Equine fetal kinetics: Presentation and location

The extent and nature of fetal mobility (presentation and location changes) were studied in 10 pony mares by ultrasonic examinations each week during the fetal stage (Day 40 to term). The percentage of examinations with cranial fetal presentation was 35 to 43% during Months 2 to 5 (no significant differences among months); a significant increase occurred between Months 5 and 6 reaching 100% at Month 7. With only 3 transient exceptions, the fetus was in cranial presentation during all examinations from Month 7 to term. The frequency of presentation changes between successive examinations decreased (P<0.05) between Months 4 and 5 and further decreased between Months 6 and 7. A significant and progressive decrease in the proportion of examinations in which at least part of the fetus was located in the umbilical-cord horn occurred over Months 2 to 4. During the same months, an increase occurred in the frequency of location of at least part of the fetus in the noncord horn and in the frequency of location of all of the fetus in the uterine body. The fetus was equally distributed among the 3 parts of the uterus during Month 4. Thereafter, locations decreased (P<0.05) in the noncord horn, increased (P<0.05) in the uterine body, and maintained a plateau (no significant differences) in the cord horn until Month 7 and then increased (P<0.05). The frequency of location changes between successive examinations increased (P<0.01) progressively during Months 2 to 4 followed by a decrease between Months 4 and 10. With one exception, the fetus was located in both the cord horn and uterine body for all examinations during Month 9 to term. In conclusion, a gradual decrease in fetal mobility occurred after Month 4; with only a few transient exceptions, final selection of cranial presentation occurred by Month 7, and final selection of partial location in the cord horn occurred by Month 9.     

Equine fetal kinetics: entry and retention of fetal hind limbs in a uterine horn

Transrectal ultrasonic examinations were made in 31 pregnant pony mares once a week during Months 6 to 11. Each uterine horn was divided into 3 approximately equal segments (caudal, middle, cranial). The percentage of examinations with cranial fetal presentation increased (P<0.05) progressively from 58% at Month 6 to 99% at Month 9 and was followed in all mares by entry of the fetal hind limbs into one uterine horn. The mean number of uterinehorn segments with limb parts increased (P<0.05) between each set of consecutive months from Month 6 to Month 10. Initially (Months 7 and 8), retraction of limbs after entry into the caudal and middle segments of the horn was common (31% incidence). The mean day of final entry of the limbs without detection of subsequent retraction was Day 230 +/- 2.2. Both uterine horns were closed during the examination preceding final entry of the hind limbs into one horn in 25 of 29 (86%) mares. The limbs reached the cranial segment in most examinations by Months 9 and 10 (73% and 98%). The cross-sectional height, as seen on the ultrasound screen, of both uterine horns increased (P<0.05) progressively during Months 7 to 10. Between Months 10 and 11, the height of the horn containing the limbs decreased (P<0.05); this result was attributable to a flattening of the horn and thinning (P<0.05) of the horn wall above a hoof and metatarsal bone. Apparent placental fluid (nonechogenic areas >5 mm in height) between the hind limbs and the cornual wall was detected in 0, 10, 25, and 63% (P<0.01) of examinations for Months 8 to 11, respectively. Results indicated that entry of the fetal hind limbs into a uterine horn was initially tentative and became final during a mean of Month 8. The apposition between the cornual wall and limbs was close during Months 7 to 11. By Month 11, the fetal-limb horn became flatter and thin-walled and usually contained placental fluid.     

