Patterns of protein glycosylation in bovine placentomes as a function of gestational age and in retained versus non‐retained placenta

The formation of placenta at the beginning of pregnancy and its separation at parturition require not only deep remodelling of extracellular matrix, which mainly consists of proteins conjugated with sugar moieties, but also the cooperation with cells from both maternal and foetal parts of placenta. The aim of the study was to compare the patterns of selected conjugated proteins with sugar moieties between pregnant and term placenta as well as between released and retained placenta in cows. Placental samples from healthy pregnant cows (3–5 months of pregnancy) were collected at a slaughterhouse (n = 6), and parturient samples were collected during caesarean section at term and retrospectively divided into retained (n = 6) and released (n = 6). The pattern of selected sugar moieties conjugated with proteins was detected by use of lectin blotting with Phaseolus Vulgaris leucoagglutinin, Maackia Amurensis and Sambucus Nigra (Elderberry). The comparison and analysis of obtained band patterns showed differences between their number, molecular weight and abundance related to the intensity of staining. Samples from 3 to 4 months showed similarities, while at the 5th month, clear differences were visible in all 3 lectins, which were used in this study. Samples from retained/released placenta expressed significant differences in PHA‐L and SNA pattern in the foetal part. Obtained results indicate that the development of placenta related to extracellular matrix and accompanying cells from both sides of placenta shows dynamic changes during pregnancy. Moreover, in the case of animals with the retention of foetal membranes the patterns of proteins conjugated with sugar moieties are altered, suggesting that the changes in extracellular matrix metabolism can be involved in the attachment and detachment of the placenta in cows.     

Poly(ADP-Ribose) Glycohydrolase in Bovine Retained and Not Retained Placenta

Poly(ADP-ribose) glycohydrolase (PARG) is the enzyme which degrades poly(ADP-ribose) polymers synthesized by poly(ADP-ribose) polymerase (PARP). Both enzymes are activated in response to different stimuli like oxidative stress and are involved in DNA repair processes. The retention of bovine foetal membranes (RFM) is supposed to be connected with oxidative stress conditions. The aim of the study was to detect the presence of PARG protein in bovine placenta in order to find the relationship between the process of releasing, retaining placenta and DNA repair. Placentomes, collected alter spontaneous delivery or caesarian section were divided into maternal as well as foetal part of placenta, homogenized and subjected to electrophoresis. Animals were divided into six groups as follows: A--caesarian section before term with RFM; B--caesarian section before term without RFM; C--spontaneous delivery at term with RFM; D--spontaneous delivery at term without RFM; E--caesarian section at term with RFM; F--caesarian section at term without RFM. PARG protein was detected in nitrocellulose membranes using commercially available bovine anti-PARG antibody and Western blotting technique. Single bands referred to bovine PARG standard were observed in all examined tissues as well as in human placenta used as the control of procedure. In addition, the intensity of staining was stronger in retained than properly released term placenta and in foetal than in maternal part of the placenta. These results may suggest the differences in enzyme protein content and careful conclusions can be drawn that the activities of PARG may be altered between compared groups of animals. It may confirm the presence of oxidative stress conditions and their consequences on metabolic pathways, the content of biologically active substances and processes of proper releasing placenta. Further experiments on PARG activity in bovine foetal membranes with respect to proper and improper placental release are necessary.     

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 effect of chloride ammonium, vitamin E and Se supplementation throughout the dry period on the prevention of retained fetal membranes, reproductive performance and milk yield of dairy cows

