A Case of Successful Pregnancy in a Ewe with Uterus Didelphys

Uterus didelphys is a rare congenital abnormality of the reproductive tract. Although it occurs in various species, there are no published reports describing pregnancy outcome in association with this abnormality. Herein we describe a case of successful unilateral singleton pregnancy in a ewe incidentally found to have uterus didelphys during the course of a biomedical research study. The pregnancy was established using assisted reproductive techniques and interrupted in late gestation, at which point the abnormality was identified. Serial ultrasound assessment of foetal biometry revealed a normal foetal growth trajectory. Despite a 45% reduction in placentome number, total placentome weight was near normal secondary to compensatory placentome growth and development. To our knowledge, this is the first detailed report of normal foetal growth in an animal with uterus didelphys and illustrates the ability of the ovine placenta to adapt to a reduced number of placentomes and maintain foetal nutrient supply.     

Assessment of the accuracy of estimation of gestational age in cattle from placentome size using inverse regression

AIMS: To assess, in dairy cows between 60–160 days of gestation, whether the agreement between actual gestational age and that predicted by placentome size could be improved by using individual placentome measurements rather than a single average, and to identify how increasing the number of placentomes measured improved the prediction.

METHODS: Data were obtained from 58 cows examined using transrectal ultrasonography every 10 days between 60–130 days, and every 15 days between 130–160 days of gestation. For each cow, four to six placentomes in each of the pregnant and non-pregnant horns were examined from the region of the uterus near to the cervix. A mixed-effects model, which included cow as a random effect, and a simple linear regression which ignored within-cow correlation, were fitted to the data. Inverse regression was used to compare the 95% prediction bands obtained for estimating gestational age using the means of three, five or eight placentome measurements.

RESULTS: The fit of the mixed effects model was better than a simple linear regression (p<0.001) but the fitted lines from the two models were very similar. Using the simple regression model, for a mean placentome length of 15?mm estimated gestational age was 100 days, with 95% CI of 68–131 days for measurement of three placentomes, and 80–120 days for eight placentomes.

CONCLUSIONS: The agreement between placentome size and gestational age was improved by increasing the number of placentomes measured. Direct comparison of these CI with those for other published fetal measures was not possible as similar prediction bands have not previously been calculated for fetal measures; however one study reported a residual SD which had been calculated using size measurements as the predictor variable and age as the response variable. Using these data 95% CI were calculated to be ±9 days for crown-rump length and ±25 days for uterine diameter. These are likely to be an underestimate of the true CI and do not take account of the increase in variance of the difference between predicted and actual gestational age as gestational age increases.

CLINICAL RELEVANCE: Placentome measurement may be useful for estimating fetal age in late gestation. Further research is required to better establish the agreement between gestational age and the fetal measurements which are currently used to estimate fetal age.     

Effect of prostaglandin F2 alpha-induced luteolysis on in vivo and in vitro progesterone production by individual placentomes of cows

This study investigated placental progesterone production by bovine placentomes. Catheters were placed in the femoral artery (FA) and in the caruncular artery (CA), caruncular vein (CV) and lymphatic vessel of a prominent placentome of 13 cows at 200 d of gestation. Four of the 13 cows were given prostaglandin F2 alpha (PGF2 alpha) after surgery, and blood and lymph were collected for progesterone determination. After 24 h, progesterone was higher (P less than .01) in FA and CA plasma from control cows that FA and CA plasma from PGF2 alpha-treated cows (5.11 +/- .29 and 5.17 +/- .64 vs 1.41 +/- .08 and 1.15 +/- .08 ng/ml, respectively), but CV concentrations were similar (3.38 +/- .30 vs 2.56 +/- .24, respectively). There was a net uptake of progesterone by placentomes from control cows (P less than .01) but a net secretion in PGF2 alpha-treated cows (P less than .05). Lymph contained low progesterone concentrations regardless of treatment. Cows were slaughtered at 240 d of gestation. Placentomes were removed and perfused with pregnenolone through the maternal and fetal arteries. Fetal venous effluent contained more progesterone than maternal venous effluent (P less than .001) in both groups, and fetal venous effluent of placentomes from PGF2 alpha-treated cows contained more progesterone than that from control cows (P less than .05). Maternal and fetal components of other placentomes were cultured alone or in co-culture along with pregnenolone and (or) epostane. Fetal tissue produced more progesterone (P less than .001) than maternal tissue when each was cultured alone, but fetal tissue production declined when co-cultured with maternal tissue.(ABSTRACT TRUNCATED AT 250 WORDS)     

