Transfer of Beef Embryos in Dairy Cows: Influence of Recipient and Embryo Quality on Pregnancy Rate and Calving Performance

Contents
On three experimental farms 205 deep frozen Blonde d'Aquitaine embryos were transferred into Holstein dairy cows 7 days after a spontaneous heat. The aim of the study was firstly to examine the influence of recipient and embryo quality on pregnancy rate and secondly to study the calving performance of dairy cows carrying beef embryos. Recipients were divided into three groups based on the quality of the corpus luteum (unsuitable, moderately suitable and suitable) established by rectal palpation. Embryos classified as fair, good and excellent, were randomly distributed over the recipients, including those classified as being unsuitable. The overall pregnancy rate was 36% (standard deviation (SD) = 3.3), dystocia occurred in 48% of births (SD = 5.9), gestation was 297 days (SD = 1.2) and birth weight was 52 kg (SD = 2.0). No statistical differences were found in pregnancy rates and calving performance between the groups of recipients or between the classes of embryos. It is concluded that there is no justification to reject recipients on the basis of the quality of the corpus luteum. Furthermore there is no reason to reject fair embryos for transfer. Beef embryos in dairy cows resulted in much dystocia because of high birth weight, and in long gestation periods.     

Therapeutic Approaches to Pregnancy Loss of Non‐infectious Cause During the Late Embryonic/Early Foetal Period in Dairy Cattle. A Review

The problem of pregnancy loss during the late embryonic/early foetal period due to non‐infectious causes in dairy cattle is on the rise, especially in high milk producing cows. This review discusses recent therapeutic approaches designed to reduce the incidence of early foetal loss. These strategies are based on the fact that low plasma concentrations of progesterone are commonly related to foetal loss. Progesterone supplementation during the late embryonic/early foetal period has proven useful in curtailing losses, mainly in single pregnancies, whereas a more positive effect of treatment with GnRH than progesterone has been found in twin pregnancies. Therapeutics is not necessary in cows with an additional corpus luteum. In practical terms, suggestions include recording the number of embryos and corpora lutea on first pregnancy diagnosis and checking for normal pregnancy progression on day 60 post‐insemination.     

Factors Affecting Plasma Pregnancy-associated Glycoprotein 1 Concentrations Throughout Gestation in High-producing Dairy Cows

This study was designed to establish the factors, if any, which could affect plasma pregnancy-associated glycoprotein-1 (PAG-1) expression in a study population of 87 pregnant, high-producing dairy cows. The factors examined were: semen providing breed (Holstein-Friesian vs Limousin), outcome of gestation (male vs female newborn, and singleton vs twin pregnancies), lactation number, milk production at pregnancy diagnosis, plasma progesterone concentration, season of gestation (warm period, March–November vs cool period, December–February), and day of gestation (40, 90, 120, 150, 180 and 210). Pregnancy was diagnosed by transrectal ultrasound on day 40 post-insemination and by palpation per rectum on days 90, 120, 150, 180 and 210. Blood samples were collected from each animal immediately before each pregnancy diagnosis. The relative contributions of the different factors on PAG-1 concentrations were evaluated by GLM repeated measures analysis of variance. No significant effects of the herd, foetal sex, milk production, lactation number and plasma progesterone concentrations were observed. In contrast, twin pregnancy, the use of Limousin semen and conception during the cool period were correlated with significantly increased plasma PAG-1 concentrations throughout gestation. Our data indicate that both cow well-being during early placental development, determined in our conditions by reduced heat stress when conception occurred in the cool season, and crossbreed pregnancies lead to improved PAG-1 production throughout the gestation period.     