We assessed the effect of an anionic salt and supplementary administration of vitamin E and Se throughout the dry period, on the risk of retained fetal membranes (RFM), milk yield and reproductive performance of dairy cows. Data were collected from 456 dairy cows in three commercial farms. Each animal entering the dry period was assigned to one of two groups (treated and control group). All animals were then fed the same ration, which included 80 IU/kg vitamin E acetate and 0.2 ppm Se, but animals of the treated group also received an additional blend containing ammonium chloride (60 g), vitamin E [1000 IU (dl-α-tocopheryl acetate)] and Se (0.05 ppm). Calving ease was evaluated and no manual removal of placenta was attempted. Cows that retained their fetal membranes for more than 12 h after calving were considered to suffer from RFM. All animals experienced a 50-day voluntary waiting period before the first artificial insemination (AI). Treatment resulted in a decrease in the estimated average risk of RFM (10.6% vs 17.8%). Stratifying on farm the Mantel–Haenszel Relative Risk (MH-RR) estimate of RFM in treated animals relative to controls was 1.68 (95% confidence interval, CI, 1.05 to 2.68) and the Mantel–Haenszel test P-value was 0.028. Among cows that had not required assistance during delivery the average risk of RFM was greater for those without treatment (risk estimate: 10/208 or 9.7%) as compared to those with treatment (risk estimate: 20/207 or 4.8%). The MH-RR estimate of RFM in treated vs. control animals was 2.95 the respective 95% CI: (0.96 to 4.17) and the MH test of association P-value was 0.058). Conversely, among cows that did require assistance during calving, the average risk of RFM estimates were 77.8% (14/18) and 91.3% (21/23) in treated and non-treated animals, respectively (MH-relative risk estimate of RFM = 1.21, 95%CI: 0.91 to 1.60). Using analysis of variance with farm as a random effect, treatment did not appear to have an effect on milk production of cows without RFM either at 30 or 60 days postpartum (P > 0.10). Time intervals (in days) between parturition and first oestrus expression (POI), parturition and first AI (PFAI) and parturition and conception (PCI) were recorded for each animal. For each time-to-event variable, survivor functions were estimated for treated and control groups using the Kaplan–Meier methodology and survival curves of treated and control groups were compared using the log-rank test, separately for the animals that did or did not experience RFM in each farm. There were significant differences in the time-to-event survival curves in only one of the three farms. Median time between parturition and first oestrus expression among animals without RFM was 67 days in treated animals and 75 in control animals, and the survival curves were statistically significantly different (P = 0.021). Similarly, in the same farm, among animals that did not experience RFM the median time between parturition and conception was 114 in treated animals and 145 in controls, and the survival curves were statistically significantly different (P = 0.002). In conclusion, daily administration of a blend containing ammonium chloride, vitamin E and Se throughout the dry period seems to be safe and resulted in a decrease of RFM occurrence, without any effect on milk yield.     

15-Ketodihydro-PGF2α, Progesterone and Uterine Involution in Primiparous Cows with Induced Retained Placenta and Post-partal Endometritis Treated with Oxytetracycline and Flunixin

