Leukotriene B4 in cows with normal calving, and in cows with retained fetal membranes and/or uterine subinvolution.

Two experiments were performed to study the relationship between leukotriene B4 (LTB4) synthesis and placental separation and uterine involution in the cow. In experiment I, the concentration and synthesis of LTB4 by caruncular tissue was lower in cows with retained fetal membranes (RFM cows, n = 11) than in cows that expelled the fetal membranes normally (NFM cows, n = 19). The presence of bacterial cell wall, especially of alpha-hemolytic streptococci and coagulase positive staphylococci enhanced LTB4 synthesis by allantochorion only in NFM cows. In the RFM group, Escherichia coli lipopolysaccharide decreased allantochorionic LTB4 synthesis. With caruncle, only epidermal growth factor increased LTB4 production in NFM cows. In experiment II, the caruncular and endometrial secretion of LTB4 was lower in cows with subuterine involution (SUI cows, n = 5) or cows with SUI and RFM (SUI+RFM cows, n = 4) than in cows with normal uterine involution (NUI cows, n = 8). This decrease was especially noticeable in the previously gravid horn. In the three uterine involution groups, there were no differences in LTB4 synthesis by caruncular tissue taken from the previously gravid horn. However, progesterone and a bacterial suspension of E. coli reduced the synthesis of LTB4. Estradiol had no effect on LTB4 synthesis at the end of the postpartum period. These results suggest that LTB4 may play an important role in both placental separation and uterine involution in cattle and LTB4 synthesis may be modulated by endocrine and bacterial factors.     

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.     

Aspects of bacteriology and endocrinology of cows with pyometra and retained fetal membranes

In an effort to clarify the pathogenesis of pyometra, 20 cows with retained fetal membranes and 20 without, but with contemporary calving dates were studied. They were palpated and their uteri were subjected to sample collections for bacteriologic cultural examinations twice weekly for 4 weeks. Blood samples were obtained each day and evaluated for serum progesterone concentration. Three cows without and 3 with retained fetal membranes developed pyometra during the study, resulting in 3 groups designated control (CON), cows with retained fetal membranes (RFM), and cows with pyometra (PYO). Bacterial isolations occurred less frequently in the CON group than in the PYO and RFM groups. Growth patterns of bacteria also varied between groups. Coliform and incidental bacteria disappeared from the uterus of the PYO group by the end of the 3rd week. In contrast, heavy growth of Corynebacterium pyogenes and gram-negative anaerobic bacteria developed during this same period in the PYO group. In cows with pyometra, the significant persistent pathogenic bacteria recovered were C pyogenes and gram-negative anaerobic bacteria, especially Fusobacterium necrophorum and Bacteroides melaninogenicus. Anaerobic bacteria were isolated simultaneously with C pyogenes in most cows of the PYO group, but less often in CON and RFM groups, and highest growth levels were present near the time of ovulation. Clinically, pyometra usually developed about 10 days after observation of concurrent ovulation and high growth levels of C pyogenes and gram-negative anaerobic bacteria. A hypothesis is presented for development of pyometra in the cow.     

Evaluation of the effects of treating dairy cows with meloxicam at calving on retained fetal membranes risk

Some non-steroidal anti-inflammatory drugs increase the risk of retained fetal membranes. This is the first study to investigate the effects of meloxicam on the risk of retained fetal membranes. Administration of meloxicam to dairy cattle immediately following calving revealed no differences in the incidence of retained fetal membranes between meloxicam-treated and untreated animals. There was no difference between the 2 groups in the incidence of periparturient diseases following calving. Meloxicam can be used on the day of calving in lactating cows without increasing the risk of retained fetal membranes.     

Effect of intrauterine administration of ceftiofur on fertility and risk of culling in postparturient cows with retained fetal membranes, twins, or both

OBJECTIVE: To determine the effect of intrauterine administration of ceftiofur sodium on fertility and the risk of culling in postparturient cows with retained fetal membranes (RFM), twins, or both. DESIGN: Single-blind randomized clinical trial and prospective cohort study. ANIMALS: 2442 cows that calved from January 1, 2000, to May 31, 2001. PROCEDURE: Cows with RFM, twins, or both were randomly allocated to control or treatment (ceftiofur) groups. Ceftiofur-group cows received 1 g of ceftiofur sodium sterile powder reconstituted with 20 mL of sterile water as a single intrauterine infusion once between 14 and 20 days after parturition. Control-group cows received no treatment. Cows that calved but did not have RFM or twins were considered the referent group. Reproductive, culling, and health data were recorded. RESULTS: There was no significant difference in the overall proportion of ceftiofur-group cows confirmed pregnant, compared with cows in the control group. Ceftiofur-group cows were significantly less likely to be culled and were culled at a later time in lactation than control-group cows. In the cohort study, the risk of pregnancy and the risk of being culled in ceftiofur-group cows were not significantly different from cows in the referent group. CONCLUSIONS AND CLINICAL RELEVANCE: Intrauterine treatment of cows with RFM, twins, or both with ceftiofur sodium increased longevity of cows in the herd as measured by the risk of culling and the time to culling. Intrauterine administration of ceftiofur in cattle is considered extralabel drug use, and the attending veterinarian must follow the AMDUCA guidelines for extralabel drug use.     