Expression of vascular endothelial growth factor and endothelial nitric oxide synthase is increased in the placenta of sheep at high altitude in the Andes

Fetal weight and the placenta of sheep at high altitude (HA) are affected by hypoxia. Placental changes (an increase in placental size and vascularization) are greater in ewes from populations that have lived for several generations at HA than in those exposed during just 1 gestation. This study investigated placental expression of vascular endothelial growth factor (VEGF) and endothelial nitric oxide synthase (eNOS), 2 molecules involved in placental angiogenesis that could be upregulated by hypoxia. Two groups of ewes were maintained at HA (3589 m) during pregnancy: HA-native ewes (group HH) and ewes native to lowlands but moved to HA immediately after the diagnosis of pregnancy (group LH). A control group (LL) was kept at sea level. Near term, placentomes were removed, weighed, and processed for immunohistochemical detection of VEGF and eNOS, as well as for vascular area measurement. Placental weight was significantly higher in the HH group than in the LH and LL groups; between the latter 2 groups there was no significant difference. The placental area occupied by vasculature was significantly greater in both the HA groups than in the LH group; the number of placentomes was greatest in the LL group. The density of VEGF and eNOS in the placentome tissue was significantly greater in both HA groups than in the LL group. Although the density of VEGF was significantly lower in the HH group than in the LH group, no differences were observed in eNOS density between the HH and LH animals. These results demonstrate that chronic hypoxia upregulates the expression of placental VEGF and eNOS, suggesting an important role of these molecules in the placental response to HA hypoxia. In addition, an attenuated response to hypoxia in VEGF synthesis may be part of the long-term process of adaptation to HA.     

妊娠后期限饲蒙古绵羊对其胎盘生长发育及胎盘块类型的影响

本试验旨在研究妊娠后期限饲蒙古绵羊对其胎盘生长发育及胎盘块类型分布的影响。选择健康蒙古绵羊42只(经同期发情且受孕),于妊娠90 d选择6只母羊进行屠宰,其余按体重随机分配到3个组:0.175 MJ/(kgW0.75.d)(RG1:n=14)、0.33 MJ/(kgW0.75.d)(RG2:n=12)和自由采食组0.67 MJ/(kgW0.75.d)(CG:n=10),进行不同能量水平饲养至妊娠140 d,再选择6只母羊进行屠宰。结果表明:妊娠后期限饲母羊严重影响了RG1组(P<0.01)、RG2组(P<0.05)的胎盘重及胎儿重(P<0.01);RG1组A类型胎盘块重量(P<0.01)、数量(P<0.01)与RG2组A类型胎盘块数量(P<0.05)显著低于CG组;RG1组B类型胎盘块重量(P<0.05)、数量(P<0.01)与RG2组B类型胎盘块数量(P<0.05)显著高于CG组;随着母体营养水平的降低,C、D类型胎盘块出现的几率增加,且C、D类型胎盘块血管数量显著高于A、B类型胎盘块血管数量(P<0.05)。这说明妊娠后期限饲蒙古绵羊影响了胎盘的生长发育,使得胎盘块数量、重量、类型和血管数量等发生了明显改变。     

Relationships between uterine and fetal traits in rabbits selected on uterine capacity