Retention of the foetal membranes (RFM) and post‐partal endometritis are common problems in dairy cows. Among other things, the disease is characterized by a bacterial endometritis with aerobic as well as anaerobic bacteria. From an endocrine perspective, cows with RFM have high levels of 15‐ketodihydro‐PGF_2α (PG‐metabolite) immediately after parturition but these levels fall rapidly within 2 weeks post‐partum (early PG‐metabolite elevation). After this decline, the PG‐metabolite levels increase again and the levels (at this time of a lower magnitude) remain elevated during the period of uterine infection (late PG‐metabolite elevation). The aim of this study was to investigate the PG‐metabolite profiles in cows with retained placenta and post‐partal endometritis treated with the prostaglandin synthesis inhibitor flunixin (F), either alone or in combination with oxytetracycline (T). The study was accomplished over 2 years with 12 primiparous cows in each experiment. As a model for RFM, preterm parturition was induced in late‐pregnant heifers by injecting PGF_2α (25 mg i.m) twice with a 24 h interval. In each experiment, the cows were divided into four groups and treated with either T (10 mg/kg b.w. i.m. once daily), F (2.2 mg/kg b.w. p.o. twice per day), a combination of T and F (dosage, as above), or conservatively (0). The treatment periods lasted from day 11 to day 14 post‐partum (pp) in experiment 1 (after placental shedding, groups T1, F1, TF1 and 0) and from day 3 to day 6 pp in experiment 2 (before placental shedding, groups T2, F2, TF2 and 0). Jugular vein blood samples were collected for analyses of PG‐metabolite and flunixin. Uterine biopsies were collected twice weekly for investigation of endometrial microbiology. Rectal palpation and ultrasonographic examinations were performed three times per week for investigations of uterine and cervical involution and ovarian activity. No attempts were made to remove the placentas manually. The experiment lasted until day 56 pp. The induction of parturition was successful in all heifers and 22 of 24 animals had RFM. All RFM cows had bacterial endometritis, based on bacteriological examinations. Flunixin treatment (F1, TF1, F2 and TF2) suppressed PG‐metabolite levels significantly (p=0.006) during the period of treatment in both experiments. However, the early flunixin treatment only suppressed PG synthesis partially. Late oxytetracycline treatment (T1) did not influence the PG‐metabolite levels but oxytetracycline treatment (T2 and TF2) before placental shedding significantly altered the kinetics of the early PG‐metabolite elevation compared with other treatments. Late PG‐metabolite elevation was significantly correlated to duration of uterine infection and cervical involution. In conclusion, flunixin treatment of cows with retained placenta either before or after placental shedding suppresses prostaglandin synthesis. However, early treatment, when the release of prostaglandins is high, might need more intensive treatment in order to prevent the PG synthesis effectively. Oxytetracycline treatment during the period immediately after parturition before placental shedding might influence the PG‐metabolite profile and suggests a bacteriological contribution to the high levels of PG‐metabolite seen during the first 2 weeks pp in cows with retained placenta. In this study, a correlation between prostaglandin release, the final cervical involution and the end of infection was found. This suggests a link between uterine endocrinology, bacteriology and involution in cows with retained placenta.     

Bacterial Isolates Associated With Dystocia And Retained Placenta In Iraqi Buffaloes

The present study was conducted on 50 recently calved Iraqi Buffalo cows. Depending on the kind of parturition, buffalo cows were divided into two main groups, the first group had normal unassisted parturition (NP) (26 animals) and the second group with certain periparturent complications (PPC) (24 animals). After 24 h of parturition, these two groups were further subdivided into two groups as cows expel their foetal membranes in <24 h postpartum and referred as non‐retained placenta (NRP) while cows that did not expel their foetal membrane after 24 h referred as retained placenta (RP). Sampling for bacteriology, uterine discharge for polymorphonuclear cells per cent and blood samples for polymorphonuclear neutrophil (PMN) and the enzyme creatine kinase activity were performed at 6, 24 and 48 h postpartum. In PPC group, the most prevalent bacteria after 6 h of calving were Escherichia coli, β‐haemolytic Streptococci and Lactobacillus acidophilus. Total bacterial isolates in the uterus of buffaloes with RP in PPC group after 24 and 48 h were 129 and 183 respectively. Among the isolates, Archanobacterium pyogenes, Fusobacterium necrophorum, Prevotella melaninogenicus and Staphylococcus aureus were the most prevalent isolates after 48 h of RP buffaloes in PPC group. Polymorphonuclear neutrophil were significantly (p < 0.01) increased in the uterine discharge than in blood in buffaloes with RP in both PPC and NP groups. In conclusion, uterine contamination occurs as a result of postpartum ascending contamination by non‐specific environmental organisms. The presence of Lactobacillus sp. in the uterus indicated a healthy uterus. Peripartum complications followed by retention of foetal membranes with the dominance of E. coli in the uterine lumen might favour the colonization of other bacteria including facultative anaerobic and strictly anaerobic in the uterine wall of buffaloes.     

