Ultrasonic detection of the conceptus and characterization of intrauterine fluid on days 10 to 22 in heifers

Nonbred and pregnant heifers were examined ultrasonically on alternate days from Days 10 to 22 in three experiments. The goal was to define criteria that could be used for identification of the early conceptus. The location and size of circular and elongated nonechongenic structure (apparently free intrauterine luminal fluid) were recorded (Experiment 1). On Day 12, there was a significantly greater number of circular nonechogenic structures in the ovarian half of the uterine horn ipsilateral to the corpus luteum in pregnant heifers than in nonbred heifers (means, 5.1 vs 3.5). Therefore, the embryonic vesicle was apparently detected on Day 12, but was not distinguished from circular nonechogenic structures representing free luminal fluid. The number of circular nonechogenic structures decreased after Day 14, with a corresponding increase in elongate nonechogenic structures associated with luteal regression (nonbred heifers) or elongation of the blastocyst (pregnant heifers). On Day 16, there were more elongated than circular nonechogenic structures in pregnant heifers than in nonbred heifers. In nonbred heifers, there was no evidence of a local utero-ovarian relationship between the amount of intrauterine luminal fluid and the location of either the corpus luteum or the next ovulatory follicle. Uterine luminal fluid in each horn increased several fold between Days 16 and 18 in nonbred heifers; there was significantly more lumina fluid in nonbred than in pregnant heifers on Days 18 and 20. In pregnant heifers, intrauterine luminal fluid area increased first in the ovarian end of the uterine horn ipsilateral to the corpus luteum; this was apparently due to detection of the elongating blastocyst. The length of the longest specular reflector on the endometrial surface of the uterine lumen increased over Days 10 to 22 in nonbred and pregnant heifers, but was not a useful criterion for detection of the conceptus (Experiment 2). The accuracy of a 7.5 MHz transducer for pregnancy diagnosis was not greater than a guess (50%) before Day 16 (Experiment 3). Results indicated that early pregnancy diagnosis in heifers was confounded by the presence of intrauterine fluid.     

Mobility of twin embryonic vesicles in mares

The patterns of intrauterine mobility and fixation of multiple embryos were studied by ultrasonography in 33 mares with twins, six mares with more than two embryos, and 18 mares with singletons. For both single and multiple embryos, the embryonic vesicles showed a preference for the uterine body on days 11 (57% in body) and 12 (58%) and for the uterine horns on days 13 (40% in body), 14 (20%), 15 (15%), and 16 (1%). The preference for the uterine body was characteristic of vesicles that were 3 to 9 mm in diameter. Based on the number of individual embryos which were in different locations between two successive daily examinations, fixation (cessation of mobility) occurred for 97% of the embryos by day 16 and for all embryos by day 18. For 33 mares with twins, fixation involved one uterine horn in 23 mares and both horns in 10 mares (significantly different from equality). Location determinations were made every five minutes during two-hour trials on days 12, 13, or 14 in nine mares with singletons and ten mares with twins. Individual embryos of twin sets had mobility patterns similar to those of singletons. Summed over singletons and twins, the vesicles moved from one horn to another a mean of 0.9 times per two-hour trial (equivalent to 11 times per day). The smaller embryo of twin sets on the average spent more time in the uterine body, but this finding was attributed to their smaller diameter. The observed frequency with which both embryos of twin sets were simultaneously in a given segment of the uterus (28%) was greater (P<0.01) than the expected frequency if each embryo moved independently of the other (18%). Results indicated that 64% of the location changes of twin embryos occurred independently of one another, supporting the hypothesis that the embryonic vesicle plays an active role in its mobility.     