Equine fetal kinetics: Allantoic-fluid shifts and uterine-horn closures

Uterine filling by the conceptus and uterine constrictions and allantoic-fluid shifts after filling were studied by daily ultrasound examinations on Days 40 to 74 in 10 mares. Allantoic fluid was first detected at the extremities of the uterus on the following days: 1) tip of umbilical-cord horn, Day 60.0 +/-1.9 (mean +/-SEM); 2) tip of noncord horn, Day 64.8 +/-1.3; and 3) near cervix, Day 48.5 +/-1.3. Transient uterine constrictions with expulsion of allantoic fluid were detected in 37% of 271 examinations. Other end points were studied by weekly ultrasound examinations from Day 40 to term. Height of cross-sections of the fetal body increased (P<0.01) progressively over Monthes 2 to 8, whereas the height of placental fluid minus fetal body decreased (P<0.05) after Month 3. An increasing (P<0.05) frequency of examinations with a reclosed uterine horn occurred between Months 3 and 4. The frequency of reclosed noncord horns continued to increase (P<0.05) after Month 4 and reached 96% at Month 7. The frequency of reclosure of cord horns plateaued (no significant differences) at 54% over Months 4 to 7 and then rapidly decreased (P<0.05). Both uterine horns were closed during the examination that preceded the permanent entry of the fetal hind limbs into the cord horn in all of 9 mares in which this temporal relationship was studied. After entry of the fetal hind limbs, the cord horn remained constricted upon the limbs until at least Month 10. The results provide rationale for the following hypotheses: 1) The gradual decrease in fetal mobility (location and presentation changes) previously reported for Months 4 to 8 is associated with growth of the fetus and a concomitant decrease in volume of allantoic fluid and number of open uterine segments; 2) Closure of both uterine horns near mid-pregnancy plays a role in final selection of cranial fetal presentation followed by entry of the fetal hind limbs into the cord horn.     

M-mode ultrasonic assessment of equine fetal heart rate

Embryonic heart rate was studied during 87 examinations in 56 mares on Days 25 to 174 post ovulation. Real-time B-mode using a stopwatch and M-mode ultrasonography were compared. For analyses of heart rates, data were grouped into weeks or months. For the stopwatch and M-mode techniques, respectively, heart rates averaged 123 and 133 beats per minute during Month 1, peaked during Month 3 at 172 or 196 beats, then gradually decreased over the duration of the study to 126 and 130 beats during Month 6. There was a significant increase in heart rate from Months 1 to 2 and Months 2 to 3 and a significant decrease from Months 3 to 4 and Months 5 to 6 for both techniques. There was a tendency for a difference (P<0.08) between techniques during Month 1 and a significant difference between techniques for each of Months 2 and 3. The lower heart rate values for the stopwatch technique during Months 1, 2 and 3 may have reflected the difficulties associated with maintaining the transducer field on the fetal heart continuously for an adequate period of time (Months 1 and 2) and difficulty counting fast enough to keep up with the fetal heart (Months 2 and 3). When data were grouped weekly, the largest increase in fetal heart rate occurred between Weeks 4 and 5.     

Ultrasonographic features of the mule embryo, fetus and fetal-placental unit

The aim of this study was to establish baseline ultrasound data concerning the mule conceptus during gestation. Ten multiparous Trotter mares were artificially inseminated with chilled semen from an Amiatino jack donkey. Daily transrectal ultrasonography was carried out from the day of ovulation until Day 50 of gestation to determine the following: first detection of the embryonic vesicle (EV), mobility phase, EV diameter, day of EV fixation, changes in EV shape, date of yolk sac regression and embryo crown-rump length. Monthly ultrasonic assessments from Day 50 of gestation to term were carried out. These assessments included an evaluation of fetal well-being and the growth of the mule conceptus, which were monitored using the following variables: cardiac activity, fetal activity and presentation, fetal fluid echogenicity, combined thickness of the utero-placenta unit and fetal orbital and aortic diameter. Mule EV first detection was observed earlier (37% at Day 8) than that observed in the equine pregnancy. EV diameter at first detection was 4.6 ± 1.1 mm. At Day 10, 75% of EVs were detected. EV fixation occurred on Day 17.1 ± 1.1, with a mean EV diameter of 2.5 ± 0.2 cm. EV growth rate was 4.04 mm/day from Days 11 to 16, 0.4 mm/day from Days 16 to 28 and 1.78 mm/day from Days 28 to 45 of pregnancy. The embryo proper was first detected on Day 19.9 ± 1.9 (average length 2.4 ± 1.4 mm), and the embryonic heartbeat was first detected on Day 24 ± 2.4. The fetal carotid pulse was observed at six months of gestation and provided a good means by which to estimate fetal cardiac activity in advanced gestation. The fetal heart rate was recorded from Month 2 of gestation to term. The mean ± SD of the combined uteroplacental thickness was assessed at the cervical-placental junction and at the ventral abdomen in mares between Months 2 and 5 until term, respectively. An abnormal fetal-placental unit and fetal inactivity was observed in association with abortion. Mule-conceptus biometric measurements correlated significantly with the gestational age, and these data were used to predict an unusually large mule fetus, which might result in dystocia. In conclusion, we can assume that early diagnosis of pregnancy failure and assessment of fetal biophysical profile and growth charts could improve the chances of gestation completion in mule-pregnant mares. The early detection of mares at risk for an abnormal pregnancy or delivery may increase the success of prompt treatments, therefore preventing costly emergency procedures and allowing proper obstetrical and neonatal assistance.     