Timing of Early Foetal Loss for Single and Twin Pregnancies in Dairy Cattle

The incidence of early foetal loss is increasing under intensive management systems for dairy cattle. The aims of the present study were to determine whether there is any peak period of pregnancy loss during the early foetal period and to evaluate possible differences between single and twin pregnancies. The study population consisted of 1442 pregnant cattle from a single herd. Pregnancy was diagnosed by transrectal ultrasonography between 36 and 42 days after insemination, and then weekly until day 90 of gestation or until pregnancy loss. A total of 1310 cows (90.8%) bore single embryos and 132 (9.2%) carried twins. Pregnancy loss was registered in 139 (9.6%) cows before day 90 of pregnancy: 101 (7.7%) in single and 38 (28.8%) in twin pregnancies. The average time of pregnancy loss for all animals was 58.4 ± 12.6 days and ranged from 45 to 90 days. Seventy‐five per cent of the pregnancy losses were registered between 45 and 60 days of gestation. The average time of pregnancy loss for cows with singletons was 52.1 ± 4.1 days and ranged from 45 to 61 days and that for those with twins was 75.1 ± 12.4 days and ranged from 46 to 90 days. Seventy‐five per cent of the twin pregnancy losses were registered between 68 and 90 days of gestation. Our data show that the foetal loss in singleton pregnancies occurs earlier than in twin pregnancies. Assessment of normal development of gestation on days 60 and 90 after insemination is suggested.     

Manual Rupture Versus Transvaginal Ultrasound-guided Aspiration of Allanto-amniotic Fluid in Multiple Pregnancies: A Clinical Approach to Embryo Reduction in Dairy Cattle

To avoid the problems associated with twinning in dairy cattle, one of the embryos may be eliminated. This study compares the effect on pregnancy maintenance of two embryo reduction techniques, manual rupture (MR) and transvaginal ultrasound-guided aspiration (TUGA) of allanto-amniotic fluid, in Holstein-Friesian cows with multiple pregnancies. In the first experiment, 61 lactating cows bearing unilateral twins (n=27), bilateral twins (n=30) or triplets/quadruplets (n=4) were subjected to MR (n=45) or TUGA using a 17-G neddle (n=16) on day 28-34 of gestation. In 21 and 10 cows undergoing MR and TUGA embryo reduction, respectively, pregnancy loss occurred before day 90 (46.7 vs. 62.5%, P= 0.28). Through binary logistic regression, the type of pregnancy was identified as the only variable significantly affecting pregnancy maintenance (P=0.03). Based on the odds ratio, the risk of pregnancy loss was 4.1 times higher for unilateral twins than for bilateral twins (70.4 vs. 36.7%, respectively, P=0.01). No effect was detected on pregnancy maintenance of the technique used (P=0.17) or of the interaction technique by type of pregnancy (P=0.22). In the second experiment, a 22-G needle was used to perform TUGA on 22 lactating cows. The pregnancy loss rates were 44.4% (4/9), 18.2% (2/11) and 50% (1/2) for cows bearing unilateral twins, bilateral twins and triplets, respectively. The total pregnancy loss rate following TUGA using the 22-G needle tended to be lower than that using the 17-G needle (31.8 vs. 62.5%; P=0.06). Our results suggest that TUGA using a 22-G needle could be the method of choice to perform embryo reduction in cows carrying multiple pregnancies.     

Plasma oestrone and oestradiol concentrations throughout gestation in cattle: relationship to stage of gestation and fetal number

The objectives of the study were to characterise the peripheral plasma oestrone (E1) and oestradiol-17beta (E2) concentrations throughout gestation in the cow and to correlate this with the stage of gestation and fetal number. Cows (n = 10) were equally divided into two groups after non-surgical embryo transfer of in-vitro matured and in-vitro fertilised (IVM - IVF) embryos; Group 1 received a single embryo, Group 2 received twin embryos. Blood was collected about every third day from day 0 (day 0 was defined as first day of standing oestrus), then daily for the last 10 days of gestation and sampling was stopped one day post partum. Plasma E1 concentration exceeded that of E2 throughout gestation in both groups of cows. The time trend concentrations of plasma E1 were significantly affected by the stage of gestation (P < 0.01) and fetal number (P < 0.01) in the last two trimesters of gestation. The time trend concentrations of plasma E2 were significantly affected by the stage of gestation (P<0.01) but not foetal number (P = 0.09). In both groups there was marked preparturient increase in E1 and E2 concentrations. Plasma E2 profile between days 10 prepartum to parturition paralleled E1 in cows carrying a single foetus but was disparate during the same period in the twin-bearing cows.To conclude, our results indicate that although plasma E1 concentration was greater than E2 throughout gestation, both were related to the stage of gestation and that fetal number was correlated with circulating E1 levels in the last two trimesters of gestation.     

