the plasma glucose concentrations of cows carrying a somatic cell clone fetus during late pregnancy and placental glucose transporter (GLUT) mRNA levels at parturition were examined. Parturition was induced using dexamethasone, prostaglandin F(2α) and estriol in cows bearing a clone (Clone) or a fetus fertilized in vivo as a control (DEX). Plasma glucose concentrations were measured in the cows (days 257 and 271 of pregnancy and at parturition) and newborn calves. Cotyledon and caruncle tissues removed just after parturition were used for mRNA extraction. Expression of mRNA was also analyzed in control cows that were induced to undergo parturition without dexamethasone (PG) or that spontaneously delivered (SP). The glucose concentrations of the Clone group were significantly low at all points examined, but those of the calves were normal. The increase in the maternal glucose concentration from day 257 to parturition was significantly lower in the Clone group. Glucose concentrations were negatively correlated with birth weight for clones (day 257; r=-0.584, day 271; r=-0.286, parturition; r=-0.549). There was no difference in mRNA levels in the cotyledons among the animals examined. In the caruncles, the Clone and PG groups showed significantly higher GLUT1 and GLUT3 mRNA levels than the SP group, and the GLUT3 mRNA level was significantly higher in the Clone group than in the DEX group. The glucocorticoid receptor α mRNA level was significantly lower in the SP group than in the DEX group. Although spontaneous parturition and administration of dexamethasone suppressed the placental GLUT mRNA levels, the action was not observed in clone pregnancy. These results raise the possibility of facilitation of glucose transportation through the placenta to meet increased nutritional requirements of overgrown clone fetuses.     

天花粉蛋白对黄牛产后胎衣脱落及受胎率的影响

以淮北黄牛为对象，在正常分娩后不同时间，用生理盐水溶解天花粉蛋白胶囊，而后灌注入黄牛子宫粘膜与胎衣之间，观察其对胎衣脱落及受胎率的影响。结果，分娩后４，１２ｈ给药组及对照组，用药后胎衣脱落时间分别为４．５９，９．１５ｈ及５６．０８ｈ；胎衣完整率为９０．７％，８７．８％及６８．６％；产后首次发情时间平均为６０．７５，６６．２９ｄ及７４．７５ｄ；试验组与对照组差异极显著（Ｐ＜０．０１）。两试验组与对照组比较，１次受胎率分别提高３３．９８％和２７．６７％；２次受胎率提高３７．９４％和２８．５８％；３次受胎率提高２３３．２９％和１９９．９４％。结果显示，天花粉蛋白能促进胎衣及时完整地排出，并显著提高受胎率     

Peripartal changes in serum alkaline phosphatase activity and lactate dehydrogenase activity in dairy cows.

Peripartal serum alkaline phosphatase activity and lactate dehydrogenase activity were measured in 30 dairy cows in order to examine the association between retained fetal membranes and enzyme activity. Daily blood samples were obtained from pregnant cows, starting 15 days before the expected day of calving until eight days after parturition. Sera from 15 cows which retained fetal membranes longer than 24 hours and 15 cows which shed fetal membranes within six hours after parturition were analyzed for alkaline phosphatase and lactate dehydrogenase enzyme activities. Mean alkaline phosphatase enzyme activities ranged from 15.93 to 32.6 U/L in retained and nonretained placenta cows. There was a trend towards higher serum alkaline phosphatase activities in retained placenta cows but the differences were not significant among the groups (P greater than 0.05). Mean lactate dehydrogenase activities ranged from 307.2 to 438.86 U/L in nonretained and retained placenta cows. Lactate dehydrogenase enzyme activities in nonretained and retained placenta cows were similar (P greater than 0.05). The alkaline phosphatase and lactate dehydrogenase enzyme activities peaked at the time of parturition in both groups. However, the differences in alkaline phosphatase and lactate dehydrogenase activities on different days within non-retained and retained placenta cows were significant (P less than 0.05). Results indicate that prepartal changes in alkaline phosphatase and lactate dehydrogenase enzyme activities are not predictive of placental retention postpartum.     

Effect of oral administration of a calcium chloride gel on blood mineral concentrations, parturient disorders, reproductive performance, and milk production of dairy cows with retained fetal membranes.