Postfixation embryo reduction in unilateral and bilateral twins in mares

Multiple ovulations were induced with a pituitary extract in mares, and the development of multiple conceptuses was monitored daily by ultrasound on days 11 to 40. The incidence of abortion (loss of all embryos) was not significantly different between mares with multiple embryos (5 38 mares; 13%) and mares with singletons (4 36 ; 11%). Embryo reduction (elimination of excess embryos) was not detected during the embryo mobility phase (days 11-15) or on the day of fixation of embryos (day 16) in any of 38 mares with multiple embryos. The incidence of postfixation embryo reduction for mares with twins was 64% (18 28 ); however, the incidence for unilateral twins (17 19 ; 89%) was greater (P<0.01) than for bilateral twins (1 9 ; 11%). Reduction of unilateral twin embryos seemed to occur earlier (53% before day 20 and 82% before day 30) than for the set of bilateral twins (day 36). The remaining embryo in all mares in which embryo reduction occurred seemed normal in size and appearance on the last day of examination. However, in four of eighteen mares in which unilateral reduction occurred, the umbilical cord of the remaining embryo was attached in the ventral hemisphere of the all antochorion. This apparent disorientation was not seen in any of 16 bilaterally located embryos or in 16 singletons.     

The outcome of twin pregnancies in mares

Brood-farm veterinarians were surveyed and theriogenology records were examined to obtain information on the natural outcome of rectally-diagnosed twin pregnancies. Four of the veterinarians estimated that 50% of mares with twin embryos had single foals. In the analyses of palpation records, 31 53 (58%) of the diagnosed twin pregnancies terminated in birth of one foal. However, the loss of one embryo without the loss of the other occurred significantly more often before day 40-42 (30 53 ; 57%) than after day 40-42 (1 16 ; 6%). Of the remaining 15 mares with diagnosed twins at day 40-42, 5 had two foals and 10 had no foals. The methods used for intervention when twins were diagnosed were unsatisfactory. Complete termination of pregnancy with a prostaglandin or an intrauterine flushing resulted in failure to reestablish a singleton pregnancy during the operational breeding season in 10 11 mares. Attempts to eliminate one embryo resulted in loss of both in 6 7 mares. The results indicated that, at the present time, nonintervention should be given more consideration as a method of handling twin pregnancies.     

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.     

Ultrasonic fetal gender diagnoses during Months 5 to 11 in mares

The effect of fetal age on the ability to diagnose gender during the last half of pregnancy was evaluated in 85 mares using a 3.5 MHz linear-array transducer. Diagnosis of fetal gender was completed in 19 of 21 attempts (90%) during Months 5–6 and was independent of fetal presentation; the percentage of cranial, caudal, and transverse presentations was 52, 29 and 19%, respectively. Fetal gender was diagnosed in only 4 of 64 (6%) attempts during Months 7 to 11; fetal presentation was predominantly cranial (95%). Attempts at gender diagnoses were always successful when the fetus was in caudal or transverse presentation. Overall accuracy for fetal gender diagnoses was 89%. Absolute accuracy verified after birth of the foal was 100% when expected accuracy, estimated at the time of the examination, was ≥80%. During Month 5, the penis was a hyperechogenic trilobulated structure located near the fetal umbilical attachment. During Months 6 to 11, the penis was pendulous; it appeared circular in cross section and rectangular in longitudinal section. The anogenital raphe was a narrow hyperechogenic structure between the hind limbs and along the perineum. During Months 5 to 11, the clitoris was hyperechogenic, trilobulated, and located in the perineal region. The labia could be delineated when the entire perineum was imaged. The mammary gland was a triangular hyperechogenic structure between the hind limbs in frontal and cross-sectional planes. In both genders the anus was identified as a hyperechogenic structure located immediately below the tail. Ultrasonic fetal gender diagnosis was readily accomplished and most accurate during Months 5 to 6, but thereafter the diagnostic areas were usually inaccessible.     