The aim of this study was to investigate whether uterine capacity (UC) in rabbits was related to uterine horn length and weight and whether these uterine traits and vascular supply were related to fetal development and survival. Data from 48 unilaterally ovariectomized (ULO) does of the High and 52 ULO does of the Low UC lines of a divergent selection experiment on UC were used. Does were slaughtered on d 25 of fifth gestation. The High line showed higher ovarian weight (0.08 g, P < 0.05) linked to a higher ovulation rate (1 ovum, P < 0.05) and greater length of the empty uterine horn. There were no differences between lines in the remaining doe traits. The number of implanted embryos and live fetuses, fetal survival, and uterine weight and length were positively associated and explained most of the observed variation. Average weights of the live fetuses and their fetal and maternal placentae were not related to uterine weight and length. The linear regression coefficient of full uterine horn length on the number of live fetuses was 2.43 +/- 0.21. The weight of the full uterine horn showed a small quadratic relationship (P < 0.05) with the number of live fetuses. Full uterine horn length, after adjusting for the number of embryos, was negatively associated (P < 0.001) with the number of dead fetuses. The linear regression coefficient of average fetal placental weight of the live fetuses on number of implanted embryos was higher (P < 0.10) in the Low line (-0.23 +/- 0.04 vs. -0.12 +/- 0.04). The linear regression coefficient of average weight of the live fetuses on the average weight of their fetal placentae was higher (P < 0.10) in the High line (2.56 +/- 0.47 vs. 1.27 +/- 0.57). The High line was more efficient, most likely because an increase in intrauterine crowding has a lesser effect on the development of fetal placentae and fetuses. The fetal position within the uterus did not affect the proportion of dead embryos. Fetuses with placentae receiving a single blood vessel had a higher probability of death (P < 0.001) and the lowest weight. There was no difference between lines for individual weight of the live fetuses, but the High line showed higher individual weights of fetal (P < 0.01) and maternal placentae (P < 0.10). Live fetuses in the midportion of the uterus were lighter in weight (P < 0.05) than in the oviductal and cervical regions (20.3 vs. 21.6 and 21.7g). Increasing uterine capacity increases uterine length and decreases weights of fetus and fetal placenta in rabbits.     

Conceptus competition for uterine space: different strategies exhibited by the Meishan and Yorkshire pig

Our laboratory has demonstrated that Yorkshire placentae increase in size and surface area during the final third of gestation. In contrast, Meishan placental size remains constant during late gestation, but the density of blood vessels at the placental-endometrial interface increases markedly. Preliminary observations from our laboratory suggest that if one of two adjacent Meishan fetuses dies, the placenta of the remaining Meishan conceptus fails to increase its length of implantation or its placental weight or surface area. In contrast, if one of two adjacent Yorkshire fetuses dies, the adjacent conceptus accelerates its placental growth. The objective of this experiment was to document that Yorkshire, but not Meishan, conceptuses accelerate placental growth when adjacent fetuses are experimentally destroyed on d 40 of gestation. Straightbred Meishan (n = 5) and Yorkshire (n = 5) females were laparotomized and one uterine horn was randomly assigned to receive fetal crushing (treated horn); the other uterine horn served as a within-animal control. In the treated horn, every other fetus was then crushed through the uterine wall and the animals were allowed to recover. On d 111, animals were killed, uteri were recovered, and fetal weight, crown-rump length (CRL), placental weight, implantation site length, and placental surface area were recorded. Although there were no statistically significant differences in fetal weight or CRL observed between treated or control horns of females of either breed, there was a tendency for the fetuses in the treated uterine horn to be longer and heavier in both breeds. There were no differences in placental weight, placental surface area, or implantation site length between conceptuses in Meishan treated and control horns, which averaged 173.8+/-6.4 g, 1,162.7+/-35.9 cm2, and 19.0+/-0.4 cm, respectively. In contrast, placental weight, placental surface area, and implantation site length were increased (P < 0.05) in Yorkshire treated horns compared to Yorkshire control horns (306.1+/-26.0 g, 1,835+/-93.9 cm2, and 33.4+/-1.5 cm vs 253.7+/-13.4 g, 1,474.3+/-50.4 cm2, and 27.2+/-0.8 cm; respectively). These data confirm that Yorkshire conceptuses, but not Meishan conceptuses, accelerate placental growth when adjacent littermates perish as late as d 40 of gestation. These data indicate that differences exist in the strategies employed by Meishan and Yorkshire conceptuses in the competition for nutrients during gestation.     