Clinical and Bacteriological Aspects on the Use of Oxytetracycline and Flunixin in Primiparous Cows with Induced Retained Placenta and Post-partal Endometritis

Retention of the fetal membranes and post‐partal endometritis (RFM) are common problems in dairy cows. Treatment often includes manual removal of the placenta in combination with antibiotic treatment. Earlier studies have shown that cows with endometritis post‐partum have a strong tendency to recover spontaneously. The present study focused on treatments of post‐partal endometritis with the prostaglandin synthesis inhibitor, flunixin (F) either alone or combined with oxytetracycline (T). The study was conducted in two experiments, using 12 primiparous cows in each. As a model for RFM, premature parturition was induced in late pregnant heifers by injecting PGF_2α (25 mg i.m.) twice with a 24 h interval. In each experiment the cows were set into four groups and treated with either T (10 mg/kg BW i.m. once daily), F (2.2 mg/kg BW p.o. twice daily), a combination of T and F (dosage, as above) or conservatively (group 0, no drugs). The treatment periods lasted from days 11–14 post‐partum in experiment 1 (groups T1, F1, TF1 and 0) and from days 3–6 post‐partum in experiment 2 (groups T2, F2, TF2 and 0). Jugular vein blood samples were collected for analyses of flunixin and total white blood cells. Uterine biopsies were collected twice weekly for investigation of endometrial microbiology. Rectal palpation and ultrasonographic examinations were performed three times weekly for investigations of uterine involution and ovarian activity. No attempts were made to remove the placentas manually. The experiment lasted until day 56 post‐partum. The induction of parturition was successful in all heifers and 22 of 24 animals had RFM. All RFM cows had bacterial endometritis. The predominant bacteria were Escherichia coliα‐haemolytic streptococci, Fusobacterium necrophorum, Arcanobacterium (Actinomyces) pyogenes, Bacteroides spp., Pasteurella spp. and Proteus spp. Fusobacterium necrophorum and A. pyogenes could be isolated for 3–5 weeks post‐partum and E. coli, Pasteurella and Proteus could be isolated for 2–3 weeks post‐partum. Animals treated with tetracycline after placental shedding (T1 and TF1) had a more rapid recovery from infections with A. pyogenes and F. necrophorum than animals that were not treated with tetracycline. No other genera were affected. Antibiotic treatment before placental shedding (T2 and TF2) did not shorten the uterine infection but altered the bacterial flora, seen as an overgrowth of Proteus spp. (p < 0.05) and increased frequency of Pasteurella (p < 0.05). The α‐haemolytic streptococci were less common in T2 and TF2 than in other groups (NS). Antibiotic treatment of cows before placental shedding (T2 or TF2, n=6) postponed detachment of placenta compared to cows were no antibiotics were administered before placental shedding (T1, TF1, F1, F2 and 0, n=16, 9.8 days pp (median) versus 11.8, p=0.004). Neither treatment shortened uterine involution. Flunixin treatments did not seem to influence recovery from infection or uterine involution. It was concluded that early oxytetracycline treatment of retained fetal membranes in the cow did not shorten the uterine involution or uterine infection but it did slow down the detachment process of the retained placenta. Oxytetracycline treatment after placental shedding might shorten the uterine infection but otherwise did not affect the clinical results. Flunixin treatment had no influence on the clinical outcome of the disease.     

Changes of leukocyte counts and expression of pro‐ and anti‐inflammatory cytokines in peripheral leukocytes in periparturient dairy cows with retained fetal membranes

In dairy cows, retained fetal membranes (RFM) affect reproductive performance. The aim of this study was to examine the leukocyte counts and the gene expression of tumour necrosis factor α (TNFα), interleukin 1β (IL‐1β), IL‐8, and IL‐10 in polymorphonuclear leukocytes (PMNs) and peripheral blood mononuclear cells (PBMCs) in cows with (n = 5) or without (n = 5) RFM during the peripartum period. The lymphocyte counts in RFM cows were higher than those in control cows throughout the experiment (p < .05). The expression of IL‐8 in PMNs of control cows was higher (p < .05) compared with that of RFM cows postpartum. In cows with RFM, IL‐1β expression was higher (p < .05) in PMNs at 6 weeks postpartum whereas the expression of IL‐1β was lower (p < .05) in PBMCs at 4 weeks postpartum. The expression of IL‐10 in PBMCs of control cows was higher (p < .05) than that of RFM cows at 2 weeks prepartum and 4 weeks postpartum. Taken together, our data indicate that changes of gene expression of pro‐ and anti‐inflammatory cytokines in RFM cows might be associated with the delayed placental separation and development of uterine inflammation in RFM cows.     

