Induction of parturition, progesterone secretion, and delivery of placenta in beef heifers given relaxin with cloprostenol or dexamethasone

Sixty primiparous beef heifers from a crossbreeding study were used to examine the effects of inducing parturition with relaxin (3,000 U/mg) combined with cloprostenol (500 micrograms, i.m., n = 30) or dexamethasone (20 mg, i.m., n = 30) at Day 273, 10 +/- 1 days before expected parturition (Day 283). Heifers were assigned at random within cloprostenol and dexamethasone groups to receive relaxin (1 mg, n = 5/treatment), i.m. or in the cervical os (OS), at 0 h (the same time as cloprostenol and dexamethasone) or 24 h later. Eleven and six first-calving heifers and sixteen and nine second-calving cows also received cloprostenol + relaxin and cloprostenol + phosphate-buffered saline, respectively. Radioimmunoassay of daily plasma samples indicated an abrupt decrease in progesterone with time (p less than 0.001), from 7.5 +/- 0.50 to 1.0 +/- 0.30 ng/ml (mean +/- SE) within 48 h for all groups. The mean rate of progesterone decrease (ng/ml in 24 h) was accelerated (p less than 0.01) in relaxin-treated heifers (5.3 +/- 0.36), in contrast to dexamethasone- and cloprostenol-treated control heifers (2.8 +/- 0.40). Relaxin combined with cloprostenol or dexamethasone shortened the calving period in these heifers by reducing the interval between treatment and calving (33 vs. 56 h; p less than 0.01). The incidence and duration of retained placenta were reduced by 22 vs. 75% and 14 vs. 34 h for relaxin combined with cloprostenol or dexamethasone as compared with cloprostenol- or dexamethasone-treated controls, respectively (p less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)     

Endocrine parameters associated with disruption of parturition after bilateral pelvic neurectomy.

Bilateral lesions of the pelvic nerve (BLPN) result in dystocia, but the processes which control this effect are not fully understood. Plasma progesterone, relaxin, and luteinizing hormone (LH) concentrations were measured in blood samples taken in the morning (AM) and evening (PM) of Days 20-23 of gestation from rats with BLPN or sham neurectomy. Ten of 11 sham-operated control animals delivered their entire litters by Day 23 of gestation, but animals with BLPN did not complete parturition by Day 23 when they were sacrificed. Progesterone concentrations were greater in rats with BLPN than in sham-operated rats on Day 20 PM and Day 21 AM, but hormone concentrations declined to minimal values by Day 22 in both groups. Relaxin concentrations were greater in rats with BLPN than in sham-operated rats on Day 21 PM. Thereafter, relaxin concentrations decreased to reach minimum values on Day 23 in both groups. LH concentrations were low throughout the period of study in rats with BLPN; however, a postpartum LH surge was detected in all sham-operated animals. Data from this study indicate that the pelvic nerve does not control parturition by modulating serum relaxin and progesterone concentrations; however, these data suggest that impulses carried by the pelvic nerve influence ovarian secretion of these hormones. In addition, these data indicate that the pelvic nerve transmits stimuli from the cervix to the hypothalamus to facilitate the postpartum LH surge.     

Pelvic growth, calf birth weight and dystocia in Holstein x Hereford heifers

The pelvic area was measured in 129 Holstein x Hereford heifers at 10, 16 and 22 months of age. The heifers were fed an all forage diet. Pelvic growth was not linear over time, changing from an increase of 0.27 +/- 0.2 cm(2)/day during the first 6 months of the study to 0.13 +/- 0.13 cm(2)/day during the last 6 months (P<0.01). The relationship of pelvic area to body weight, height at hooks, and distance from hooks to pins did not change with age, and a moderate correlation between the pelvic area and these other measures (R=0.20 to 0.80) was noted. The pelvic area was measured within 24 hours after calving in 76 of the heifers. The rate of increase of pelvic area/day increased significantly (P < 0.01) in the month prior to calving from 0.14+/-0.13 cm(2) to 1.15 +/- 0.88 cm(2). As a result, the pelvic area at calving had a moderate correlation (R=0.29 to 0.52) to the pelvic area prior to calving. Logistic regression and discriminant analysis techniques were used to model the influence of the pelvic area and calf birth weight on the incidence of dystocia. Ratio of the pelvic area at calving to calf birth weight significantly (P < 0.01) influenced the incidence of dystocia. Logistic regression techniques were not superior to discriminant analysis; both correctly predicted 73% of the cases. Pelvic area measurement at any time other than calving was not associated with dystocia (P >0.05). Pelvic area and calf birth weight are important determinants of dystocia in heifers. The high degree of variation noted in pelvic growth, in particular during the month prior to calving, resulted in low correlation between pelvic area at calving and the precalving measurement. Therefore, we were not able to predict dystocia by measuring the pelvic area prior to calving.     