Spontaneous embryonic death on Days 20 to 40 in heifers

Transrectal ultrasound examinations were used in nulliparous Holstein heifers to study the association between time of spontaneous embryonic death (cessation of heartbeat) and luteal regression, and to determine the fate of the conceptus after embryonic death. There was no significant difference between nonbred heifers (n = 135) and bred, nonpregnant heifers (embryonic heartbeat never detected, n = 40) for day of onset of luteal regression (means, 17.6 and 17.9, respectively) or for length of interovulatory interval (means, 20.6 and 20.9 days, respectively). Pregnancy was confirmed by detection of an embryonic heartbeat on Day 24 (ovulation = Day 0) or later, or on two consecutive days prior to Day 24; on average, an embryonic heartbeat was detected on Day 22.0 (n = 104). Pregnancy rate on Day 24 was higher (P<0.02) in heifers bred on Day -1 (116/149, 77.8%) than in heifers bred on Day -2 (51/79, 64.6%), and was higher (P<0.05) in heifers with an embryo transferred ipsilateral to the corpus luteum than in heifers with an embryo transferred contralateral to the corpus luteum (3/4 vs 0/5). Embryonic death (lack of embryonic heartbeat following confirmation of pregnancy) and presumptive embryonic death (embryonic heartbeat detected on one day only, prior to Day 24) were detected prior to Day 25 in one and two bred heifers, respectively, and in one and two heifers with an embryo transferred contralateral to the corpus luteum, respectively. In these six heifers, luteal regression preceded, and apparently caused, embryonic death. In seven of eight heifers in which embryonic death was detected between Days 25 and 40, the onset of luteal regression was detected at least 3 d (range, 3 to 42 d) after detection of embryonic death. The incidence of embryonic death on Days 29 to 32 was lower (P<0.02) in heifers bred on Day -1 than in heifers bred on Day -2 (0 of 96 vs 3 of 40, respectively). In heifers in which luteal regression preceded embryonic death, the conceptus was lost rapidly, with minimal evidence of degeneration. In heifers in which embryonic death preceded luteal regression, there was ultrasonic evidence of conceptus degeneration, but conceptus fluid and tissue appeared to be maintained. In all heifers with embryonic death, the conceptus and its breakdown products apparently were eliminated by expulsion through the cervix rather than by resorption.     

Characterization of intrauterine mobility of the early equine conceptus

Intrauterine mobility patterns of the embryonic vesicle were characterized on Days 9 to 17 after ovulation in pony mares using real-time ultrasonography (n=5 or 7 mares per day). The location of the vesicle was determined by dividing the uterus into right horn, left horn, and body. Each uterine horn was further divided into three approximately equal portions (cranial third, middle third, caudal third), yielding seven segments (body plus three portions of each horn). Location of the vesicle within the uterus was recorded every five minutes for two consecutive hours (25 location determinations per trial). The number of times the vesicle was found in the uterine body versus one of the uterine horns was greater for the body on Day 9 (15.2 vs 9.8; not significant) and Day 10 (17.3 vs 7.7 P<0.05) and greater (P<0.05) for the horns on Days 12 (7.3 vs 17.7) through 17 (0.0 vs 25.0). Averaged over all days, when the vesicle was in one of the uterine horns it was present 56% of the time in the caudal third, 30% of the time in the middle third, and 14% of the time in the cranial third. Mobility was determined by the number of times the vesicle changed locations during successive examinations. On Day 9, the mean number of location changes per trial was minimal (horn to horn, 0.2; body to horn or vice versa, 1.8; between two segments, 4.2). The extent of mobility increased on Day 10 and reached an apparent plateau from Day 11 to Day 14. The mean number of location changes per trial during the plateau was as follows: horn to horn, 1.6; body to horn or vice versa, 5.6; between two segments, 10.7. Fixation (cessation of mobility) occurred in one of the horns in 5 7 mares on Day 15 and in 7 7 mares by Day 16. Mobility was present on the earliest day the embryonic vesicle was detected (Day 9), but Days 11 to 14 were characterized as the days of maximum mobility.     

Determination of early pregnancy and embryonic growth in goats by transrectal ultrasound scanning