Ultrasound during mid‐gestation: Agreement with physical foetal and placental measurements and use in predicting gestational age in sheep

To determine the effects of poor maternal nutrition and litter size on foetal growth during mid‐gestation, pregnant ewes (n = 82) were fed 100%, 60% or 140% of NRC TDN beginning at day 30.2 ± 0.2 of gestation. Transabdominal ultrasound was performed weekly between day 46.0 ± 0.4 and 86.0 ± 0.7 to monitor foetal heart width (HW), umbilical diameter (UMB), rib width (RW) and placentome outer (OD) and inner diameter (ID). Data were analysed with repeated‐measures using the mixed procedure for effects of maternal diet, litter size and gestation, and equations predictive of gestational age were generated using the regression procedure. To determine the agreement of ultrasound measurement and actual size, ewes (n = 20–21) were euthanized at day 45 or 90 to obtain corresponding postmortem measurements for Bland–Altman analysis. The HW, UMB and placentome OD and ID increased with gestation (p < .0001) but were unaffected by maternal diet or litter size (p ≥ .12). Ultrasound underestimated postmortem measurements of HW (14.8%), UMB (7.3%), placentome OD (4.5%) and ID (37.3%) at day 90 of gestation. Ultrasound underestimated RW at day 45 (7.7%) but overestimated RW (23.8%) at day 90, indicating inconsistent bias when reporting RW by ultrasound. Combining the HW, UMB, RW and placentome OD generated the strongest equation predictive of gestational age (R² = .91). These findings indicate that during mid‐gestation, maternal diet or litter size did not affect HW, UMB or placentome diameters and these factors can be used to estimate gestational age.     

Acoustic radiation force impulse (ARFI) elastography to asses maternal and foetal structures in pregnant ewes

The aim of this study was to evaluate the stiffness of foetal lungs, liver, kidneys and placentomes by acoustic radiation force impulse (ARFI) elastography in ovine and to correlate obtained findings with foetal development. Twenty‐four ewes were included, and tissue stiffness of foetal lung, liver, kidney and placentome was measured beginning with 10th gestational week by qualitative–quantitative ARFI elastography. A total of 33 healthy lambs were born. Qualitative elastographic analysis permitted to classify maternal and foetal tissues elasticity in decreasing order as follows: placentome, kidney, liver and lung. Regarding quantitative ARFI elastography data, shear wave velocity (SWV) of foetal lung and liver varied. The lung SWV decreased gradually from the 16th to the 21st gestational week (R² = 0.80; p < 0.001), while liver SWV increased gradually from the 14th to 21st gestational week (R² = 0.80; p < 0.001). In contrast, the kidneys and placentomes SWV's remained constant through gestation (p = 0.076; and 0.34). ARFI elastography was shown feasible for evaluation of maternal and foetal tissues stiffness in the ovine model. It can be used to demonstrate pulmonary and hepatic stiffness modifications during foetal development.     

A stereological method for estimating the feto-maternal exchange surface area in the bovine placentome at 135 days gestation

The feto-maternal relationships within the bovine placentome at 135 days gestation were investigated using unbiased stereology. This provides an estimate of the feto-maternal interface without physically separating these. The functional surface area, the volume of the fetal and maternal components, volumetric tissue shrinkage due to processing, and the feto-maternal surface amplification factor were estimated and quantified in standard units. In this healthy bovine pregnancy there were 105 placentomes with a total volume of 900 ml.The volumetric tissue shrinkage was 24% and the maternal: fetal tissue ratio overall was 0.92. The total feto-maternal exchange surface area was 18.5 m² with a surface amplification factor of 108.1. Analysis of the coefficient of error from counting different numbers of points within the reference area suggested that 50% and 75% fewer points could be counted to achieve the result obtained by counting approximately 6000 points used in this study.     

Kinetics of infection and effects on the placenta of Chlamydophila abortus in experimentally infected pregnant ewes

A Chlamydophila abortus-induced abortion model was carried out on the basis of the experimental infection of ewes at day 75 of gestation. The infection induced abortions and the birth of weak lambs during the last 3 weeks of pregnancy. To study the kinetics of the infection in the placenta and in other organs, infected ewes were killed at 105, 120, and 130 days of gestation and also several days after abortion or parturition. Infected ewes developed a systemic infection that caused a mild and transient pneumonia and focal hepatitis. Pathologic changes were observed in placentas at 120 day of gestation, although the lesions varied between animals and even between placentomes of the same placenta. The first placental area infected was the maternal stroma and epithelium next to the intercaruncular areas, where neutrophilic response seemed to control the infection. A substantial degree of multiplication of C. abortus was then observed in the trophoblast cells of the placentome, periplacentomal choriallantoic membranes, and hilius, with an inflammatory exudate composed mainly of neutrophils, some macrophages, and very scarce lymphocytes. After abortion, the lesions affected the intercotyledonary areas of the aborted placentas, whereas in the uterus significant lymphocyte infiltration was observed, together with a rapid decrease of the C. abortus antigen in the degenerated caruncular tissues.     