Rabbit relaxin: the influence of pregnancy and ovariectomy during pregnancy on the plasma profile.

A porcine relaxin radioimmunoassay was developed to evaluate the profile of immunoreactive relaxin in rabbit plasma. Relaxin was nondetectable in pseudopregnant (Days 1, 4, 5-8, 12, and 16), nonpregnant, and male rabbits. However, in pregnant rabbits, relaxin was detected during the peri-implantation period (Days 4-9). Peak concentrations were reached on Day 15 and were maintained until parturition (Day 32). Relaxin concentrations abruptly decreased on Day 1 postpartum to low but detectable concentrations that were unchanged during the first week postpartum. In contrast, progesterone concentrations peaked earlier (Day 13), decreased after Day 25, and were not detectable on Day 1 postpartum. The effect of ovariectomy on the profile of plasma relaxin was evaluated. Four pregnant rabbits were ovariectomized (Day 13) and treated with medroxyprogesterone acetate to maintain pregnancy. As in normal pregnant rabbits, relaxin was observed initially during the peri-implantation period (Days 4-9) and increased to peak concentrations by Day 16. These concentrations were maintained until parturition and abruptly decreased on Day 1 postpartum to low yet detectable concentrations during the first week postpartum. The concentrations of relaxin in the plasma of ovariectomized medroxyprogesterone-treated rabbits were not different from those in three sham controls. These results indicate that the ovary is not a significant source of relaxin in pregnant rabbits. The unique observation of the presence of relaxin during the peri-implantation period suggests that this hormone has a role in preparing the rabbit uterus for implantation. The continued presence of relaxin during the first week postpartum may represent residual hormone, or it may suggest a physiological role during the early postpartum period.     

The dominant follicle exerts an interovarian inhibition on FSH-induced follicular development

An experiment was conducted to evaluate the role of the dominant follicle (DF) of the first wave in regulating follicular and ovulatory responses and embryonic yield to a superovulation regime with FSH-P. Twenty normally cycling Holstein-Freisian heifers (n = 20) were synchronized with GnRH and pgf(2alpha) and randomly assigned to a control or a treated group (n = 10 each). Treated heifers had the first wave dominant follicle removed via transvaginal, ultrasound-guided aspiration on Day 6 after a synchronized estrus. All heifers received a total of 32 mg FSH-P given in decreasing doses at 12 h intervals from Day 8 to Day 11 plus two injections of pgf(2alpha) (35 mg and 20 mg, respectively) on Day 10. Heifers were inseminated at 6 h and 16 h after onset of estrus. Follicular dynamics were examined daily by transrectal ultrasonography from Day 4 to estrus, once following ovulation, and at the time of embryo collection on Day 7. Blood samples were collected daily during the superovulatory treatment and at embryo collection. Follicles were classified as: small, /= 10 mm. Aspiration of the dominant follicle was associated with an immediate decrease in large follicles, and a linear rate increase in small follicles from Day 4 to Day 8 just prior to the FSH-P injections, (treatment > control: +0.33 vs. -0.22, number of small follicles per day; P < 0.10). During FSH-P injections, the increase in number of medium follicles was greater (P < 0.01) for treatment on Day 9-11 (treatment > control: Day 9, 3.2 > 1.8; Day 10, 9.2 > 4.7; Day 11, 13.1 > 8.3; +/- 0.56). Number of large follicles was greater in treatment at Day 11 (5.12 > 1.4 +/-0.21; P < 0.01). Mean number of induced ovulatory follicles (difference between number of follicles at estrus and Day 2 after estrus) was greater in treatment (13.4 > 6.3 +/- 1.82; P < 0.01). Plasma estradiol at Day 11 during FSH-P treatment was greater in treatment (32.5 > 15.8 +/- 2.6; P < 0.01). Plasma progesterone at embryo flushing (Day 7 after ovulation) was greater in treatment (7.4 > 4.9; P < 0.02); technical difficulties at embryo recovery reduced sensitivity of embryonic measurements. No changes in the distribution of unfertilized oocytes and embryo developmental stages were detected between control and treatment groups. Presence of dominant follicle of the first wave inhibited intraovarian follicular responses to exogenous FSH.     