Pregnancies, calves and calf viability after transfer of in vitro produced bovine embryos

Pregnancy, parturition and calf survival following the transfer of embryos produced in vitro were monitored. A total of 44 blastocysts was transferred in pairs to 1 uterine horn ipsilateral to the corpus luteum (CL) of 22 synchronized heifers. At Day 42 of development 14 recipients (64%) were pregnant; the calving rate was also 64%. The twinning rate was 9/14 at Day 42 and 7/14 at birth, for an overall fetal mortality rate of 9%. The average gestation length was 281 and 275 d for single and twin pregnancies, respectively. Blood samples from recipients were collected for determination of bovine pregnancy associated glycoprotein (bPAG) from 2 wk after transfer and throughout the pregnancy. During the first trimester of pregnancy, the bPAG concentration was significantly higher in twin than in single bearing heifers, and the perinatal increase in bPAG was correlated positively with the total weight of the fetus(es). The percentage of male calves was 43%. The birth weight of twin individuals was 25 +/- 1 kg, which was 78% of the birthweight of the singletons (32 +/- 2 kg). One singleton calf was oversized, weighing 58 kg (80% more than the median weight of the other singletons). Stillbirths occurred in 21% of the twins, butin none of the singletons. Calf mortality during the first 14 d was higher for twins (4/11) than for singletons (1/7) due to infections and cerebellar hypoplasia. Karyotyping the calves detected no cytogenetically recognizable abnormalities. All calves were negative for BVD virus and IBR antibodies. The results of this study showed that although the incidence of fetal loss was low, there was an unacceptable high perinatal mortality of the calves. Thus it is likely that the blood supply through the placenta of animals pregnant with twins was impaired or it is possible that these fetuses and calves had increased stress susceptibility caused by the in vitro conditions. Furthermore, the birth of 1 oversized calf, 2 calves with cerebellar hypoplasia and 5 calves succumbing to infections seems to indicate that a proportion of in vitro produced calves may suffer from factors inherent in the in vitro production system.     

Uterine contractile activity in mares during the estrous cycle and early pregnancy

Transrectal ultrasonography was used to quantitate uterine contractile activity during the estrous cycle and early pregnancy in pony mares (nonbred, n = 9; pregnant, n = 16). Continuous 1-min scans of longitudinal sections of the uterine body were videotaped, and uterine activity scores (1=minimal activity, 5=maximal activity) were assigned to each tape segment. There was a tendency (P<0.06) for a main effect of reproductive status (nonbred versus pregnant), a main effect of day (P<0.0001), and a reproductive status by day interaction (P<0.006). Uterine activity scores were higher (P<0.05) in pregnant mares on Days 1, 11, 12, and 17 (Day 0=day of ovulation) than in nonbred mares. Maximal activity in pregnant mares occurred on Days 11 to 14 during the reported period of maximal embryo mobility. Activity scores decreased (P<0.05) between the day prior to and the day of fixation (mean = Day 15) of the embryonic vesicle. Activity scores were maintained at an intermediate level for several days following fixation before declining to minimal levels by 7 d postfixation. A postovulatory decrease (P<0.04) in activity scores was observed in nonbred mares, but not in pregnant mares, between Days 0 and 1 followed by a progressive increase (P<0.03) between Days 2 and 4. Maximal activity in nonbred mares occurred during the late luteal phase (Days 13 to 14), corresponding temporally to the reported onset of luteolysis.     