Ultrasonography has been shown to be a useful tool for pregnancy diagnosis and the study of embryonic growth in mammals. The objectives of this study were 1) to evaluate the use of real-time B-mode ultrasonography for early pregnancy diagnosis in goats, 2) to define criteria for accurate diagnosis of pregnancy, and 3) to monitor the embryonic growth ultrasonically until Day 40 after mating. Estrus was synchronized in 16 cyclic Anglo-Nubian goats with a single injection of cloprostenol (125 micrograms, i.m.). Estrous females were randomly assigned into 2 groups: 1) goats mated by a vasectomized male (n = 5; MV group), and 2) goats mated by an intact male of proven fertility (n = 11; MF group). Transrectal ultrasonographic examinations with a 5 MHz linear array transducer were performed from Days 13 to 40 post mating. The evaluated parameters included the appearance of nonechogenic areas in the uterus, presence of embryo(s), crown-rump length of embryo and embryonic heart rate (beats/min). On Day 18, the mean (+/- SEM) diameter of nonechogenic areas was 1.5 +/- 0.3 mm in the MV group and 4.0 +/- 0.5 mm in the MF group (P < 0.01). In 36% of the pregnant does these areas were less than 3 mm. The mean (+/- SEM) day of the first detection by means of heartbeats of at least 1 embryo was 20.7 +/- 0.5 d (range, Days 19 to 23). From Days 19 to 38 of pregnancy, crown-rump length was best represented by a linear regression (Y = -2.23 + 0.13X; r2 = 0.94; P < 0.05). Crown-rump length on the day of the first detection of an embryo was 5.3 +/- 0.3 mm, reaching 34.2 +/- 0.6 mm on Day 40. Mean (+/- SEM) heartbeat rate was 168.3 +/- 2.8 beats/min on Day 21, decreasing to 158.3 +/- 2.0 beats/min on Day 40. Detection of the caprine embryo by ultrasonography and confirmation of its viability by heartbeats was shown to be a reliable method for early pregnancy diagnosis in Anglo-Nubian goats. Ultrasonic measurement of crown-rump length was useful in predicting the age of the embryo.     

Collection of tubal stage bovine embryos by means of endoscopy. A technique report

Here we describe the development and optimization of endoscopy-mediated transvaginal access for collecting ova and embryos from the bovine oviduct. The novel technique was developed in three experimental setups: In Experiment 1 embryos were collected unilaterally from nonstimulated heifers. We flushed the oviducts of superovulated heifers unilaterally (Experiment 2) and bilaterally (Experiment 3). In Experiment 1 the oviducts of 18 heifers were successfully cannulated, which resulted in the collection of twelve 1-cell to 8-cell embryos and one empty zona. Unilateral flushing of 13 animals (Experiment 2) resulted in 84 ova with 6.3 +/- 3.2 observed ovulation sites. Bilateral flushing of 25 animals (Experiment 3) resulted in 293 ova plus 10 empty zonae from 11.8 +/- 5.4 ovulation sites. Given our experience from these studies we optimized the technical equipment by improving the flushing metal catheter (Experiment 4). The novel catheter hermetically sealed the lumen of the ampulla at the moment, the medium was flushed through the oviduct. This resulted in a visible flow of medium via oviducts toward the embryo filter connected to an embryo flushing catheter that was fixed in the uterine horns. Our endoscopy-guided method is minimally invasive and facilitates the flushing of tubal stage embryos.     

Unilateral uteroovarian relationship in pregnant cattle and role of uterine vein

The hypothesis of a unilateral uteroovarian relationship between location of embryos and maintenance of corpora lutea (CL) was tested in superovulated cattle. Ovulations were induced in both ovaries and a uterine horn was isolated subsequent to insemination to produce unilateral pregnancy. The mean weight of CL on day 24 was greater (P<.05) on the gravid side (2580 mg) than on the nongravid side (1200 mg; n=5), supporting the hypothesis. Involvement of the main uterine vein in the unilateral pathway between a gravid horn and the adjacent ovary was demonstrated in an experiment which utilized separation of uterine horns, anastomosis of uterine veins between sides, and insertion of embryos into one side by embryo transfer. Mean weight of CL on day 24 was less (P<.05) when the gravid horn was contralateral to the CL (950 mg; n=3), than when the gravid horn was ipsilateral to the CL (4760 mg) or when the gravid horn was contralateral to the CL and the uterine vein from the contralateral side was anastomosed to the vein on the ipsilateral side (3580 mg; n=3).     

Changes in vascular perfusion of the endometrium in association with changes in location of the embryonic vesicle in mares

The equine embryonic vesicle is mobile on Days 12-14 (Day 0 = ovulation), when it is approximately 9-15 mm in diameter. Movement from one uterine horn to another occurs, on average, approximately 0.5 times per hour. Mobility ceases (fixation) on Days 15-17. Transrectal color Doppler ultrasonography was used to study the relationship of embryo mobility (experiment 1) and fixation (experiment 2) to endometrial vascular perfusion. In experiment 1, mares were bred and examined daily from Day 1 to Day 16 and were assigned, retrospectively, to a group in which an embryo was detected (pregnant mares; n = 16) or not detected (n = 8) by Day 12. Endometrial vascularity (scored 1-4, for none to maximal, respectively) did not differ on Days 1-8 between groups or between the sides with and without the corpus luteum. Endometrial vascularity scores were higher (P < 0.05) on Days 12-16 in both horns of pregnant mares compared to mares with no embryo. In pregnant mares, the scores increased (P < 0.05) between Day 10 and Day 12 in the horn with the embryo and were higher (P < 0.05) than scores in the opposite horn on Days 12-15. In experiment 2, 14 pregnant mares were examined from Day 13 to 6 days after fixation. Endometrial vascularity scores and number of colored pixels per cross-section of endometrium were greater (P < 0.05) in the endometrium surrounding the fixed vesicle than in the middle portion of the horn of fixation. Results supported the hypothesis that transient changes in endometrial vascular perfusion accompany the embryonic vesicle as the vesicle changes location during embryo mobility.     