OBJECTIVE: To assess the effect of oral administration of CaCl2 gel on blood mineral concentrations, parturient disorders, reproductive performance, and milk production of dairy cows with retained fetal membranes (RFM). DESIGN: Randomized field trial. ANIMALS: 20 cows that calved normally and were not treated with CaCl2 gel (group 1), 20 cows with RFM that were treated with CaCl2 gel (group 2), and 20 cows with RFM that were not treated with CaCl2 gel (group 3). PROCEDURE: Group-2 cows were treated orally with CaCl2 gel (54 g of calcium) 24 and 48 hours after parturition. RESULTS: Administration of CaCl2 gel 24 and 48 hours after parturition did not have a significant effect on serum normalized calcium, total calcium, magnesium, or phosphorus concentrations or on incidence of metritis or left displacement of the abomasum, days to first insemination, pregnancy status after first insemination, or milk production. CONCLUSIONS AND CLINICAL RELEVANCE: Administration of CaCl2 gel 24 and 48 hours after parturition did not have significant effect on blood mineral concentrations, parturient disorders, reproductive performance, and milk production in dairy cows with RFM.     

Premature bovine parturition induced by ponderosa pine: effects of pine needles, bark and branch tips

Three components of Ponderosa pine trees were administered via gavage to pregnant cows to determine their ability to induce premature parturition. When tips of branches, bark or needles were administered (2.7 kg/day) starting at 250 d of pregnancy, early parturition was induced in an average of 3.4, 5.0 and 9.1 d, respectively, compared to 31.8 d for water gavaged controls (P less than 0.01). Less pine material was required to induce parturition with tips and bark (9.4 and 11.2 kg/cow, respectively) than with needles (25.0 kg/cow P less than 0.05). Cows fed pine material retained fetal membranes longer after parturition; 13.0 d for needles, 10.5 d for bark and greater than 3.0 d for tips versus less than 0.5 d for controls (P less than 0.01). However, 2 of the cows fed tips died within 48 hours after parturition with fetal membranes intact and the other 2 cows fed tips became depressed, pyrexic and emaciated. They survived after extended veterinary care with antibiotic and fluid therapy. Administration of pine materials modified serum progesterone concentration profiles; there was significant elevation in progesterone 1-3 days after pine material was administered. Subsequently, pine material induced a premature decline in progesterone levels prior to parturition, which occurred more rapidly in cows fed bark and tips suggesting a dose response relationship. Pine material also modified serum cortisol concentration profiles; the main difference was a prolonged elevation in cows receiving tips. Branch tips and bark from Ponderosa pine are more potent in inducing parturition than needles.     

Effect of injecting collagenase into the uterine artery during a caesarean section on the placental separation of cows induced to calve with dexamethasone

A caesarean section was performed on 30 cows before normal term and 16 to 20 hours after the induction of parturition with dexamethasone. During the surgical procedure, 20,000 U of bacterial collagenase was injected into the uterine artery of 15 of the cows. The average periods of retention of the fetal membranes were 40 hours in the treated cows and 114 hours in the control cows (P<0.001). At 36 hours after the surgery six of the treated cows (40 per cent) but all 15 of the control cows had retained fetal membranes. The collagenase-treated cows showed no abnormal clinical signs during the 10 days after the operation.     

Plasma mineral and energy metabolite concentrations in dairy cows fed an anionic prepartum diet that did or did not have retained fetal membranes after parturition

OBJECTIVE: To compare plasma total calcium, phosphorus, magnesium, nonesterified fatty acids (NEFA), beta hydroxy butyrate (BHB), and glucose concentrations in parturient dairy cows that were fed an anionic prepartum diet between those with and without retained fetal membranes (RFM) at 24 hours after parturition. ANIMALS: 152 Holstein cows that calved during October through December of 1997 PROCEDURE: All cows were fed an anionic prepartum diet. Blood sample was taken within 6 hours after parturition from randomly selected cows. Thirty-nine cows had a diagnosis of RFM at 24 hours after parturition; 113 were not affected with RFM. At calving, body condition score (BCS; 1 [thin] to 5 [obese]), parity, and calving difficulty score were recorded. Plasma calcium, phosphorus, magnesium, NEFA, BHB, and glucose concentrations were compared between cows with or without RFM. RESULTS: Cows with RFM had significantly lower plasma calcium concentration soon after calving, compared with cows without RFM. Cows with a parity of > or = 3 had significantly lower plasma concentrations of calcium and higher concentrations of magnesium, compared with cows with a parity of 1 or 2. Cows with a BCS of > or = 3.25 at calving had significantly higher plasma concentrations of BHB than cows with a BCS of 2.75 to 3.0. Cows with dystocia had significantly higher plasma concentrations of glucose, compared with cows without dystocia. CONCLUSIONS AND CLINICAL RELEVANCE: In parturient cows fed a prepartum anionic diet, those with RFM have lower plasma calcium concentrations than cows without RFM, although this association does not prove a cause-effect relationship.     