Embryonic loss in mares: Nature of loss after experimental induction by ovariectomy or prostaglandin F(2alpha)

Twenty-one pregnant pony mares were assigned to one of the following groups: 1) controls, 2) ovariectomy at Day 12, 3) ovariectomy at Day 12 plus daily progesterone treatment on Days 12 to 40, 4) PGF(2alpha) on Day 12, 5) PGF(2alpha) on Day 21, and 6) PGF(2alpha) on Day 30. Based on daily examinations by ultrasound, the embryonic vesicle was maintained to Day 40 in all control mares and in mares that were ovariectomized on Day 12 and given progesterone. The embryonic vesicle was lost in all mares of the other four groups. Administration of progesterone prevented the embryonic loss associated with ovariectomy at Day 12, indicating that progesterone may be the only ovarian substance required for survival of the early embryo. The mean number of days to embryonic loss was greater for mares treated with PGF(2alpha) on Day 12 (6.8 days) than for mares ovariectomized on Day 12 (3.0 days). In the PGF(2alpha)-treated group, the vesicles did not become fixed at the expected time (Day 15), and mobility continued until the day of loss. In the mares treated with PGF(2alpha) on Day 21 and in one of the mares treated on Day 30, the vesicle was lost within one to three days without prior indication. Loss may have occurred by expulsion through the cervix, since the cervix was patent on the day of loss in these mares and in the mares ovariectomized or treated with PGF(2alpha) on Day 12. In the remaining mares treated on Day 30, the intact embryonic vesicle was dislodged on Day 31 or 32. The dislodged vesicle was mobile within the uterus and was frequently found in the uterine body. The fluid volume of the dislodged vesicle gradually decreased, and the fluid was no longer detected by Day 38 to 42. Some of the placental fluids may have been eliminated by resorption since the cervix remained closed while the fluid volume decreased.     

Sonographic fetometry in the bovine

The fetuses of 19 heifers were examined a total of 485 times during the course of pregnancy by means of transrectal sonography with sector scanners. Changes in size of organs and parts of the body were fetometrically assessed and the relative frequency of different intrauterine positions during pregnancy was registered. Until the end of Month 4 of gestation, anterior and posterior presentations occurred with equal frequency. During Months 5 to 7 the posterior presentation was found in about 25% of the cases, while after that it was rarely encountered at all. The head, thorax, abdomen and pelvis of the fetuses were readily accessible for transrectal sonography during the first 4 mo. During Months 5 to 7, the thoracic, abdominal and pelvic regions could be examined in at least one third of the cases. The head was accessible in about 80% of the cases examined throughout all stages of pregnancy. Ultrasound fetometry served to assess average growth-rate of a number of organs during pregnancy. The regressions and correlation coefficients between the development of bovine fetuses and the age of gestation were obtained for the following structures: eye, braincase, trachea, stomach, vena cava, urinary bladder, scrotum, ribs, cervical vertebrae, thoracic vertebrae, lumbar vertebrae, coccygeal vertebrae, scapula, humerus, radius and ulna, metacarpus, os ilium, os ischii, femur, tibia, metatarsus, trunk, crown-rump length and umbilical cord. Additionally, the development of the fetal heart rate during pregnancy was observed. It can be concluded that intrauterine development of the bovine fetus and its gestational age may be judged from the size of its organs and parts of the body.     

Deslorelin on Day 8 or 12 postovulation does not luteinize follicles during an artificially maintained diestrous phase in the mare