The position effect in mice on day 14

Previous publications have produced conflicting results concerning the effect and nature of uterine horn position on embryonic weight and development in the uterus bicornis. For this investigation we used 503 embryos from 41 litters of NMRI - mice on gestational day 14. The relative position of each embryo was determined by measuring the distance from the center of each placenta to the cervix in relation to the total length of the uterine horn. After cessation of shrinkage due to fixation the umbilical cord and remaining membranes were removed under the dissecting microscope and the embryos were weighed to the nearest 0.1 mg. Analysis of potential embryonic positions along the uterine horns indicated that 10% at the extreme ovarian and cervical ends were almost devoid of implantations. A mixed nested analysis of variance was used as the appropriate model to test a position on weight effect. Not only was a position effect found but, in contrast to other studies, it is also quantitatively represented. Embryos were lighter both at the ovarian and cervical ends, the mean weight difference being in the range of less than 10% of the average embryo weight.     

Mobility of the early equine conceptus

Movement of the conceptus within the uterine lumen of barren mares was studied by daily ultrasound examinations on days 11-20 and by rectal palpation on days 15-48 (Experiment 1) and by ultrasound examinations 3 or 4 times per day at 2-4 hour intervals on days 11-16 (Experiment 2). In addition, broodfarm records were analyzed to compare side of ovulation with side of embryo attachment (Experiment 3). The vesicle was found in opposite uterine horns for 43% of the successive, daily, ultrasound examinations on days 11 and 12, 12 and 13, 13 and 14, and 14 and 15; 24% of the successive examinations on days 15 and 16; and 8% on days 16 and 17. No movement was detected after day 17. The vesicle was found in opposite horns during 41% of the successive examinations at 2-4 hour intervals on days 11, 12, 13, 14, and 15, but no movement was detected on day 16. In addition, no transuterine migration was found by rectal palpation between the day of first detection of an embryonal enlargement (mean, day 17) and day 48. During ultrasound examination on days 11-15, the vesicle was found significantly more frequently in the left horn (66% of the observations) than in the right horn (34%); however, final attachment occurred more frequently in the right horn (63% of the mares). In analyses of brood-farm records, ovulation occurred with equal frequency in left and right ovaries in barren and lactating mares, but with significantly greater frequency in the left ovary (63%) in maiden mares. Regardless of the side of ovulation, final attachment of the conceptus occurred significantly more frequently in the right horn (66%) in barren and maiden mares, but not in lactating mares.     

Effect of transfer of one or two in vitro-produced embryos and post-transfer administration of gonadotropin releasing hormone on pregnancy rates of heat-stressed dairy cattle