Morphometric study of the porcine placental vascularization

The early development in mammals is characterized by the contribution of nutrients from the maternal tissues through the placenta, which is in apposition with foetal membranes and the endometrium, allowing the physiological interchange between the embryos/foetuses and the mother. The aim of this work was to study the number of placental blood vessels and their vascular area through morphometric analyses and the haemotrophic diffusion distance in porcine placental tissues from early gestations, intermediates gestations, advanced gestations and term gestations. For those purposes, morphometric measurements, blood vessel quantification, high‐resolution light microscopy and transmission electron microscopy were performed. The implementation of the high‐resolution light microscopy allowed studying the placental vascular and tissue histoarchitecture with higher definition and resolution than using a conventional light microscopy. We highlight the close location of the subepithelial capillaries to the maternal/foetal interface as pregnancy progresses. We found statistically significant evidence to state that the area of blood vessels is dependent on the gestation period. In advanced gestations, the presence of numerous small blood vessels and its near location to foetal/maternal interface agree with the great remodelling reported in our previous studies. In conclusion, in gilts, given the type of non‐invasive epithelial placentation, the new blood vessels generation and of haemotrophic diffusion distance reduction, determined in this report, assure the maternal/foetal haemotrophic exchange efficiency during gestation.     

Relationship of ovarian contents to plasma progesterone concentration in the swamp buffalo (Bubalus bubalis)

Using rectal palpation and laparoscopy, the relationship of ovarian contents to plasma progesterone concentration during the oestrous cycle, early pregnancy and post partum periods in the swamp buffalo (Bubalus bubalis) was studied. During the oestrous cycle, four stages in the lifespan of the corpus luteum were seen laparoscopically. The mean (+/- sd) concentrations of progesterone in plasma in cows with and without a corpus luteum on their ovaries were 1.49 +/- 0.78 ng/ml (n = 31) and 0.14 +/- 0.09 ng/ml (n = 14), respectively. Plasma progesterone levels reflected age-dependent changes occurring in the cyclic corpus luteum. The accuracy of diagnosing ovarian contents was 82 and 91 per cent for rectal palpation and plasma progesterone levels respectively. Approximately 29 per cent follicles (larger than 10 mm) were incorrectly diagnosed as corpora lutea by rectal palpation.     

Reciprocal embryo transfer between repeat breeder and virgin heifers--an experimental model

The aim of the study was to evaluate the importance of mother-concaptus relationships for the elevated embryonic loss in repeat breeder heifers. Embryos were collected by non-surgical technique, classified and transferred surgically or non-surgically to synchronized, inseminated recipients. The embryos were transferred to the uterinehorn contralateral to the corpus luteum. The embryos were transferred from repeat breeder heifers (RBH) to virgin heifers (VH) or from VH to RBH. After slaughter 4 weeks after transfer there was no difference in emhryonic survival between heifer categories following transfer or insemination. In some animals degenerated foetal membranes were found in the nonpregnant horn. The study indicates embryonic morphology rather than the category of donor or recipient as influencing the embryonic survival rate.Key words: embryo, embryonic death, embryo transfer, repeat breeder     

Skim milk progesterone in pregnant cows from insemination throughout lactation

The skim milk progesterone profile was assessed by radioimmunoassay, without extraction, from the day of insemination (day 0) until the cows were dried off on day 225 of gestation. A total of 418 samples were collected from 154 pregnant Holstein cows. The daily variation in skim milk progesterone was recorded from day 1 until day 45 of pregnancy to detect the commencement of progesterone secretion from the corpus luteum after insemination. Subsequent determinations were made every 2 weeks from day 46 until lactation ceased. On the day of artificial insemination and for the first 2 days after insemination, all the cows had a basal progesterone concentration <0.1 ng/ml. A rise in progesterone (0.2±0.1 ng/ml) was first detected on the third day after insemination. The progesterone values then increased significantly (p<0.001) until day 15.The values then remained nearly constant (2.5–3.5 ng/ml) until day 106 of pregnancy, when they began to decline. Between days 120 and 180 of gestation, progesterone was significantly decreased (2.2–2.9 ng/ml) before it rose again to the previous plateau (3.5–3.9 ng/ml) around day 180. The progesterone concentration then remained at the higher level until the animals were dried off.Abbreviations AI artificial insemination - RIA radioimmunoassay     