Practical estrus synchronization schemes are needed for mares. The Ovsynch synchronization protocol for cattle involves the administration of gonadotropin-releasing hormone (GnRH) to induce ovulation or luteinization of dominant follicles during the luteal phase and prostaglandin 7 days later to cause regression of any luteal tissue and development of a preovulatory follicle. An Ovsynch-type synchronization program potentially could be developed for horses if luteinization or ovulation of diestrous follicles occurred in response to GnRH treatment. The objective of this study was to determine if administration of the GnRH agonist, deslorelin acetate, on Day 8 or 12 postovulation would induce luteinization or ovulation of diestrous follicles in the mare. The model used was cycling mares maintained in an artificial luteal phase by administration of a synthetic progestin following prostaglandin-induced luteal regression. On the day of ovulation, 21 light horse mares were randomly assigned to one of three groups: (1) no GnRH, altrenogest from Days 5 to 15 postovulation with prostaglandin on Day 15; (2) GnRH on Day 8, altrenogest from Days 5 to 15 with prostaglandin given on Day 6 to induce luteolysis of the primary corpus luteum, an implant containing 2.1mg of deslorelin acetate inserted on Day 8 and removed on Day 10, with a second prostaglandin treatment on Day 15; (3) GnRH on Day 12, altrenogest from Days 9 to 19, prostaglandin on Day 10, a deslorelin acetate implant injected on Day 12 (subsequently removed on Day 14), and a second dose of prostaglandin administered on Day 19. Follicular development was monitored every other day from Day 5 until a 30-mm sized follicle was observed, and then daily to detection of ovulation. Serum progesterone concentrations were determined daily for 12 consecutive days. Progesterone concentrations in Group 1 remained elevated until approximately Day 12 postovulation. Prostaglandin administration on Day 15 resulted in complete luteolysis in all seven mares. In Group 2, progesterone concentrations in six of seven mares declined to baseline after prostaglandin treatment. No increase in serum progesterone was noted in any of the six mares that were given GnRH on Day 8, including three mares that had diestrous follicles > or =30mm in diameter at the time of treatment. Similarly, progesterone concentrations in six of seven mares in Group 3 declined to baseline after prostaglandin and there was no increase in progesterone after administration of GnRH on Day 12. No ultrasound evidence of luteinization or ovulation of diestrous follicles were noted after GnRH administration in any mares of Group 2 or 3. In conclusion, administration of the GnRH agonist deslorelin acetate to mares failed to induce luteinization or ovulation of diestrous follicles. Consequently, the Ovsynch program (as used in cattle) has little efficacy for synchronization of estrus in mares.     

Initiation of superovulation in mares 5 or 12 days after ovulation using equine pituitary extract with or without GnRH analogue

Cyclic mares were assigned to 1 of 3 treatments (n=15 per group): Group 1 received equine pituitary extract (EPE; 25 mg, i.m.) on Day 5 after ovulation; Group 2 received EPE on Day 12 after ovulation; while Group 3 received 3.3 mg of GnRH analogue (buserelin implant) on the day of ovulation and 25 mg, i.m. EPE on Day 12. Mares in each group were given 10 mg PGF(2)alpha on the first and second day of EPE treatment. The EPE treatment was continued daily until the first spontaneous ovulation, at which time 3,300 IU of human chorionic gonadotropin (hCG) were given to induce further ovulations. Mares in estrus with a >/=35 mm follicle were inseminated every other day with pooled semen from 2 stallions. Embryo recovery was attempted 7 days after the last ovulation. Follicular changes and embryo recovery during 15 estrous cycles prior to treatment were used as control data. During treatment, the number of follicles >/=25 mm was higher (P<0.05) for Day 5 than for Day 12 or control mares, but the number for Day-5 mares was similar (P>0.05) to that of mares treated with buserelin implants (Group 3). Initiation of EPE treatment on Day 5 resulted in a greater (P<0.05) number of ovulation (2.9) than on Day 12 (1.1) or in the control mares (1.3) but not in the buserelin-treated mares (1.8). The number of embryos recovered from mares in the Day 5 (1.2), Day 12 (1.0), buserelin (0.9) and control (0.9) groups was similar (P>0.05). The conclusions were 1) EPE initiated in early diestrus increased follicular development and ovulation and 2) treatment with GnRH analogue marginally improved response to EPE treatment.     

Peripheral changes in estrone sulfate concentration during the first trimester of gestation in cattle: comparison with unconjugated estrogens and relationship to fetal number

Estrone sulfate originates mainly in the conceptus during gestation in cattle. Its concentration in maternal body fluids is a useful indicator of placental function. The objective of this study was to determine the profiles of estrone sulfate during early gestation in singleton and twin bearing cows using a newly developed extraction method. One or two blastocysts produced in vitro were nonsurgically transferred to regularly cycling Holstein cows on Day 7 (Day 0 was defined as the first day of standing estrus). Pregnancy was diagnosed on Days 30, 45 and 60 by transrectal ultrasonography and finally confirmed at parturition. Six cows with singleton and six with twin pregnancies were used in the experiment. Blood was collected every other morning by jugular venipuncture from the day after transfer to Day 100. Harvested plasma was applied to reversed-phase C18 cartridges. Estrone sulfate and unconjugated estrogens (estrone and estradiol-17beta) retained in the cartridge were eluted separately by methanol stepwise gradient and each measured by validated radioimmunoassay. On average, estrone sulfate concentrations fluctuated between 2 and 6 pg/ml until Day 50 in both groups and then gradually increased. However, the levels of estrone and estradiol- 17beta remained low (1-5 pg/ml) until Day 80. The concentration of estrone sulfate after Day 50 was significantly affected by the day of gestation (P < 0.0001) and the number of fetuses (P < 0.01). After Day 80. estrone sulfate increased drastically, followed by increases in estrone and estradiol-17beta concentrations. The rate of increase in estrone sulfate during Days 80-100 was the greatest among all estrogens (P < 0.05). The rates of increase in estrone sulfate during Days 50-80 and 80-100 were 1.7 times greater in twin pregnancies than in cows having one fetus. These results suggest that the concentration of estrone sulfate in bovine peripheral blood plasma during early gestation has potential application in monitoring embryonic growth as well as fetoplacental development.     