Ultrasonic morphology of corpora lutea and central luteal cavities during the estrous cycle and early pregnancy in heifers

Ultrasonography was used once daily to quantify corpora lutea, central luteal cavities, and luteinized tissue during interovulatory intervals (n = 66) and during Days 0 to 60 of pregnancy (n = 14) in nulliparous Holstein heifers (ovulation = Day 0). The corpus luteum of the estrous cycle was detectable by ultrasonography in most heifers from the day of ovulation (mean, Day 0.5) and extending into the regressive phase beyond the next ovulation (mean, Day 1.4 +/-0.2 after the next ovulation). During pregnancy, the corpus luteum was detected until Day 60 (end of study). Maximal central luteal cavity area detected on Days 0 to 20 was used retrospectively to group luteal glands into four cavity categories: no, small, medium, and large. These categories corresponded to approximate cavity diameters of <2 mm, 2 to 5 mm, 6 to 10 mm, and >10 mm, respectively. The incidence of each cavity category was similar between interovulatory intervals and pregnancies (combined incidence, 17 80 , 8 80 , 33 80 , and 22 80 for no, small, medium, and large cavities, respectively; total with cavities, 63 80 , 79%). Mean day of first detection of a central cavity was earliest for large cavities during interovulatory intervals (means, Days 4.7, 4.4, and 3.0 for small, medium, and large cavities, respectively; P<0.04) and during pregnancies (means, Days 5.5, 4.2, and 3.3, respectively; NS). However, the day that the cavities reached maximum size (range of means, Days 5.5 to 7.0) did not differ among categories. Mean day of last detection of the central cavity was significantly different among cavity categories during interovulatory intervals (means, Days 9.3, 11.1, and 17.4 for small, medium, and large cavities, respectively) and pregnancies (means, Days 7.0, 8.8, and 20.2, respectively). Time of loss of central cavities was similar between nonbred and pregnant heifers, and there was no significant difference among cavity categories in the length of the interovulatory interval (mean, 20.1 d). Luteal tissue area was not significantly different among cavity categories during interovulatory intervals. There were no indications that cavities were functionally important. Luteal tissue area increased linearly in pregnant heifers on Days 21 to 60 (mean slope, 2.6 mm(2)/day).     

First-service pregnancy rate in beef heifers as influenced by human chorionic gonadotropin administration before and/or after breeding

Two experiments were conducted to evaluate whether administration of human chorionic gonadotropin (hCG) before and/or after breeding influences the first-service pregnancy rate in beef heifers. In Experiment 1, 125 yearling and two-year-old heifers were allotted to one of four groups: a control group; a group receiving 3,000 IU hCG on Day 4 of the prebreeding estrous cycle; a group receiving 3,000 IU hCG on Day 4 post breeding; and a group receiving 3,000 IU hCG on Day 4 of the prebreeding estrous cycle and again on Day 4 post breeding (Day 1 = estrus). First-service pregnancy rate was not affected by a single intramuscular (i.m.) injection of 3,000 IU of hCG on Day 4 of the prebreeding estrous cycle and/or post breeding. In Experiment 2, 111 yearling heifers were allotted either to an untreated control group or to a group receiving 3,000 IU hCG on Day 4 post breeding. Administration of a single i.m. injection of hCG on Day 4 post breeding did not affect the first-service pregnancy rate.     