Experimental Infection of Cows with Bovine Viral Diarrhoea Virus in Early Pregnancy – Findings in Serum and Foetal Fluids

Nineteen pregnant cows were experimentally infected with bovine viral diarrhoea virus (BVDV) between day 74 and 81 of pregnancy. All cows became infected and developed serum antibodies. Sixteen of the cows delivered persistently infected (PI) offspring, whereas the remaining three gave birth to calves with detectable serum antibodies and free from BVDV. The 16 cows with PI foetuses developed higher levels of antibodies in serum during pregnancy than did their three peers carrying non‐PI calves. Multivariate analysis showed that the antibody levels in these two groups of cows were significantly different from day 135 of pregnancy. Foetal fluid was successfully collected from 18 of the 19 infected cows and from five uninfected control cows between 10 and 24 days before delivery by use of a percutaneous, blind puncture technique. No negative effects were observed in the cows or their offspring. BVDV was isolated and detected with an immunoperoxidase test in foetal fluid from 13 of the 16 cows carrying PI foetuses, and from 15 of the cows when a quantitative fluorescent polymerase chain reaction (PCR) technique was used. The negative sample in the PCR assay was positive for BVDV antibodies. The number of viral copies per microlitre in foetal fluids varied between 103 and 1080 in the positive samples. All samples taken from the cows carrying non‐PI foetuses were negative for BVDV in both assays. In this experiment, examination of either serum or foetal fluids could identify the cows carrying a PI foetus. Examination of serum for BVDV antibodies was a reliable indicator of a PI foetus if the serum was collected during the last 2 months of pregnancy. For examination of foetal fluids, both viral and serological analyses should be performed. For viral analysis, PCR should be the test of choice. High levels of BVDV antibodies in conjunction with a negative result in the PCR may be indicative of a false‐negative virus result. Further experience with the method of collection of foetal fluids is necessary for evaluation of its safety. Investigation of pregnant cows in order to discover a PI offspring before it is born could be a useful tool in control and eradication of BVDV.     

Experimental infection of cows with bovine viral diarrhoea virus in early pregnancy - findings in serum and foetal fluids

Nineteen pregnant cows were experimentally infected with bovine viral diarrhoea virus (BVDV) between day 74 and 81 of pregnancy. All cows became infected and developed serum antibodies. Sixteen of the cows delivered persistently infected (PI) offspring, whereas the remaining three gave birth to calves with detectable serum antibodies and free from BVDV. The 16 cows with PI foetuses developed higher levels of antibodies in serum during pregnancy than did their three peers carrying non-PI calves. Multivariate analysis showed that the antibody levels in these two groups of cows were significantly different from day 135 of pregnancy. Foetal fluid was successfully collected from 18 of the 19 infected cows and from five uninfected control cows between 10 and 24 days before delivery by use of a percutaneous, blind puncture technique. No negative effects were observed in the cows or their offspring. BVDV was isolated and detected with an immunoperoxidase test in foetal fluid from 13 of the 16 cows carrying PI foetuses, and from 15 of the cows when a quantitative fluorescent polymerase chain reaction (PCR) technique was used. The negative sample in the PCR assay was positive for BVDV antibodies. The number of viral copies per microlitre in foetal fluids varied between 103 and 1080 in the positive samples. All samples taken from the cows carrying non-PI foetuses were negative for BVDV in both assays. In this experiment, examination of either serum or foetal fluids could identify the cows carrying a PI foetus. Examination of serum for BVDV antibodies was a reliable indicator of a PI foetus if the serum was collected during the last 2 months of pregnancy. For examination of foetal fluids, both viral and serological analyses should be performed. For viral analysis, PCR should be the test of choice. High levels of BVDV antibodies in conjunction with a negative result in the PCR may be indicative of a false-negative virus result. Further experience with the method of collection of foetal fluids is necessary for evaluation of its safety. Investigation of pregnant cows in order to discover a PI offspring before it is born could be a useful tool in control and eradication of BVDV.     

Embryo transplantation in cattle. Non-surgical transfer of 6 1/2--7 1/2 day old embryos to lactating dairy cows under farm conditions.