酒炙车前子防治奶牛胎衣不下中有效物质分离体系的确定

酒炙车前子防治奶牛胎衣不下是验方,本实验室验证酒炙车前子在防治奶牛胎衣不下方面有着显著的疗效,并确定了有效部位。但其防治奶牛胎衣不下的有效物质基础尚不明确。因此本试验目的是确定一个能够将其有效部位中所包括的化合物按照极性由低到高分离成三部分的体系。通过预实验确定了分离二氯甲烷部位和乙酸乙酯部位的洗脱剂和展开剂体系。使用相同内径的层析柱,其中硅胶的高度设置成3个梯度:15,25,35cm,使用洗脱剂进行梯度洗脱后,利用薄层色谱法进行检测,由色谱图观察化合物是否能成功分离成三部分。二氯甲烷部位和乙酸乙酯部位的结果显示,柱高15cm条件下无法有效地分离化合物,柱高25cm条件下化合物回收比例要高于柱高35cm条件下的化合物回收比例。因此确定了二氯甲烷部位分离化合物的洗脱剂体系:石油醚∶乙酸乙酯为1∶2;石油醚∶乙酸乙酯为1∶30;二氯甲烷∶甲醇为30∶1;展开剂体系:石油醚∶乙酸乙酯为4∶1;柱体高度为25cm。乙酸乙酯部位分离化合物的洗脱剂体系:石油醚∶乙酸乙酯为100∶1;石油醚∶乙酸乙酯30∶1;二氯甲烷∶甲醇为30∶1;展开剂体系:石油醚∶乙酸乙酯为10∶1;柱体高度为25cm。采用干法上样,上样量应适当增加。该分离体系的确定对研究酒炙车前子中防治奶牛胎衣不下的有效物质基础提供了技术支撑,也为研制高效、安全、廉价的防治奶牛胎衣不下的中药制剂奠定了基础。     

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.     

围产期酮病奶牛血液生化指标特征分析

试验随机选取产前奶牛29头,跟踪采集产前2周、产前1周、分娩当天及产后3,7,14,21 d的血浆样品,检测血浆相关生化指标变化特征,并记录泌乳量。任意采集时间点血浆β-羟丁酸(BHBA)≥1.2 mmol/L则为酮病组, BHBA<1.2 mmol/L则为对照组。结果显示:随机选取的牛群中酮病发病率为58.6%,酮病主要发生在产后7～14 d;酮病组奶牛产前血液相关生化指标与对照组相比无显著性差异;与对照组相比,酮病组奶牛分娩当天血钾水平升高,血磷水平显著降低;产后3 d酮病组奶牛血浆甘油三酯、总胆固醇(TC)、高密度脂蛋白胆固醇(HDL-C)、BHBA、泌乳量显著高于对照组奶牛;产后7,14,21 d血浆BHBA、非脂化脂肪酸(ENFA)、TC、HDL-C、低密度脂蛋白胆固醇(LDL-C)水平极显著高于对照组奶牛,血糖水平显著低于对照组奶牛。二元Logistic回归分析显示:产后3 d血液TC、HDL-C、K、BHBA、泌乳量与奶牛酮病的发生显著相关。将显著性相关指标利用Logistic回归建模,最终建立了基于血液HDL-C、BHBA、泌乳量的奶牛酮病早期诊断模型,即y=17.95-9.81×HDL-C-2.71×BHBA-0.20×泌乳量,当y<0时存在发生酮病风险,模型早期预测奶牛酮病的准确率82.8%、灵敏度81.3%、特异性84.6%。     

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.     

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.     

Plasma hormone changes in cows during induced or spontaneous calvings and the early post partum period

Plasma progesterone, oestradiol, 13,14-dihydro, 15-keto, prostaglandin F2 alpha (PGFM) and luteinising hormone were measured in daily samples from eight dairy cows for 15 days before and after spontaneous calving, and compared to patterns obtained from 15 cows during and after induction of parturition with dexamethasone +/- prostaglandin. The average (+/- sd) length of pregnancy in the control cows was 285 +/- 3.04 days compared to 264 +/- 7.4 days in the induced group. Only nine cows required prostaglandin on day 11 after dexamethasone, which resulted in all nine calving 1.7 +/- 0.3 days later. Eight of the 15 treated cows retained the fetal membranes; however, the calving to first heat interval for all treated cows was 46.7 +/- 10 days (range 30 to 66 days) and calving to conception 52.4 +/- 17 days (range 30 to 85 days) compared to 42.1 +/- 15 days (range 28 to 64 days) and 76.5 +/- 24 days (range 54 to 115 days) respectively for the eight cows which calved without induction. The pre-partum hormone profiles were similar in all cows. Hormone profiles in the post partum period were also similar in all cows, irrespective of previous history. Baseline values for progesterone and oestradiol were not reached until 4.6 +/- 1.7 days and 2.75 +/- 0.7 days post partum respectively, whereas PGFM values increased for two or three days, reaching baseline levels 8.87 +/- 2.10 days later. Five cows had secondary peaks of PGFM on days 5,6 and 7. Luteinising hormone values increased steadily after calving in all cows.(ABSTRACT TRUNCATED AT 250 WORDS)     