Activity of selected glycosidases and availability of their substrates in bovine placenta during pregnancy and parturition with and without retained foetal membranes

The activity of glycosidases is crucial for the function and biological activity of proteins conjugated with sugar moieties, which play an important role in adhesion of cells during attachment and detachment of the foetal membranes. The aim of study was to describe the ability of bovine placental tissues to break down O-glycosidic bonds in different glycoproteins by the determination of activity of β-galactosidase, α-l-fucosidase, β-N-acetyl-hexosaminidase and sialidase in early–mid-pregnancy as well as at parturition with released and retained foetal membranes. Moreover, the availability of substrates for these glycosidases in placental homogenates was evaluated. Placental samples were collected from pregnant (2–4 months) cows in slaughterhouse (n = 8) as well as during Caesarean section and divided into released foetal membranes (n = 8) and retained foetal membranes (n = 8). Tissue homogenates were subjected to spectrofluorimetric and spectrophotometric determinations of enzyme activities as well as electrophoretic separations. Enzyme activities expressed changes within examined time with significant (p < .05) differences between pregnancy and physiological parturition in β-N-acetyl-hexosaminidase and α-l-fucosidase in foetal part of placenta while in maternal part only in the latter one. Decreasing tendency in enzyme activity was noticed in foetal part of retained samples in comparison with released ones with significant (p < .05) differences in α-l-fucosidase activity. The analysis of staining of sugar moieties attached to selected proteins depicted availability of sugar molecules in examined tissues, but their patterns differed between samples. In conclusion, sugar moieties in conjugated proteins express changes in the course of pregnancy which is reflected by the alterations in activities of placental glycosidases.     

Flutamide Influences Placental Aldo–Keto Reductase Family 1 Member C1 (AKR1C1) Expression in Pigs

Aldo–keto reductase family 1 member C1 (AKR1C1) catalyses the conversion of progesterone into inactive 20α‐dihydroxyprogesterone. It is suggested that AKR1C1 expression in the placenta prevents from the cytotoxic effect of progesterone on foetuses during late pregnancy. The aim of the study was to determine whether the anti‐androgen flutamide administered during late pregnancy (83–89 days of gestation) or before parturition (101–107 days of gestation) influences AKR1C1 expression in the porcine placenta. AKR1C1 mRNA and protein levels were measured using real‐time PCR and western blotting, respectively. Immunolocalization of AKR1C1 within placentas was also performed. Flutamide significantly increased AKR1C1 mRNA (p = 0.008) and protein (p = 0.019) expression only during the pre‐parturient period in pigs. AKR1C1 protein was immunolocalized in the epithelial and stromal cells of foetal and maternal part of placenta at both stages of gestation. Following flutamide treatment, the intensity of staining was higher (p = 0.045) on day 108 of gestation. In conclusion, porcine placental AKR1C1 expression seems to be regulated by an androgen signalling pathway and may be involved in foetal survival by preventing the detrimental effect of progesterone.     

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.     

Enzymatic activity in the placenta in cows in relation to the course of parturition

The activity of aminopeptidases and cathepsins was determined in placentoma homogenates; placentomas of cows were extirpated immediately after parturition and in four and eight hours. In cows with afterbirth retention (a. r.) following induced parturition, the activity of these enzymes was always higher than in cows without a. r., no matter if after induced and spontaneous parturitions; it was at a similar or slightly higher level than in the eighth month of pregnancy. The content of total proteins in placentomas of cows with a. r. was also higher. These findings point to the insufficient ripening, or aging of placental tissue, which is related to a release of lysosomal enzymes. The enzymes are active through hydrolysis in the separation of foetal placenta from the maternal one and in the modification of proteins participating in intercellular linkages.     

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.     