Previous attempts to transfer embryos non-surgically to heifers have been rather discouraging. The present experiment describes a simple non-surgical technique where the 6½–7½ -day old embryo is placed in a 0.25 ml ministraw, fitted into a common insemination gun and transferred cervically to synchronized lactating dairy cows. Thirty-two viable embryos were transferred to the middle part of the uterine horn ipsilateral to the ovary containing the corpus luteum. It was easy to pass the cervix, and most embryos could be deposited in the horn within 1 or 2 min. Eighteen animals were diagnosed pregnant (56% ). It is conceivable that the easy atraumatic transfers and good management (feeding, oestrus control) of the recipients contributed to the high pregnancy rate.     

Serum Profiles of Pregnancy-Associated Glycoprotein, Oestrone Sulphate and Progesterone During Gestation and Some Factors Influencing the Profiles in Ethiopian Borana and Crossbred Cattle

This study presents serum concentrations of pregnancy‐associated glycoprotein (PAG), oestrone sulphate (E1‐S) and progesterone (P4), and the effects of some dam and foetus‐related factors on these profiles during gestation in Borana and crossbred cattle. The PAG concentrations at 4th week post‐conception ranged from 1.5–5.5 and 2.1–4.7 ng/ml in Borana (n = 6) and crossbred (n = 8) cattle, respectively. The mean PAG concentrations increased progressively from 4th to 33rd week of gestation (from 3.3–173 ng/ml for Borana and 4.2–240 ng/ml for crossbred cattle) and reached peak around calving. Breed, parity status, dam body weight, foetal sex and foetal birth weight significantly influenced the PAG concentrations. After delivery, the PAG concentrations declined steadily to 5.7 ng/ml in Borana (n = 7) and 3.9 ng/ml in crossbred (n = 6) cattle 10 weeks post‐partum. The serum E1‐S concentrations at 17th week of pregnancy ranged from 0.3–2.6 and 0.9–5.7 ng/ml in Borana (n = 8) and crossbred (n = 9) cattle, respectively. The mean E1‐S concentrations increased progressively from 17th to 33rd week of gestation (from 1.1–4.6 ng/ml for Borana and 2.7–10.8 ng/ml for crossbred). Breed, parity status, dam body weight and foetal sex significantly influenced E1‐S concentrations. The P4 concentrations at 4th week of pregnancy ranged from 3.2–5.1 and 1.7–8.9 ng/ml in Borana (n = 6) and crossbred (n = 8) cattle, respectively. The P4 level remained elevated throughout pregnancy. This study indicated that the serum PAG and P4 concentrations at 4th and E1‐S approximately 17th week of pregnancy were above the cut‐off value for pregnancy test and the hormonal profiles observed were comparable to the previous reports. Furthermore, the PAG and E1‐S profiles were considerably influenced by factors such as breed, weight and parity status of the dam, and foetal sex and foetal birth weight (only PAG).     

Use of insulin-like growth factor-I during embryo culture and treatment of recipients with gonadotropin-releasing hormone to increase pregnancy rates following the transfer of in vitro-produced embryos to heat-stressed,lactating cows

An experiment was conducted to determine whether pregnancy rates following the transfer of in vitro-produced embryos to heat-stressed cows could be improved by 1) culturing embryos in the presence of IGF-I and 2) treating recipients with GnRH. Lactating Holstein cows (n = 260) were synchronized using a timed ovulation protocol. Embryos were produced in vitro and cultured with or without 100 ng/mL of IGF-I. On d 7 after anticipated ovulation (d 0), a single embryo was transferred to all recipients with a palpable corpus luteum (n = 210). A subset of recipients (n = 164) was injected with either GnRH or placebo on d 11. Plasma progesterone concentrations on d 0 and 7 were used to determine the synchrony of recipients. Pregnancy was diagnosed at d 53 and 81 by rectal palpation. Among all recipients, transfer of IGF-I-treated embryos increased pregnancy rate at d 53 (P < 0.05) and tended to increase pregnancy rate at d 81 (P < 0.06). Calving rate also tended to be higher for recipients that received IGF-I-treated embryos (P < 0.07). Among the subset of synchronized recipients (n = 190), pregnancy rate at d 53 and d 81 and calving rate were higher (P < 0.05) for IGF-I-treated embryos. The GnRH tended to increase pregnancy rate at d 53 for all recipients (P < 0.08) and the subset of synchronized recipients (P < 0.10). There were no effects of GnRH (P > 0.10) for pregnancy rate at d 81 and calving rate. The overall proportion of male calves was 64.3%. There was no effect (P > 0.10) of embryo treatment or GnRH on the birth weight or sex ratio of calves. Results of this experiment indicate that treatment of embryos with IGF-I can improve pregnancy and calving rates following transfer of in vitro-produced embryos. Further research is necessary to determine whether the treatment of recipients with GnRH is a practical approach to increase pregnancy rates following in vitro embryo transfer.     