Term placenta shows methylation independent down regulation of Cyp19 gene in animals with retained fetal membranes

Retention of fetal membranes (RFM) is the major post-partum disorder in dairy cattle. Cyp19 gene encodes the aromatase enzyme responsible for catalyzing the rate limiting step in estrogen biosynthesis, an important hormone for placental maturation and expulsion. The present study was aimed for comparative analysis of Cyp19 gene expression and its epigenetic regulation in placental cotyledons of animals with and without RFM. Significantly lower expression of Cyp19 gene was found in placental samples of RFM affected animals in comparison to normal animals. Methylation analysis of 5 CpG dinucleotides of placenta specific Cyp19 gene promoter I.1 and proximal promoter, PII showed hypo-methylation of both PI.1 and PII in term placenta of normal and diseased animals. In conclusion, a mechanism other than promoter methylation is responsible for decreased aromatase expression in placental cotyledons of animals suffering from RFM.     

"益母生化散"对胎衣不下奶牛胎盘激素含量的影响

随机选择1～4胎龄健康黑白花奶牛10头,设为健康对照组(A组);选择1～4胎龄产后健康黑白花奶牛17头,产后立即灌服"益母生化散"250 g,为中药预防组(B组);选择1～4胎龄产后胎衣不下奶牛10头,为胎衣不下组(C组);选择1～4胎龄产后胎衣不下奶牛26头,产犊后12 h灌服"益母生化散"500 g,设为中药治疗组(D组).采用酶联免疫吸附法(ELISA)检测奶牛胎盘组织中孕酮、雌激素、前列腺素和血管紧张素的含量,探讨奶牛胎衣不下的发生机理及中药"益母生化散"的防治效果.结果表明,奶牛胎衣不下的发生与胎盘组织中孕酮、雌激素、前列腺素(PGE2)和血管紧张素Ⅱ含量变化关系密切,胎盘组织中孕酮含量的升高,雌激素、前列腺素和血管紧张素含量的降低是胎衣不下发生的重要机理之一."益母生化散"对奶牛胎衣不下有较好地防治效果,其机制可能与"益母生化散"具有调节胎盘组织的激素分泌等作用有关.     

Metabolic changes in early lactation and impaired reproductive performance in dairy cows

This review addresses the suggestion that the decline in dairy reproductive performance, as increasingly observed these days, may be due to a hampered process of metabolic adaptation in early lactating cows. In our opinion, adaptation to the negative energy balance is a gradual process. Because almost all cows do adapt in the long run, it is not possible to classify animals as adapted or non-adapted. The use of risk factors is more appropriate in this case and is discussed in this review. Among them are the body condition score and its derivatives, feed intake, the calculated negative energy balance, and metabolic parameters like the plasma concentration of insulin or the triacylglycerol content in the liver. Moreover, factors that play a role in the link between declined reproductive performance and the metabolic situation of the cow during the early lactating period are discussed. Among these are insulin, insulin-like growth factors, leptin, neuropeptide Y, non-esterified fatty acids, thyro?d hormones, urea, and ammonia.     

胎衣不下奶牛血浆中矿物质及维生素含量变化

选取胎衣不下组和胎衣正常排出组奶牛各6头,通过终点法对两组奶牛血浆中矿物质指标Ca、P、Fe、Zn的含量检测,同时采用高效液相色谱法测量两组奶牛血浆中维生素指标VA、VD、VE的含量。结果表明,胎衣不下组奶牛血浆中矿物质Ca、Zn、Fe的含量较胎衣排出组正常奶牛低,而血浆中P的含量较正常奶牛的高;胎衣不下组奶牛血浆中维生素VD和VE的含量低,而血浆VA的含量与胎衣正常排出组奶牛无差异性。     

Levels of oestrone sulphate during pregnancy in different breeds of cows and its possible association with retained foetal membranes.

The levels of oestrone sulphate in plasma of pregnant cows was followed from 30 days of pregnancy until parturition. The Swedish Jersey Breed (SJB) showed significantly lower levels of oestrone sulphate between 101 and 200 days of pregnancy than either the Swedish Red and White (SRB) or the Swedish Lowland Breed (SLB). No significant difference was noted between SRB and SLB. On days 141-160 of gestation the oestrone sulphate values were still below the 10 nmol/l level for the SJB while they were above this level for the SRB and the SLB, and the difference was significant. In the SJB, levels above 10 nmol/l were reached on days 161-180 of gestation. In the second part of this study the levels of oestrone sulphate were measured around parturition in SRB cows. At parturition, the levels of oestrone sulphate rose to peak values of 79.9 +/- 5.2 nmol/l and then decreased to 6.6 +/- 0.5 nmol/l on the day after calving. In one cow peak values of 66.0 nmol/l were reached 2 days prior to parturition, and subsequently dropped to 7.0 nmol/l at parturition. This cow had retained foetal membranes. A possible relationship between low oestrone sulphate levels prior to parturition and retained foetal membranes is discussed.