Pregnancy rates following transfer of an in vitro-produced (IVP) embryo are often lower than those obtained following transfer of an embryo produced by superovulation. The purpose of the current pair of experiments was to examine two strategies for increasing pregnancy rates in heat stressed, dairy recipients receiving an IVP embryo. One method was to transfer two embryos into the uterine horn ipsilateral to the CL, whereas the other method involved injection of GnRH at Day 11 after the anticipated day of ovulation. In Experiment 1, 32 virgin crossbred heifers and 26 lactating crossbred cows were prepared for timed embryo transfer by being subjected to a timed ovulation protocol. Those having a palpable CL were randomly selected to receive one (n = 31 recipients) or two (n = 27 recipients) embryos on Day 7 after anticipated ovulation. At Day 64 of gestation, the pregnancy rate tended to be higher (P = 0.07) for cows than for heifers. Heifers that received one embryo tended to have a higher pregnancy rate than those that received two embryos (41% versus 20%, respectively) while there was no difference in pregnancy rate for cows that received one or two embryos (57% versus 50%, respectively). Pregnancy loss between Day 64 and 127 only occurred for cows that received two embryos (pregnancy rate at Day 127=17%). Between Day 127 and term, one animal (a cow with a single embryo) lost its pregnancy. There was no difference in pregnancy rates at Day 127 or calving rates between cows and heifers, but females that received two embryos had lower Day-127 pregnancy rates and calving rates than females that received one embryo (P < 0.03). Of the females receiving two embryos that calved, 2 of 5 gave birth to twins. For Experiment 2, 87 multiparous, late lactation, nonpregnant Holstein cows were synchronized for timed embryo transfer as in Experiment 1. Cows received a single embryo in the uterine horn ipsilateral to the ovary containing the CL and received either 100 microg GnRH or vehicle at Day 11 after anticipated ovulation (i.e. 4 days after embryo transfer). There was no difference in pregnancy rate for cows that received the GnRH or vehicle treatment (18% versus 17%, respectively). In conclusion, neither unilateral transfer of two embryos nor administration of GnRH at Day 11 after anticipated ovulation improved pregnancy rates of dairy cattle exposed to heat stress.     

Improvement in recovery of embryos/ova using a shallow uterine horn flushing technique in superovulated Holstein heifers

The aim of this study was two-fold: (1). to compare recovery of embryos/ova from superovulated Holstein heifers by flushing the uterine horns through insertion of the catheter very close to the tip of the horn (deep) or just after the uterine bifurcation (shallow) and (2). to evaluate the hormonal and superovulatory response to estradiol benzoate (EB) treatment prior to superovulation. Ten Holstein heifers (12-16 months) underwent two superovulatory treatments in a cross-over design. Heifers were treated with decreasing doses of FSH from Days 8 to 12.5 of a synchronized estrous cycle. At 4 days prior to superovulation, half of the heifers received EB (5mg, i.m.) or served as Controls, followed by the alternative treatment in the subsequent superovulation. At embryo recovery, one uterine horn was flushed with deep ( approximately 7 cm caudal to the tip of the horn) and the other with shallow ( approximately 5 cm cranial to the beginning of the uterine bifurcation) flushing techniques. Embryos/ova were recovered, counted, and scored. Number of ovulations was estimated by ultrasound. Pretreatment with EB reduced circulating FSH and regressed the first wave dominant follicle with no change in number of large follicles, number of ovulations, number of embryos/ova recovered, or number of transferable embryos. The shallow flushing technique was superior to the deep technique for number of embryos/ova recovered per horn (5.4+/-1.1 versus 3.9+/-0.8) or percentage of embryos/ova recovered per CL (63.9+/-8.6% versus 37.4+/-6.5%). Thus, flushing the entire uterine horn increased recovery of embryos/ova.     

Autotransfer of Day 4 embryos from oviduct to oviduct versus oviduct to uterus in the mare

Embryo autotransfer is defined as the collection of an embryo from and the transfer of this embryo into the same animal. The objectives of this study were to: 1) test the hypothesis that oviduct transport of the equine embryo from the oviduct into the uterus is not dependent on a unilateral embryo-corpus luteum interaction, 2) develop an embryo autotransfer technique for the mare and 3) compare the success rates of Day 4 embryos surgically autotransferred from the oviduct ipsilateral to ovulation to either the oviduct (n=10 mares) or the uterine horn (n=10 mares) contralateral to ovulation. Seventy percent (7 10 ) of the Day 4 embryos which were autotransferred to the oviduct contralateral to ovulation were transported through the oviduct and subsequently developed into embryonic vesicles detectable by ultrasonography between 10 and 21 days postovulation. This finding supported the hypothesis that oviductal embryo transport is not dependent upon the ipsilateral corpus luteum. Overall, sixty percent (12 20 ) of the autotransfers were successful. The success rate of uterine-transferred embryos was not significantly less (P>0.3) than that of oviductal-transferred embryos (5 10 vs 7 10 , respectively). Therefore, the Day 4 equine embryos were apparently mature enough to survive in the mare's uterus.     