Corpus Luteum–Endometrium–Embryo Interactions in the Dairy Cow: Underlying Mechanisms and Clinical Relevance

Conception rates of dairy cows are currently declining at an estimated 1% every year. Approximately, 35% of embryos fail to prevent luteolysis during the first three weeks of gestation. Interactions between the corpus luteum, endometrium and embryo are critical to the successful establishment of pregnancy and inadequacies will result in the mortality of the embryo. For example, as little as a one day delay in the post-ovulatory rise of progesterone has serious consequences for embryo development and survival. Recently, we found that LH support, degree of vascularization and luteal cell steroidogenic capacity were not the major factors responsible for this luteal inadequacy, but are nevertheless essential for luteal development and function. Progesterone acting on its receptor in the endometrium stimulates the production of endometrial secretions on which the free-living embryo is dependent. However, their exact composition and effects of inadequate progesterone remains to be determined. The embryo is recognized through its secretion of interferon tau (IFNT), which suppresses luteolytic pulses of prostaglandin F_2α. In the cow, it is most likely that IFNT inhibits oxytocin receptor up-regulation directly and does not require the prior inhibition of oestrogen receptor α (ESR1). Unravelling the precise luteal-endometrium and embryo interactions is essential for us to understand pregnancy establishment and development of strategies to reverse the declining fertility of dairy cows.     

Factors affecting plasma prolactin concentrations throughout gestation in high producing dairy cows

The aim of the present study was to investigate possible relationships between plasma concentrations of prolactin and the following factors throughout gestation in lactating dairy cows: photoperiod, season, milk production, Neospora caninum-seropositivity, twin pregnancy, and plasma concentrations of pregnancy associated glycoproteins-1 (PAG-1), progesterone and cortisol. Pregnancy was diagnosed by trans-rectal ultrasonography on Day 40 post-insemination and by palpation per rectum on Days 90, 120, 150, 180 and 210 or until abortion in aborting cows. Blood samples were collected from each animal immediately before each pregnancy diagnosis. The study population was comprised of 73 non-aborting (54 Neospora-seropositive cows) and 20 aborting cows (all Neospora-seropositive) cows. In non-aborting cows, GLM repeated measures of analysis of variance revealed that lactation number and days in milk had no effect on plasma prolactin concentrations throughout gestation, whereas high plasma prolactin concentrations were significantly associated with high plasma levels of cortisol and PAG-1, with Neospora-seropositivity, positive photoperiod (increasing day length), twin pregnancy, and with low plasma progesterone concentrations. An interaction among plasma prolactin, plasma cortisol and milk production was observed in that plasma prolactin concentrations differed significantly throughout gestation and were highest in high-producing cows with high cortisol levels. In Neospora-seropositive non-aborting versus aborting cows, mean prolactin concentrations failed to differ on Days 40, 90, 120, 150 and 180 of pregnancy, whereas prolactin values were significantly lower (P = 0.03) in aborting animals on Day 210 of pregnancy. Our results indicate that a positive photoperiod and Neospora-infection lead to increased plasma prolactin concentrations throughout gestation. Reduced prolactin concentrations detected in Neospora-seropositive aborting cows compared to non-aborting animals suggests a protective effect of prolactin in N. caninum infection.     

Plasma Testosterone in Cows Related to Day of Gestation and Sex of the Foetus,and Plasma Testosterone Levels Around Parturition

No significant differences in plasma testosterone level were observed between cows carrying a male foetus and cows carrying a female foetus at any ten-day interval from day 35 of gestation until parturition. Reported higher abortion rates for male than for female foetuses would thus appear not to be due to effects of foetal testosterone on the maternal endocrine balance. In spite of a great individual variation in plasma testosterone values at similar stages of gestation, certain trends are evident. From the 35th to the 80th day of gestation the average concentration was 90–100 pg/ml. Later it rose and reached 200 pg/ml on the 180th day, remaining at this level until after partus. During the first day after parturition plasma testosterone fell significantly and stabilized around 120 pg/ml.     