Relaxin regulates oxytocin secretion in late-pregnant beef heifers

The effects of porcine relaxin (3000 units/mg) on oxytocin (OT) and progesterone secretion were studied in beef heifers on Day 274 (10 days before expected parturition). Heifers (n = 11) were randomly assigned to three treatments: relaxin iv infusions combined with im injection (RLX-INF, 9000 units), relaxin im injection (RLX-im, 6000 units), and phosphate-buffered saline-treated controls (PBS). RLX-INF heifers received infusions of PBS and 1000 units of relaxin for 165 min, followed by 2000 units of relaxin im and finally 2000 units of relaxin infusion followed by 4000 units of relaxin im. Endogenous relaxin (immunoreactive) in the PBS-treated group was 0.2-0.9 ng/ml peripheral plasma. For the RLX-im group, peak relaxin was 81 +/- 12 ng/ml (+/- SE) at 45 min after treatment. There were two peaks of relaxin, 18 +/- 5.3 ng/ml and 74 +/- 7.5 ng/ml, 3.5-4.5 hr apart in the RLX-INF group. Significant peak releases of OT were evident in the relaxin-treated heifers. For the RLX-im group, an OT peak (42 +/- 16 pg/ml) occurred within 30 min after relaxin treatment. For the RLX-INF heifers, 2000 and 4000 units of relaxin were associated with major peaks of 14 +/- 0.5 and 43 +/- 1.7 pg/ml OT, respectively. Basal OT plasma levels in the PBS group were 2.5-3.1 pg/ml. Mean plasma progesterone for all heifers was 6.2 +/- 2.11 ng/ml before treatment. There was a significant decrease in progesterone (-2.5 ng/ml) in the RLX-im group within 60 min after relaxin treatment and 45 min after peak OT secretion. The maximum decrease in progesterone (-3.2 +/- 0.68 ng/ml) occurred 135 min after treatment in the RLX-im group. In the RLX-INF group, 2000 units of relaxin infusion combined with 4000 units of relaxin im significantly decreased progesterone (-3.2 +/- 1.59 ng/ml) in peripheral plasma. These results clearly indicate that relaxin causes an acute peak release of oxytocin within 30 min, followed by a marked decrease in plasma progesterone concentration in late-pregnancy cattle.     

Factors affecting superovulation in heifers treated with PMSG

In this study we determined 1) if the immunoneutralization of PMSG affected the ovulatory response, the number of large follicles and embryo yield compared with that of PMSG alone or pFSH, and 2) whether the stage of the estrous cycle at which PMSG was injected affected the ovulatory response and yield of embryos in superovulated heifers. Estrus was synchronized in 99 (Experiment 1) and 71 (Experiment 2) heifers using prostaglandin F2alpha (PG) analogue, cloprostenol, given 11 d apart in replicate experiments over 2 yr. In Experiments 1 and 2, heifers were randomly allocated to 1 of 3 treatments (initiated at mid-cycle): Treatment 1--24 mg of pFSH (Folltropin) given twice daily for 4 d; Treatment 2--a single injection of 2000 IU PMSG; Treatment 3--2000 IU PMSG followed by 2000 IU of Neutra-PMSG at the time of first insemination. In Experiment 3, 116 heifers were given 2000 IU PMSG on Day 2 (n = 28), Day 3 (n = 27), Day 10 (n = 41) or Day 16 (n = 20) of the estrous cycle. The PG was given at 48 h (500 microg cloprostenol) and 60 h (250 microg cloprostenol) after the first gonadotropin treatment. Heifers were inseminated twice during estrus, and embryos were recovered on Day 7, following slaughter and graded for quality. The numbers of ovulations and large follicles (> or =10 mm) were also counted. There was no effect of treatment on ovulation rate in Experiment 1, but in Experiment 2 it was greater (P < 0.002) in heifers given PMSG (14.7 +/- 1.5) than pFSH (7.5 +/- 1.4) or PMSG-neutra-PMSG (8.7 +/- 1.5). The number of large follicles was higher following PMSG than pFSH treatment in Experiment 1, and it was higher (P < 0.004) in heifers given PMSG (5.5 +/- 0.8) than pFSH (1.12 +/- 0.7) or PMSG-neutra-PMSG (2.7 +/- 0.8) in Experiment 2. The use of Neutra-PMSG did not affect the numbers of embryos recovered or numbers of Grade 1 or 2 embryos, but it did decrease the number of Grade 3 embryos in both experiments. In Experiment 3, the ovulation rate decreased (P < 0.004) when PMSG was given on Day 3 (5.7 +/- 1.46) of the cycle rather than on Day 2 (12.3 +/- 1.64), Day 10 (13.4 +/- 1.45) or Day 16 (12.5 +/- 1.87). There was no effect of day of treatment on the numbers of large follicles. The mean numbers of embryos recovered were lower (P < 0.01) in heifers treated on Day 3 (2.1 +/- 0.67) than on Day 2 (6.8 +/- 1.0), Day 10 (6.4 +/- 0.86) or Day 16 (7.8 +/- 1.87). It is concluded that Neutra-PMSG given to heifers treated with PMSG did not improve embryo yield or quality and that treatment with PMSG early in the cycle can result in acceptable embryo yields provided sufficient time elapses between treatment and luteolysis.     