应用B-超对小尾寒羊早期胚胎和胎盘发育的研究

应用B-超监测10只健康的妊娠小尾寒羊,以研究早期胚胎和胎盘的发育过程。结果表明,早期胚体发育在妊娠第6周完成;妊娠30～46d,羊膜囊的短径与胎龄之间存在线性关系(R=0.9528,P<0.01),回归方程为Y=0.565065X 18.751970(Y:妊娠天数,X:羊膜囊宽)。胎盘发育期集中在妊娠前11周,在此期间胎盘突直径也可用于预测胎龄,二者之间的方程式为:Y=4.961429X-16.655714(Y:胎盘突直径,X:妊娠周),相关系数R=0.9882(P<0.01)。     

Distribution of viral antigen in uterus, placenta and foetus of cattle persistently infected with bovine virus diarrhoea virus

The tissue distribution and cellular localisation of bovine virus diarrhoea virus (BVDV) was investigated in the uterus, placentomes, intercotyledonary foetal membranes and foetal organs of three persistently infected (PI) pregnant heifers. The uterus and ovaries of a non-pregnant PI heifer were also included in the study. Cryostat sections were examined using immunohistochemical techniques and monoclonal antibodies against BVDV. A double immunofluorescence technique was used to identify BVDV positive cells that also showed staining for either the leukocyte common antigen CD45 or the cytoskeletal filament vimentin. BVDV antigen was detected in all the organs examined, and was present in both epithelial and non-epithelial cells. In all organs many of the virus-positive cells also showed reactivity for vimentin. In the foetal liver and spleen a small, scattered population of virus-positive cells showed reactivity for CD45. A few cells showed reactivity both for BVDV antigen and for CD45 in the placentomes and intercotyledonary foetal membranes. In contrast to earlier reports, only scattered cells in the foetal part of the placentomes, the cotyledons, showed reactivity for BVDV antigen. However, in the chorion of the intercotyledonary foetal membranes, a larger proportion of the trophoblast cells showed reactivity for BVDV, especially the binuclear trophoblast cells. In the uterus, pregnancy appeared to favour virus replication, as the section from the pregnant heifers showed much stronger staining and a higher proportion of viral antigen-positive cells than sections from the non-pregnant PI heifer.     

Placentomal differentiation may compensate for maternal nutrient restriction in ewes adapted to harsh range conditions

Maternal nutrient restriction from early to midgestation can lead to fetal growth retardation, with long-term impacts on offspring growth, physiology, and metabolism. We hypothesized that ewes from flocks managed under markedly different environmental conditions and levels of nutrition might differ in their ability to protect their own fetus from a bout of maternal nutrient restriction. We utilized multiparous ewes of similar breeding, age, and parity from 2 flocks managed as 1) ewes adapted to a nomadic existence and year-long, limited nutrition near Baggs, WY (Baggs ewes), and 2) University of Wyoming ewes with a sedentary lifestyle and continuous provision of more than adequate nutrition (UW ewes). Groups of Baggs ewes and UW ewes were fed 50 (nutrient restricted) or 100% (control fed) of National Research Council recommendations from d 28 to 78 of gestation, then necropsied, and fetal and placental data were obtained. Although there was a marked decrease (P < 0.05) in fetal weight and blood glucose concentrations in nutrient-restricted vs. control fed UW ewes, there was no difference in these fetal measurements between nutrient-restricted and control-fed Baggs ewes. Nutrient-restricted and control-fed UW ewes exhibited predominantly type A placentomes on d 78, but there were fewer (P c0.05) type A and greater (P < 0.05) numbers of type B, C, and D placentomes in nutrient-restricted than control-fed Baggs ewes. Placental efficiency (fetal weight/placentomal weight) was reduced (P = 0.04) in d 78 nutrient-restricted UW ewes when compared with control-fed UW ewes. In contrast, nutrient-restricted and control-fed Baggs ewes exhibited similar placental efficiencies on d 78. This is the first report of different placental responses to a nutritional challenge during pregnancy when ewes were selected under different management systems. These data are consistent with the concept that Baggs ewes or their conceptuses, which were adapted to both harsh environments and limited nutrition, initiated conversion of type A placentomes to other placentomal types when subjected to an early to mid-gestational nutrient restriction, whereas this conversion failed to occur in UW ewes. This early placentomal conversion in the Baggs ewes may function to maintain normal nutrient delivery to their developing fetuses during maternal nutrient restriction.     