Fertilization rates and embryonic development in superovulated cattle inseminated in different sites within the reproductive tract

Superovulated Jersey and Holstein heifers and cows were bred 9.7 ± 2.7 h after the first observation of estrus with a single dose of frozen semen. Animals were grouped by site of insemination: 1) right uterine horn (n = 5), 2) left uterine horn (n = 4), 3) mid- uterine body (n = 5), and 4) mid-cervix (n = 6). The number of unfertilized ova, normal and abnormal embryos were recorded for each horn at slaughter 115.8 ± 18.6 h after insemination. All viable embryos were cultured in vitro and assessed for development. The overall fertilization rate was 76.8%, with the ipsilateral horn being higher than that of the contralateral horn (P < 0.05). Similar fertilization rates resulted among all treatments except those inseminated in the contralateral horn (P < 0.05). In vivo development of embryos was higher for the ipsilateral horn inseminations than those of the body of the uterus or cervical inseminations (P < 0.05), but it was not higher than the contralateral horn inseminations (P > 0.05). Under the conditions of this study with superovulated cows, these results suggest than 1) spermatozoa migrate from one horn to the other and 2) inseminating too deep into one horn may reduce the chances for concention when ovulation occurs contralaterally.     

Maintenance of the corpus luteum after uterine transfer of trophoblastic vesicles to cyclic cows and ewes

One or two trophoblastic vesicles (0.4-2 mm diam.) from cow (Day 14) or ewe (Day 11-13) embryos without their disc were transferred, after culture for 24 h, into recipients. Each vesicle was transferred into the uterine horn ipsilateral to the CL by the cervical route in heifers and surgically in ewes on Day 12 of the oestrous cycle. In cows, daily measurements of plasma progesterone concentrations and checks for return to oestrus showed that the CL was maintained in 8 out of 12 recipients. These 8 cows had 25- to 37-day cycles while 4 recipient heifers returned to oestrus normally. Three recipients with an extended cycle were slaughtered. The dissected uterus showed that trophoblastic vesicles had developed in the uterine horns. In ewes, the serum progesterone curve, determined in each recipient, showed that the CL was maintained in 7 out of 12 recipients. These 7 ewes had 20- to 54-day cycles and the other 5 ewes had a normal cycle of 15-19 days comparable to that of 17.0 +/- 0.5 days for the 6 control ewes. Whenever the CL was maintained, high blood progesterone levels were followed by rapid luteolysis. In cattle and sheep, therefore, a trophoblastic vesicle transferred into the uterus can develop in vivo, secreting the embryonic signals when there is no embryonic disc control and transforming the cyclic CL into a CL of pregnancy in about 60% of the cases.(ABSTRACT TRUNCATED AT 250 WORDS)     

Transuterine embryo migration in recipient cattle

Transuterine migration of bovine embryos following fertilization in vivo is apparently rare, but little is known about migration following embryo transfer. We studied heifers receiving either 1 or 2 in vitro produced embryos to determine 1) the incidence of transuterine migration, 2) the timing of migration and 3) the random or systematic occurrence of the event. In 4 experiments, 436 heifers received embryos and 218 of these were pregnant at necroscopy on either Day 14, Day 18, Day 26 or Day 60 of pregnancy. Overall, 43/218 (20%) of the heifers had embryos that had migrated. The frequency of migration was higher in twin (30/68) than in single (13/150) embryo transfers of pregnant recipients (44 vs 9%; P<0.001), and in contralateral (9/15) than in ipsilateral (33/170) transfers (60 vs 19%; P<0.001). Among the heifers that received embryos by ipsilateral transfer, the migration rate was similar to that in heifers pregnant with a singleton after the transfer of either 1 (2/48) or 2 (4/60) embryos (4 vs 7%, NS). The migration rate was highest at Day 26 (12/37) in heifers receiving twin embryos by ipsilateral transfer but was similar at all other stages of pregnancy (15/111, 32 vs 14%; P<0.01). Migration was first observed by Day 14, and it appears that either further migration occurred over the next 12 d or that migration was associated with a higher survival rate from Day 14 to Day 26. The low migration rate evident at Day 60 suggests that migration by Day 26 was associated with increased embryo or fetal death by Day 60. The data suggest that embryo migration is probably independent for each of a pair of surviving embryos. We conclude that in cattle embryo migration is embryo-dependent, but this capability is dormant unless more than 1 embryo is present in a uterine horn or the embryos are transferred to the contralateral uterine horn. The relationship between migration and embryo survival remains unclear.     