Follicular and FSH responses to parturition during the anovulatory season in mares

The ovaries of periparturient pony mares (n=9 to 16 parturitions per month for January to April) were scanned ultrasonically on the day of parturition, while those of postpartum and control mares (n=12) were examined at least twice weekly. Four mares had apparent lactational anovulation (incidence, 7%) that corrected spontaneously (1 mare) or within 14 d after the weaning of foals on August 10 (3 mares). All but 2 of the postpartum ovulations occurred after April 29; that is, parturition did not effectively stimulate ovulation in ponies foaling during the anovulatory season. Mean diameter of the largest follicle per month increased (P<0.001) progressively in the controls (means: 11.4, 14.4, 19.0 and 24.5 mm for January to April, respectively). In the parturient mares, the diameter of the largest follicle on day of parturition did not increase over months (range of means: 13.6 to 16.9 mm), indicating that a suppressive effect of pregnancy counteracted the stimulatory effect of season. Within each month of parturition, diameter of the largest follicle increased (P<0.05) between Day 0 (day of parturition) and Day 3 or 7. Blood samples for FSH assay were taken daily for 14 d from 6 mares with parturition in the middle of each month and from 6 controls on the corresponding calendar days. In periparturient mares, a significant increase in mean FSH concentrations occurred for all months of parturition between Day-2 and Day 0, followed by a significant decrease between Day 3 and Day 7. Maximum means for the periparturient FSH profile were temporally related to the beginning of follicular growth. In the controls, FSH concentrations were not affected by month or day, or by their interaction. Within each month, mean FSH concentrations were lower (P<0.05) in the periparturient mares than in the controls (averaged over all months: 3.9 +/- 0.1 versus 7.9+/-0.3 ng/ml) even though follicular growth was greater following parturition than during the corresponding calendar days in controls.     

Effects of day of estrous cycle, time of day, luteolysis, and embryo on uterine contractility in mares

One-minute continuous ultrasonic scans of longitudinal sections of the uterine body were videotaped, and contractility scores (1 to 5, minimal to maximal contractility) were assigned without knowledge of mare identity, day of the estrous cycle or pregnancy status. Contractility was assessed, and plasma progesterone concentrations were determined for each of 3 daily examinations (at 0800, 1600 and 2400 hours) from Day 9 to Day 19 (Day 0 = day of ovulation). For both the nonbred (n=11) and pregnant (n=11) mares, there was no effect of hour of scan on the extent of uterine contractility. When data for the nonbred mares were normalized to the onset of luteolysis (defined for each mare as the first >/=25% decrease in plasma progesterone concentrations between successive samples), there was an abrupt increase (P<0.05) in contractility 24 hours prior to the onset of luteolysis. Contractility was also assessed daily in 20 nonbred and 27 pregnant mares from Day 0 to Day 17. For the nonbred mares, a biphasic profile in contractility occurred during the estrous cycle as indicated by the following significant changes: a decrease between Days 0 and 2, an increase between Days 2 and 4, a plateau between Days 4 and 7, a decrease between Days 7 and 11, an increase between Days 11 and 13, and a decrease between Days 14 and 16. For pregnant mares, contractility increased (P<0.05) prior to the late-diestrous increase for nonbred mares. In addition, a significant reduction in contractility was detected on Day 5 in these mares compared with that in the nonbred mares. Contractility in the uterine body in 7 mares was assessed every 5 minutes after departure of the embryonic vesicle from the uterine body. Levels of contractility in the uterine body were lower (P<0.05) 55 minutes after the vesicle had exited the body than 相似文献   