Amyloid A in equine colostrum and early milk

The objective of this study was to investigate the protein, amyloid A3 (AA3), in equine colostrum and early milk. We hypothesized that AA3 was consistently present in equine colostrum and early milk, that no correlation existed between serum and colostrum concentrations of this protein in individual mares at parturition and that colostrum/milk concentrations of this mammary protein may be affected by age, breed, length of gestation and/or induction of parturition. Thirty-eight peripartum mares and seven non-pregnant, non-lactating mares were included in the study. Mean serum concentrations of this protein in the pregnant and non-pregnant mares were consistent with previous reports. Amyloid A3 was found in all colostrum and early milk samples at consistently higher concentrations than in peripartum maternal serum. There was no correlation between serum AA and colostrum AA3 concentrations at parturition. Age and breed effects were not significant. Increased gestation length and induction of parturition were associated with decreased colostrum and milk AA3 concentrations. We conclude that AA3 is consistently present in equine colostrum and early milk. The production of this protein in the mammary gland is likely to be under different stimulus to the production of serum AA, and may have protective effects in the neonatal intestine.     

Ovulation rate and embryo survival during gestation and the effects of ovariectomy on maternal nurturing ability

Correlated responses to selection for increased litter size were studied in mice. The selected line (L+) was compared with an unselected control line (K') in two experiments. The first experiment provided a profile of correlated changes in female reproductive traits at d 0, 6, 14 and 18 of gestation. Experiment two examined the effects of ovariectomy on d 18 of gestation, sham surgery and no surgery on litter size and maternal performance. Females of the L+ line had increased (P less than .001) body weight, ovulation rate and uterine length at d 0 of gestation compared with K' females, but uterine weight and ovarian weight did not differ. Positive correlated responses (P less than .001) in uterine weight and length at d 6 and 14 of gestation were associated with a larger number of viable fetuses. Space per fetus was reduced (P less than .001) in the uterus of L+ females, but a lower fetal mortality was still maintained in L+ throughout gestation. Prenatal survival was about 10% higher (P less than .06) in L+, the major difference (P less than .01) occurring before implantation. A second experiment was conducted to determine the effect of ovariectomy on d 18 on litter size and maternal performance. In experiment two, no significant line X treatment interactions were found for maternal performance, indicating that both lines responded similarly to ovariectomy. Line L+ showed a positive correlated response in maternal performance. Ovariectomized females had a reduced (P less than .05) number born alive compared with sham-operated females, but the nonsurgically treated females were intermediate.(ABSTRACT TRUNCATED AT 250 WORDS)     

Serum cortisol and progestin concentrations in pregnant and non-pregnant Asian elephants (Elephas maximus)

Blood samples were collected during the estrous cycle (n=3), throughout gestation (n=3), and during the periparturient period (n=11) to assess serum concentrations of cortisol in pregnant and non-pregnant Asian elephants whose reproductive status was being monitored by serum progestin determination. While serum cortisol concentrations remained constant throughout gestation, progestin concentrations decreased significantly (p<0.05) in the second half of pregnancy, declining to undetectable levels by 3 days before calving. During the non-luteal phase of the estrous cycle serum progestins varied from undetectable levels to 100pg/ml (53+/-10.7pg/ml) then increased steadily during the luteal phase (322+/-207.5pg/ml). There were no significant differences between serum cortisol concentrations during the luteal and those of the non-luteal phase (p>0.05). The mean cortisol concentration during the estrous cycle was about twice that during pregnancy (p>0.05). No substantial changes in maternal cortisol were found during the course of pregnancy or the periparturient period.