Uterine transport of prostaglandin E(2)-releasing simulated embryonic vesicles in mares

Transrectal ultrasonography was used to test the hypothesis that prostaglandin E(2) (PGE(2)) would increase the uterine transport of simulated embryonic vesicles in mares. Uterine transport of PGE(2)-releasing (PGE) vesicles, vehicle-releasing (sham) vesicles, and equine embryos was contrasted on Day 12 or Day 13 post ovulation. In Experiment 1, there was no difference (P>0.10) in transport of PGE vesicles, sham vesicles, Day-12 embryos, and Day-12 embryos after cervical manipulation (n = 3 per group). In Experiments 2 and 3, respectively, transport of PGE and sham vesicles was contrasted with transport of Day-13 embryos after the vesicles (1 vesicle per mare) were placed into the uterine lumen with the embryo, (n = 7 per group). In Experiment 2, PGE vesicles were transported less often (P<0.05) from horn to body and from segment to segment than Day-13 embryos before vesicle insertion. In Experiment 3, sham vesicles were transported less often from horn to body (P<0.10) and from segment to segment (P<0.01) than Day-13 embryos before vesicle insertion. However, there was no difference (P>0.10) in the transport of PGE vesicles and embryos (Experiment 2) or sham vesicles and embryos (Experiment 3) together in the uterine lumen. In Experiment 4, transport of PGE and sham vesicles was contrasted by placing them together into the uterine lumen of nonpregnant mares on Day 13 (n = 7). There was no difference (P>0.10) in the transport of PGE and sham vesicles together in the uterine lumen. These results do not support the hypothesis that PGE(2) increases uterine transport of simulated embryonic vesicles. In addition, these results do not support the hypothesis that equine embryos stimulate uterine transport.     

An early endometrial vascular indicator of completed orientation of the embryo and the role of dorsal endometrial encroachment in mares

The spherical equine embryonic vesicle is mobile throughout the uterine lumen for several days before becoming fixed in the caudal segment of a uterine horn on Day 16 (ovulation = Day 0). Orientation refers to the position of the embryo proper at the periphery of the vesicle relative to the position of the mesometrial attachment. In mares, the embryonic pole of the vesicle is antimesometrial after completion of orientation. Day of vesicle fixation, differential thickening of the endometrium near the mesometrial attachment, and orientation of the embryonic vesicle were studied in 30 ponies, using B-mode and color-Doppler transrectal ultrasonography. The thickness of the endometrium at the mesometrial aspect of the vesicle divided by the thickness at the antimesometrial aspect was termed the encroachment ratio. At the future site of fixation, the first increase (P < 0.05) in the encroachment ratio occurred between 4 and 1 days before fixation. An early vascular indicator of the future position of the embryo proper was discovered by color-Doppler imaging and consisted of a colored spot in the image of the endometrium close to the wall of the embryonic pole. The early indicator was detected in each mare 0.5 +/- 0.1 days after fixation and 2.5 +/- 0.2 days before first detection of the embryo proper. The position of the early indicator when first detected at the periphery of the embryonic vesicle was not significantly different from the position of the embryo proper when first detected. Results supported the hypothesis that differential thickening of the endometrium precedes orientation and indicated that orientation occurs immediately after fixation.     

Effect of a unilateral or bilateral twin embryo distribution on twinning and embryo survival rate in the cow

Uni- and bilateral twin embryo distributions were effected by the transfer of one embryo on Day 7 to the ipsi- or contralateral uterine horn of previously inseminated heifers (123, Exp. 1) or cows (95, Exp. 2). The embryo transfers were surgical in Exp. 1 and non-surgical in Exp. 2. Transferred and native embryos were distinguished by breed. Embryo survival rate was measured in a proportion (N = 40) of the heifers at Day 53 of gestation and in the remainder of the heifers and all of the cows at term. In the heifers (Exp. 1) overall pregnancy rates of 76% and 75% were recorded after uni- and bilateral twin embryo distributions respectively. Twinning rates of 55% and 60% at Day 53 of gestation and 60% and 60% at term were recorded for uni- and bilateral distributions respectively. In the cows (Exp. 2) calving rates of 61% and 63% and twinning rates of 33% and 38% were recorded following uni- and bilateral twin embryo distributions respectively. When the data from both experiments were combined, overall embryo survival rates were similar for both twin embryo distributions although the ipsilateral transfer of an embryo reduced the survival rate of the native embryo. It is concluded that the confinement of two embryos in one uterine horn on or after Day 7 does not depress pregnancy, twinning or overall survival rate to term.