Measurement of plasma and tissue relaxin concentrations in the pregnant hamster and fetus using a homologous radioimmunoassay

A homologous hamster relaxin RIA was developed to evaluate plasma and tissue concentrations of relaxin in the latter half of pregnancy in this species. Relaxin protein and mRNA were localized using antibodies developed to synthetic hamster relaxin and gene-specific molecular probes, respectively. Molecular weight and isoelectric point of the synthetic and native hormones were identical by electrophoretic methods, and synthetic hamster relaxin was active in the mouse interpubic ligament bioassay. Synthetic hormone was used as tracer and standard with rabbit antiserum to the synthetic hormone in the RIA. Relaxin was assayed in blood samples recovered from the retro-orbital plexus on Days 6, 8, 10, 12, 14, 15, and 16 of gestation and on Days 1 and 5 postpartum. Relaxin was first detected on Day 8 of gestation (3.7 +/- 0.6 ng/ml), increased to reach a maximum in the evening of Day 15 (826.0 +/- 124.0 ng/ml), and decreased by Day 16 (day of parturition). Relaxin concentrations were assayed in aqueous extracts of implantation sites (Days 6, 8, and 10) and chorioallantoic placentae (Days 12, 14, and 15). Concentrations were low on Day 6 (0.02 +/- 0.001 microg/g tissue), increased to Day 15 (6.96 +/- 0.86 microg/g tissue), and subsequently declined by the evening of Day 15. Relaxin protein and mRNA were localized to primary and secondary giant trophoblast cells in the chorioallantoic placental trophospongium. However, relaxin protein was not localized in ovaries of pregnant animals or oviductal tissues of cycling animals. Significant quantities of relaxin were detected in the serum of fetal hamsters recovered on Day 15.     

Pregnant mouse corpora lutea: immunocytochemical localization of relaxin and ultrastructure

Relaxin was localized in corpora lutea of pregnant mouse ovaries by using the unlabeled antibody peroxidase-antiperoxidase technique and a highly specific rabbit antirat relaxin serum. Relaxin immunostaining was first observed in luteal cells located at the periphery of corpora lutea on Day 10 of gestation. The number of relaxin immunostained cells and the intensity of the stain gradually increased to reach a maximum between Days 16 and 18 of gestation. While a few luteal cells were specifically stained for relaxin on Day 1 postpartum, no luteal cells were stained on Day 2 postpartum. Ultrastructural studies of luteal cells from pregnant mouse ovaries revealed the presence of a distinct electron-dense, membrane-bound granule population, which was first observed on Day 12 of gestation. The granules increased in number to reach a maximum between Days 16 and 18 of gestation, and were absent by Day 2 postpartum. The appearance and disappearance of this granule population closely paralleled the relaxin immunostaining in the luteal cells. We suggest that the granules may be the subcellular sites of relaxin storage in the pregnant mouse ovary.     

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.     

Predicting dystocia in heifers

The relationship of heifer size, calf size and sire on dystocia was studied in approximately one thousand Angus, Hereford X Angus and Charolais heifers bred to Angus, Hereford and Charolais bulls. Heifers were weighed and pelvic measurements taken 35 days post breeding, at midgestation, and on a random group of 135 heifers two weeks prior to calving. Dystocia score, calf birth weight, calf body length, calf hip width and calf sex were recorded at calving.Correlation coefficients for calf traits indicate that birth weight and birth weight to body measurement ratios were the most highly correlated variables with dystocia score. For traits of the dam, pelvic area was the most highly correlated variable to dystocia score, while dam weight had a low and nonsignificant correlation.Sires experiencing a higher than average incidence of dystocia had calves that were larger for all measures of body size. Individual sire differences for dystocia indicate a large number of calves and an accurate randomization of heifers bred to each bull is necessary to make precise judgements as to a bull's potential for siring easy-calving offspring.Calf size measurements were positively correlated with heifer weight and pelvic area; however, correlations tended to be low. Breed of calf, sire, heifer weight I and II and pelvic area I, II and III accounted for only 23.6% of the variation in calf birth weight.Charolais calves were larger (P<.05) and had more dystocia (P<.05) than Hereford or Angus sired calves. Hereford sired crossbred calves were larger (P<.05) and had more dystocia than Angus crossbred or straightbred calves. Angus crossbred calves were larger (P<.05) but had no more dystocia than Angus straightbred calves.Heritability estimates for calf body length (0.35) and calf hip width (0.42) indicate calf skeletal measurements were more heritable than calf birth weight (0.28).Pelvic area measurements were correlated (P<.01) with heifer weight; however, heifer weight I and II accounted for only 37% and 30% of the variability in pelvic area I and II, respectively. Average pelvic area was largest in Charolais heifers, followed by Crossbred and Angus heifers. Pelvic area growth was .275 cm²/day in Angus, .254 cm²/day in Crossbred, and .250 cm²/day in Charolais heifers.Dystocia score was most highly related to calf size and pelvic area I. Pooled over all breeds, a regression analysis showed 37% of the variability in dystocia score being accounted for by the ratio of calf birth weight to calf body length and pelvic erea I. Very little increase in the R² value occurred by the addition of the other independent variables.Histogram analyses demonstrates that a constant level of dystocia can be maintained only if pelvic area increases in proportion to calf size. However, regardless of the size of calf, heifers that had very small pelvic openings had a high rate of dystocia, just as heifers having very large calves experienced high rates of dystocia even when they had large pelves.     