Progesterone and equine chorionic gonadotropin concentrations around the time of pregnancy loss in mares impregnated by donkeys or stallions

The objective of this study was to compare the incidence of pregnancy loss of mares carrying a mule embryo with that of mares carrying a horse embryo. The possible causes of such mortality were evaluated through serial ultrasonographic evaluations and hormonal monitoring, paying special attention to the role of premature regression of the endometrial cups and its relation to inadequate luteal function.

Twenty-eight mares impregnated by stallions and 19 mares impregnated with donkey semen were evaluated ultrasonographically every week from day 20 to day 150 of pregnancy. The viability of the product was assessed each time, and the diameter of the embryonic vesicle was measured from day 25 to day 60. Blood samples for progesterone and equine chorionic gonadotropin (eCG) determination were taken every week.

Both progesterone and eCG concentrations during normal pregnancies wegre significantly lower in the mares inseminated with donkey semen than in the mares impregnated by stallions (P<.05). In 7 of the mares carrying mule conceptuses to term, the concentrations of eCG remained basal throughout the study. In the other animals from this group, the levels of this hormone did increase but returned to baseline much earlier (on day 77 of pregnancy) than in the mares served by stallions (on day 126 day of pregnancy). There was no significant difference between the growth rate of embryonic vesicles of mares carrying mule embryos and that of mares carrying horse embryos (P>.05).

The incidence of pregnancy loss was significantly higher (P<.05) in mares carrying a mule embryo (36.8 %) than in mares carrying a horse embryo (21.4%); it occurred on average on day 93 of pregnancy in mares carrying mule embryos and on day 43 on mares carrying horse embryos. There was only 1 case in which pregnancy loss was associated with concentrations of both eCG and progesterone that were much lower than the average for the normal pregnancies of the same group, and this was in a mare carrying a horse embryo. The most frequent cause of pregnancy loss was premature luteal regression due to primary luteolysis, as evaluated via peripheral progesterone concentrations. This occurred in 2 mares carrying horse embryos and in 4 mares carrying mule embryos. Three mares carrying mule embryos and 1 carrying a horse embryo had abortions that were not preceded or accompanied by any alteration in progesterone or eCG levels and were thus classified as fetal deaths of non-endocrine origin.

It is concluded that the incidence of pregnancy loss is higher in mares carrying a mule embryo than in mares carrying a horse embryo. However, this is not due to the low progesterone concentrations associated with the premature regression of the endometrial cups that occurs in mares with interspecific pregnancy.     

Ultrasonographic study of early embryonic loss induced by Actinomyces pyogenes in cattle.

Four cows were inoculated into the uterus with Actinomyces pyogenes between 30 and 41 days of gestation. Gross morphological changes were monitored by rectal palpation and with a realtime B-mode two-dimensional scanner with a 7.5 MHz transrectal linear transducer, shortly before infection and afterwards at three to 12 hours intervals. Two control groups of cows 27 to 50 days pregnant were used: two cows were inoculated with 6 ml of sterile saline into the uterine lumen and four cows were treated with cloprostenol (prostaglandin F2 alpha analogue). There was a change in the uterine fluid from a black, non-echogenic image before infection to a grey or cloudy echogenic image as early as 21 hours after infection. There was an increase in the thickness of the endometrium of the horns and body of the uterus. The embryonic membranes thickened and separated from the endometrium as early as four hours after infection, followed by cessation of the embryonic heart beat, opening of the cervix and abortion. Abortion was followed by an increase in the amount of echogenic intrauterine fluid leading to an increase in the size of the uterus, and the cervix remained open for at least eight days. The area of the corpus luteum remained greater than 2 cm2 throughout the whole period. Intrauterine inoculation with sterile saline had no effects, but the administration of cloprostenol was followed by the death of the embryo and abortion within 72 hours, and the regression of the corpus luteum from greater than 2 cm2 at treatment to 0.4 cm2, 24 hours after abortion.(ABSTRACT TRUNCATED AT 250 WORDS)