Post-partum leucocytic activity and its relationship to caesarian section and retained placenta

The activity of the leucocytes of 30 cows during 10 days after parturition is described. A comparison is made between the activity of leucocytes of 10 cows after normal parturition, 10 cows after caesarian section and 10 cows with retained placenta. When compared with normal parturition, caesarian section does not have a negative influence on the activity of the leucocytes during the puerperal period, during which time however the activity of leucocytes of cows with retained placenta is strongly decreased.     

Metabolic changes in cows with or without retained fetal membranes in transition period

The purpose of the study was to evaluate the effect of retained fetal membranes (RFM) on serum minerals and energy- and protein-related metabolites in dairy cows at a herd with a recent history of fatty liver syndrome. Forty-seven multiparous Holstein cows were selected during transition period. Nine cows had RFM longer than 24 h after calving. Blood samples were obtained on prepartum days 21 and 7 and postpartum days 7 and 21. We used repeated measure procedure of anova to evaluate the effect of RFM on serum metabolites. Cows with RFM had significantly higher concentrations of beta-hydroxybutyrate, non-esterified fatty acids and triglycerides after calving, but had lower concentrations of cholesterol during transition period. The concentrations of serum albumin and blood urea nitrogen were also significantly lower in RFM-affected cows than non-affected ones after parturition. Our results suggested that negative energy balance (NEB) postpartum was associated with RFM in dairy cattle. However, our findings did not reveal a cause and effect relationship with respect to the role of NEB as a possible risk factor for RFM.     

Post‐partum leucocytic activity and its relationship to caesarian section and retained placenta

Summary

The activity of the leucocytes of30 cows during 10 days after parturition is described. A comparison is made between the activity of leucocytes of 10 cows after normal parturition, 10 cows after caesarian section and 10 cows with retained placenta. When compared with normal parturition, caesarian section does not have a negative influence on the activity of the leucocytes during the puerperal period, during which time however the activity of leucocytes of cows with retained placenta is strongly decreased.     

Nitric oxide synthase expression in foetal placentas of cows with retained fetal membranes

The objectives of this study were to investigate relationship of retained fetal membranes (RFM) to expression of NOS and NOS mRNA and to analyze pathohistological changes and the distribution of nitric oxide synthase (NOS) in foetal placentas of cows with RFM. Twenty cows were assigned to two groups, a control group (no retained fetal membranes, NRFM, n = 10) and a diseased group (RFM, n = 10). The endpoint method was used to detect the nitric oxide (NO) content and nitric oxide synthase (NOS) activity in foetal placental tissue fluid and the fluorescent quantitation PCR was used to measure the expression of NOS mRNA. Immunohistochemistry and hematoxylin-eosin staining were used to observe pathohistological changes. Tissue from RFM cows showed fibronecrosis of the chorionic villi, and a decreased number of trophoblastic cells. The majority of trophoblastic cells displayed vacuolar degeneration. Interstitium vessels were distended and congested. Expression of induced nitric oxide synthase (iNOS) protein and iNOS mRNA was significantly higher (P < 0.05) in the cytoplasm of placental villus trophoblastic cells in the RFM group. But expression of endothelial nitric oxide synthase (eNOS) protein and eNOS mRNA was significantly lower (P<0.05) in the RFM group. The NO content and NOS activity of cows with RFM were significantly higher (P < 0.05). A high expression of iNOS protein and iNOS mRNA in the cow foetal placenta could produce high content of NO, which might inhibit uterine contraction. So over expression of iNOS protein and iNOS mRNA might be an important agent of retained fetal membranes in cows, and it may be a potential diagnosis biomarker.     