Ultrasonographic determination of ovarian follicular. Development in superovulated heifers pretreated with FSH-P at the beginning of the estrous cycle

On Day 3 of the estrous cycle (estrus = Day 0), dairy heifers were given either 10 mg i.m. FSH-P (FSH-P primed; n = 9) or a saline vehicle (saline primed; n = 9). On Day 10, all heifers were superovulated with FSH-P (total = 27.7 mg i.m.) in declining doses over 5 d. Heifers were inseminated artificially at estrus. From Day 2 until estrus, the number and size of follicles >2 mm were monitored daily by ultrasonography. The mean (+/- SEM) number of corpora lutea (CL) (6.2 +/- 1.5 vs 10.7 +/- 0.9; P<0.05) and the mean number of recovered embryos and unfertilized ova (3.6 +/- 1.7 vs 8.4 +/- 2.2; P<0.05) were lower in FSH-P-primed than in saline-primed heifers. Prior to initiation of superovulation, follicles >10 mm appeared on Days 6 to 7 in saline-primed heifers but only on Days 8 to 10 in FSH-P-primed heifers (P<0.05). Also, until Day 10, the mean number of follicles 4 to 6 mm and 7 to 10 mm was higher (P<0.05) in FSH-P-primed than in saline-primed heifers. After initiation of the superovulatory treatment (Day 10 to estrus), saline-primed heifers had a greater and faster increase in the mean number of follicles >10 mm (P<0.02) than FSH-P-primed heifers did. Depletion in the number of follicles 2 to 3 mm (P<0.001) between Day 10 and estrus and in the number of follicles 4 to 6 mm (P<0.05) between Day 12 and estrus occurred in both groups of heifers. Decreased superovulatory response and embryo recovery in FSH-P-primed heifers may have been due to the presence of large follicles (>10 mm) prior to the initiation of the superovulatory treatment which reduced the ability of small follicles to grow into larger size classes during superovulatory treatment.     

Effect of alfaprostol on postpartum reproductive efficiency in brahman cows and heifers

Brahman cows (n = 54) and heifers (n = 18) were randomly allotted by calving date, sex of calf and age to one of four treatment groups. Group 1 received no treatment (control), Group 2 received 5 mg alfaprostol (AP) i.m. on Day 21 postpartum, Group 3 received 5 mg AP i.m. on Day 32 postpartum and Group 4 received 5 mg AP i.m. on both Days 21 and 32 postpartum. Blood samples were collected via tail vessel puncture at 30 min-intervals for 8 h from half the animals in each group on Days 21 and 32 postpartum, with AP injection administered 2 h after sampling had begun. All cows were bled at weekly intervals. Samples were processed to yield serum and stored at -20 degrees C until assayed for luteinizing hormone (LH) or progesterone (P(4)). All cattle were maintained with epididymectomized marker bulls and were artificially inseminated (A.I.) at first estrus. Serum P(4) was below 1 ng/ml prior to AP treatment in all animals and did not differ (P > 0.10) between treatments. Alfaprostol treatment affected mean postpartum interval (from parturition to return to standing estrus and subsequent corpus luteum formation with serum progesterone concentrations > 1 ng/ml; P < 0.08). The control group (84.8 +/- 7.9 d) did not differ from Group 2 (86.3 +/- 11.1 d) or Group 3 (66.7 +/- 5.5 d) but did differ (P < 0.09) from Group 4 (65.1 +/- 6.4 d). Cattle injected on Day 32 had a shorter (P < 0.01) postpartum interval than those not receiving treatment on that day (65.9 +/- 4.2 vs 85.7 +/- 6.8 d). Pregnancy rate was affected (P < 0.05) by AP treatment. The control group (72.2%) did not differ (P > 0.10) from any group but, Group 2 (50.0%) was lower (P < 0.04) than Group 3 (83.3%) and (P < 0.02) Group 4 (88.9%). Cattle treated on Day 32 (Groups 3 and 4) had a higher (P < 0.02) pregnancy rate (86.1%) than those not treated on Day 32 (Groups 1 and 2; 61.1%). Serum LH was affected by day (P < 0.0003) and treatment by day (P < 0.07) but not by time (P > 0.10). Treatment Group 3 (P < 0.08) and Group 4 (P < 0.0003) mean LH concentrations differed between Days 21 and 32 postpartum. Cattle receiving AP treatment on Day 32 postpartum had a higher (P < 0.04) cumulative frequency of return to estrus by 100 days postpartum than nontreated cattle.     