Folliculogenesis during the transitional period and early ovulatory season in mares

Individual follicles were monitored by ultrasonography in 15 mares during the transitional period preceding the first ovulation of the year and in 9 mares during the first interovulatory interval. During the transitional period, 7 mares developed 1-3 anovulatory follicular waves characterized by a dominant follicle (maximum diameter greater than or equal to 38 mm) that had growing, static, and regressing phases. The emergence of a subsequent wave (anovulatory or ovulatory) did not occur until the dominant follicle of the previous wave was in the static phase. After the emergence of the subsequent wave, the previous dominant follicle regressed. The mean (+/- s.d.) length of the interval between successive waves was 10.8 +/- 2.2 days. Before the emergence of waves (identified by a dominant follicle), follicular activity seemed erratic and follicles did not reach greater than 35 mm. During the interovulatory interval, 6 mares developed 2 waves (an anovulatory wave and a subsequent ovulatory wave) and 3 mares developed only 1 detected wave (the ovulatory wave). The ovulatory follicle at the end of the transitional period reached 20 mm earlier (Day - 15), grew slower (2.6 +/- 0.1 mm/day; mean +/- s.e.m.) but reached a larger diameter on Day - 1 (50.5 +/- 1.1 mm) than for the ovulatory follicle at the end of the interovulatory interval (Day - 10, 3.6 +/- 0.2 mm/day, 44.4 +/- 1.0 mm, respectively; P less than 0.05 for each end point). The interval from cessation of growth of the largest subordinate follicle to the occurrence of ovulation was longer (P less than 0.05) for end of the transitional period (9.5 +/- 0.7 days) than for the end of the interovulatory interval (6.8 +/- 0.6 days). Results demonstrated the occurrence of rhythmic follicular waves during some transitional periods and the occurrence of 2 waves during some of the first oestrous cycles of the year.     

Induction of twinning in Holstein and Japanese black cows by ipsilateral frozen embryo transfer

Induction of twinning by ipsilateral nonsurgical transfer of two frozen-thawed Japanese Black bovine embryos to each of 20 Holstein and 26 Japanese Black cows, that had been kept under a stable in private farm conditions, was examined. The cows were monitored every 20 days from Day 25 to Day 65 of gestation for pregnancy and fetus survival (estrus is Day 0). Seventy-five per cent (15 of 20), 65.0% (13 of 20) or 60.0% (12 of 20) and 61.5% (16 of 26), 53.8% (14 of 26) or 50.0% (13 of 26) of Japanese Black cows were diagnosed pregnant at 25, 45 and 65 days after transfer by ultrasonic echography. Embryonic losses were observed between Days 25 and 65 in 29.2% (7) Holstein and 31.8% (7) Japanese Black cows. The twin pregnancy rate in Holstein and Japanese Black cows decreased with time; 60.0% (9 of 15) vs. 37.5% (6 of 16) at Day 25; 53.8% (7 of 13) vs. 28.6% (4 of 14) at Day 45 and 41.7% (5 of 12) vs. 15.4% (2 of 13) at Day 65. At calving, Holstein cows produced five sets of twins and seven single calves, and Japanese Black cows two sets of twins and 11 single calves. The twinning rate in Holstein cows was higher (P < 0.05) than that in Japanese Black cows, 41.7% (5 of 12) vs. 15.4% (2 of 13). The calf birth weight in Holsteins was heavier (P < 0.05) than that in Japanese Black dams (24.5 kg, 33.6 kg vs. 19.3 kg, 25.5 kg for twin and single calves). The placental weight in Holstein dams calving twins was heavier than that in Holstein dams calving a single calf or in Japanese Black dams calving either twins or a single calf (6.6 kg vs. 3.5 kg, 4.6 kg or 2.8 kg). The number of placentome in Holstein dams calving twins was also higher (P < 0.05) than that in Holstein dams calving a single calf or Japanese Black dams (103.5 vs. 41.8, 67.9, 33.0). The number of placentome was approximately double in dams calving twins than that of dams calving a single calf.