The effects of induced parturition on the incidence of clinical disease and mortality in dairy cows from commercial herds in south-western Victoria

SUMMARY The effects of induced parturition on clinical disease and mortality of cows were studied in 51 winter-calving, pasture-fed, dairy herds in south-western Victoria. Parturition was induced when most cows were between 27 and 35 weeks of pregnancy. Control groups were selected from herdmates that were not induced to calve. The incidence of retained placenta was much greater among induced cows of all ages. The proportion of cows affected by any clinical disease other than retained placenta, and the incidence of mortality were much greater after induction among cows aged over 6 years but not among cows aged 3 to 6 years.     

Type‐I interferon regulates matrix metalloproteinases clearance of the bovine endometrial spheroid

This study investigated the effect of type‐I interferon (IFN) on the expression of matrix metalloproteinases (MMPs) of the bovine endometrial stromal cells (BES) and epithelial cells (BEE). The cells were separated and purified from the caruncles and cultured in DMEM/F‐12 containing 10% fetal bovine serum. Spheroids were generated by using ascorbate. Zymograms of the supernatant showed that BEE predominantly expressed MMP‐9, whereas MMP‐2 was expressed in BES and homo‐spheroids. While MMPs expression was not detected in hetero‐spheroids. Real‐time quantitative PCR revealed that type‐I IFN and P4 suppressed the gene expression of MMP‐2 and MMP‐9 in hetero‐spheroids, respectively. On the other hand, gelatin zymography analysis of the supernatant showed that type‐I IFN strongly promote the clearance of MMPs. While zymograms of the MMPs stocked in the hetero‐spheroids were significantly reduced by type‐I IFN. Phenylmethanesulfonyl fluoride and leupeptin (both are serine proteinase inhibitors) significantly repressed the clearance of MMP‐2 and MMP‐9 induced by type‐I IFN. Moreover, collagen fibers in hetero‐spheroids significantly decreased after the treatment with type‐I IFN. In conclusion, it was suggested that type‐I IFN participate in the tissue remodeling by regulation the clearance of MMPs.     

Effect of clenbuterol administered during the expulsive stage of bovine parturition on uterine activity and the fetus.

The effect of an intravenous injection of 0.3 mg clenbuterol on myometrial activity, fetal heart rate and fetal outcome was studied in three groups of animals: six heifers in active labour at term, in which fetal oversize was diagnosed just before the drug was injected and whose calves were delivered by caesarean section within 50 minutes; four cows at the start of the expulsive stage of parturition which had been induced with flumethasone on day 270 of gestation; and in four parturient cows which had had electrodes implanted on the myometrium at least one week before calving was induced with flumethasone on day 270. Electrocardiograph electrodes were placed on the calf and an intrauterine pressure catheter was inserted between the calf and the uterine wall upon rupture of the amniotic membrane. Clenbuterol induced a significant decrease of myometrial activity for at least 20 minutes. Recovery was most rapid in the heifers in which an obstetrical examination had taken place during active labour. There was no significant effect on basal fetal heart rate but decreases in heart rate were absent as long as uterine contractions were inhibited. The fetal outcome, judged either before or at birth by blood pH, base-excess and PCO2 and by a clinical examination, was not adversely affected.     

Specific Localization of Macrophages in Pregnant Bovine Caruncles

The objective of this study was to evaluate the distribution and phagocytotic function of macrophages in bovine caruncles during pregnancy. Pregnant Holstein caruncles obtained at a slaughterhouse and by Caesarean section just before the onset of parturition were used. Macrophages in these caruncles were observed immunohistochemically and histochemically. Macrophages were present in the parenchyma of caruncles throughout pregnancy. The macrophages were irregular in shape, and exhibited thin, long processes along the main and intermediate septa of caruncles. Macrophages increased as the gestation period progressed. Acid phosphatase activity of the macrophages appeared at 5-6 months of gestation and remained until the end of gestation period. In particular, macrophages showed strong acid phosphatase activity at the peripheral region of the caruncles just facing the chorionic villi. In addition, they were conglomerated in the hyperplastic prominences of caruncles. These results indicate that caruncle macrophages maintain the appropriate size and shape of each caruncle during pregnancy.     