Analysis and Validation of the Differentially Expressed ENO1 in the Maternal Placenta of Cow with Retained Foetal Membrane

The paper aimed to analyze and validate the function of differentially expressed ENO1 in the cow with retained foetal membrane.In our research,we chose three healthy Holstein dairy cows and three Holstein dairy cows with retained foetal membrane of similar age,foetal times,weight and milk yield and divided them into two groups.The total protein of maternal placenta was extracted in the control and retained foetal membrane of cow.The differential expression of proteins were found out by the 2-DIGE,and the differential expression of ENO1 was validated by the Western blotting and Real-time quantitative PCR.The results showed that ENO1 was significant expression and large multiple in the two groups,and it was significant increased expression in the retained foetal membrane of cow after Western blotting and Real-time quantitative PCR (P=0.015<0.05;P=0.001<0.01).The ENO1 participated in the energy homeostasis,immune and fibrinolysis process which related to the retained foetal membrane of cow.It suggested that ENO1 was likely connected with the retained foetal membrane.     

胎衣不下奶牛母体胎盘α-烯醇化酶异常表达分析及验证

试验通过对胎衣不下奶牛母体胎盘组织中差异α-烯醇化酶(enolase,ENO1)的分析验证,探讨ENO1在胎衣不下中的作用。本研究选取了年龄、胎次、体重和泌乳量均相近的产后胎衣不下和产后胎衣正常排出奶牛各3头,分为两组,提取了胎衣不下组与胎衣正常排出组母体胎盘组织中的总蛋白,采用双向凝胶电泳的方法筛选出差异蛋白,并利用Western blotting及实时荧光定量PCR的方法对其中差异表达量大的ENO1进行验证。结果发现,胎衣不下组和胎衣正常组母体胎盘组织中ENO1差异表达量大且倍数较大,Western blotting及实时荧光定量PCR验证发现ENO1在胎衣不下奶牛母体胎盘中的表达量升高,t检验结果分别为P=0.015<0.05,差异显著;P=0.001<0.01,差异极显著。该基因参与机体的能量调节、免疫和纤溶等过程,而这些过程与奶牛胎衣不下密切相关,提示ENO1可能参与该病的发生发展。     

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.     

Nitrate poisoning in cattle

Summary

In a series of experiments the effect of administering KNO₂ was studied, during parturition, on the capability of oxygen transport of maternal blood and on oxygen transfer to foetal blood. The following blood parameters were analysed, MHb percentage, pO₂, O₂‐saturation, pH, pCO₂, and (NO₂) in maternal arterial blood (carotid art.) and venous blood (jugular vein) and in foetal arterial blood (umbilical art.) and venous blood (umbilical vein). The relative O₂‐saturation was calculated from the estimated O₂‐saturation by multiplying with the factor Hb (mmol/1) minus MHb (mmol/1), divided by Hb (mmol/1). In addition, blood pressure in the carotid artery, heart rate, and respiration rate in the dam were continuously recorded for some hours.

A dosage of 9 to 12 mg of NO₂/kg body weight intravenously or of 30 mg of NO₂/kg body weight orally to the dam caused much higher MHb percentages and NO₂ contents in the maternal blood than in the foetal blood. In maternal blood the ratio of NO₂ content td MHb percentage was proportional to that in foetal blood. In the arterial blood, MHb percentages were almost as high as in the venous blood. After administering of nitrite, relative O₂‐saturation dropped simultaneously with the increase in methaemoglobin.

Nitrite treatment caused a drop in the maternal blood pressure; heart rate and respiration rate increased. O₂‐saturation in the blood in the umbilical vein was much lower in the animals with nitrite treatment than in those without. These experimental results show clearly that the oxygen capacity of the blood decreases after nitrite treatment. In pregnant cows the oxygen supply to the foetus will be adversely affected after nitrate intake, especially by the lower oxygen transfer via the placenta, though hardly at all by methaemoglobin formation in the foetal blood.

When the oxygen transfer to the foetal blood decreases too sharply, intra‐uterine death and ultimately abortion may result.     