Effects of Hemophilus somnus on the pregnant bovine reproductive tract and conceptus following cervical infusion

Three strains of Hemophilus somnus were infused into the posterior cervix of six pregnant cows. The organism persisted in the cervicovaginal region for eight to 87 days, and at parturition H. somnus was isolated from chorioallantois in four of six cows; placentitis developed, and fetal membranes were retained. All calves were born alive and no H. somnus was recovered from them. One cow died 14 days after parturition. The death was attributed to severe necrotizing metritis; H. somnus was not isolated from the uterus at death, but was isolated from the placenta at parturition and cervicovaginal mucus two days days later.     

Glucocorticoid receptor down-regulation in neutrophils of periparturient cows

OBJECTIVE: To determine effects of parturition on glucocorticoid receptor (GR) expression in neutrophils, serum cortisol concentration, and total blood leukocyte and neutrophil counts in periparturient dairy cows. ANIMALS: 23 Holstein cows. PROCEDURE: Blood samples were collected from 8 multiparous and 5 primiparous periparturient cows at various times from 28 days before parturition until 14 days after parturition. Glucocorticoid receptor expression in neutrophils, serum cortisol concentration, and total blood leukocyte and neutrophil counts were determined. Results were compared with results from control samples obtained from 5 multiparous and 5 primiparous Holstein cows in midpregnancy. RESULTS: Neutrophils from periparturient cows had 49% reduction in GR expression at calving, compared with GR expression 2 to 4 weeks before calving, and 39% reduction, compared with neutrophils from cows in midpregnancy. Reduction in neutrophil GR expression began 1 week before calving and was most severe at calving and 24 hours after calving; a significant difference in GR expression was detected between primiparous and multiparous cows. Serum cortisol concentrations and total leukocyte and neutrophil counts were significantly increased at calving and returned to baseline values by 24 hours after calving. Significant negative correlations were detected between neutrophil GR expression and serum cortisol concentration, total leukocyte count, and neutrophil count. CONCLUSIONS AND CLINICAL RELEVANCE: Reduced GR expression in blood neutrophils of periparturient dairy cows was associated with increased serum cortisol concentrations, leukocytosis, and neutrophilia. Thus, GR down-regulation in neutrophils may be involved in periparturient neutrophil dysregulation and may cause increased susceptibility to mastitis.     

The effects of treatment with cloprostenol or dinoprost within one hour of induced parturition on the incidence of retained placenta in cattle

Two experiments were designed to determine whether prostaglandin treatment within one hour postpartum would reduce the incidence of retained placentas after induction of parturition in beef cattle. In the first experiment, 70 cows were induced on day 276-278 of gestation with the combination of 500 μg cloprostenol and 25 mg dexamethasone (CP + Dex). Within one hour after parturition, cows received either 500 μg CP or 25 mg of dinoprost (DI). The incidence of retained placenta (RP) was 64.3% in induced groups and 0% in noninduced control cows and postpartum treatment with either CP or DI had no effect on placental retention.

A second experiment, utilizing 132 cows and heifers, was conducted to determine whether induction with Dex alone, rather than with CP + Dex, would influence the rate of placental retention after postpartum treatment with either CP or DI. The incidence of retained placenta ranged from 28.5 to 58.3% in induced females but was 0% in noninduced control females. As in the first experiment, postpartum prostaglandin treatment had no effect on placental retention.

The results of these experiments do not support the use of prostaglandins within one hour of induced parturition to reduce the incidence of retained placentas.

     

Physiology and Treatment of Retained Fetal Membranes in Cattle

Retained fetal membranes (RFM) in cattle have adverse effects on fertility and production. Understanding the pathophysiology and causes of RFM is important for managing this disease. The hormonal processes that lead to normal placental separation are multifactorial and begin before parturition. A variety of risk factors, including early or induced parturition, dystocia, hormonal imbalances, and immunosuppression, can interrupt these normal processes and result in retention of the placenta. Current research does not support the efficacy of many commonly practiced treatments for RFM. Systemic administration of antibiotics can be beneficial for treating metritis after RFM, but antibiotic administration has not been shown to significantly improve future reproduction in cows with RFM. Collagenase injected into the umbilical arteries of retained placentas specifically targets the lack of placentome proteolysis and might enhance placental release. However, such therapy is costly and its benefits in terms of improving subsequent reproductive function have not been evaluated.