Foetal development of skeletal muscle in bovines as a function of maternal nutrition,foetal sex and gestational age

To determine the effects of maternal nutrition on modifications of foetal development of the skeletal muscle and possible increase in the potential of skeletal muscle growth in cattle, gestating cows were either fed 190% NRC recommendations (overnourished; ON) or 100% NRC recommendation (control; CO). Interaction between maternal nutrition (MN) and the foetal sex (FS) was also investigated. Foetuses were necropsied at four different time points throughout gestation (139, 199, 241 and 268 days of gestation) to assess the mRNA expression of myogenic, adipogenic and fibrogenic markers in skeletal muscle. Phenotypic indicators of the development of skeletal muscle fibres, intramuscular lipogenesis and collagen development were also evaluated. Modifications in mRNA expression of skeletal muscle of foetuses were observed in function of MN and FS despite the lack of effect of MN and FS on foetal weight at necropsy. Maternal ON increased the mRNA expression of the myogenic marker Cadherin‐associated protein, beta 1 (CTNNB1) and adipogenic markers Peroxissome proliferator‐activated receptor gamma (PPARG) and Zinc finger protein 423 (ZNF423) at midgestation. However, no differences on foetal skeletal muscle development were observed between treatments at late gestation indicating that a compensatory development may have occurred on CO foetuses making the effect of MN on skeletal muscle development not significant at late gestation. Moreover, our data have shown an evidence of sexual dimorphism during foetal stage with a greater skeletal muscle development in male than in female foetuses. In conclusion, providing a higher nutritional level to pregnant cows changes the trajectory of the development of skeletal muscle during midgestation, but apparently does not change the potential of post‐natal growth of muscle mass of the offspring, as no differences in skeletal muscle development were observed in late gestation.     

Effects of periparturient diseases and conditions on the reproductive performance of New Zealand dairy cows

AIM: To determine the effect of retained foetal membranes (RFM), periparturient recumbency, calf mortality, dystocia, twin births and induction of parturition on submission and pregnancy rates of pasture-fed, seasonally-calving dairy cows in New Zealand. METHODS: Data were retrospectively collected for 2652 cows from 11 herds on the occurrence of periparturient diseases and condition, calving date, age and treatment for anoestrus prior to the planned start of mating (PSM). The effects of each disease or condition on submission and pregnancy rate by 28 days after PSM, final pregnancy rate, and the proportion of cows that conceived more than 50 days after PSM were examined using separate logistic regression models. Results are expressed as odds ratios (OR). Additionally, the effect of each disease or condition on the interval from PSM to conception was examined using survival analysis. Age, herd and late calving (i.e. cows calved < or =40 days before PSM) were included as factors in the models examined. RESULTS: Induction of parturition increased the risk of RFM (OR=3.3, p<0.01). The 28-day submission rate was affected by age and herd but not by any of the periparturient conditions examined. The 28-day pregnancy rate was reduced by RFM (OR=0.04, p<0.05), induction of parturition (OR=0.67, p<0.01), assisted calving (OR=0.61, p<0.01), late calving (OR=0.55, p<0.05) and anoestrus (OR=0.27, p<0.05). The final pregnancy rate was reduced by RFM (OR=0.013, p<0.01), induction (OR=0.25, p<0.05), assisted calving (OR=0.30, p<0.005) and anoestrus (OR=0.32, p<0.05), and was increased if the cow was submitted for mating within 28 days after PSM (OR=2.72, p<0.05). Effects of recumbency and twinning were not significant in any of the models. CONCLUSIONS: Cows that had had RFM or were assisted, induced or late to calve, had significantly lower pregnancy rates or took longer to conceive than unaffected herd-mates. CLINICAL RELEVANCE: Intervention programs aimed at the examination and treatment of cows with periparturient conditions may improve the reproductive performance of New Zealand dairy herds. Investigations of poor reproductive performance of a herd should include analysis of the prevalence and effects of periparturient diseases and conditions.