Effect of relaxin on facilitation of parturition in dairy heifers

Purified pig relaxin (3000 U/mg) was injected i.m. into pregnant Holstein dairy heifers on Day 276 or 277 to determine its effect on parturition and sequential measurements of the pelvic area, cervical dilatation, and peripheral blood-plasma concentrations of progesterone and relaxin. Treatments included phosphate-buffer saline (2 ml, Group C, N = 7), relaxin once (1 mg, Group 1R, N = 7), and twice (2 mg, 12 h apart; Group 2R, N = 7). Intervals (mean +/- s.e.) between the first injection of relaxin or PBS and calving were 64 +/- 17, 80 +/- 19 and 125 +/- 34 h for Groups 2R, 1R and C, respectively. The calving intervals were reduced in Groups 2R (P less than 0.01) and 1R (P less than 0.05) compared with Group C. The incidence of dystocia was 29% (2 of 7) in Group 2R and 43% (3 of 7) in Group 1R compared with 57% (4 of 7) in Group C (P less than 0.01). Body weights and ratios of males to females of the calves were similar (P greater than 0.05) between groups. Progesterone plasma concentrations decreased (P less than 0.01) earlier in Groups 1R and 2R compared with Group C, and this acute decrease began within 6 h of treatment. At 24 h after relaxin or PBS injection, progesterone concentrations were 2.7 +/- 1.1 ng/ml for Group 2R, 3.5 +/- 0.9 ng/ml for Group 1R, and 6.0 +/- 0.1 ng/ml for Group C. Relaxin reached peak blood-plasma levels of 19 +/- 2.2 ng/ml 1 h after injection of relaxin, but remained unchanged, 0.3 +/- 0.01 ng/ml, in Group C. Pelvic area was increased 26%, 22% and 14% and cervical dilatation was increased 109%, 76% and 53% 48 h after injection in Groups 2R, 1R and C, respectively, but these responses were similar among groups at the time of parturition. We conclude that two i.m. injections of relaxin facilitated earlier calving, acutely decreased progesterone secretion, increased cervical dilatation and pelvic area expansion, and decreased the incidence of dystocia in dairy heifers.     

Uterine involution and ovarian changes during early post partum in autumn-lambing Corriedale ewes

The time of uterine involution and the changes in ovarian follicle populations were studied during early postpartum in multiparous, suckling Corriedale ewes lambing in the autumn. On Day 1 (n=5), Day 5 (n=4), Day 17 (n=4) and Day 30 (n=3) postpartum ewes underwent surgery to obtain ovaries and uteri. The weights of uteri and the lengths of the previous pregnant and nonpregnant horns were recorded as well as the presence of ovarian follicles larger than 1 mm in diameter. Uterine weight and length of uterine horns decreased (P2 to < 4 mm) follicles were also present. At days 17 and 30, aside from the small and medium follicles, all the ewes also had large (>/= 4 mm) follicles, and, at Day 30 2 ewes had large corpora lutea. We conclude that in autumn-lambing Corriedale ewes macroscopic uterine involution was complete around Day 17 post partum and that follicle development begins immediately after parturition, reaching preovulatory size before Day 17. In 2 of the 3 ewes studied until Day 30, ovulation (first progesterone increase) occurred after Day 17 (Days 18 and 25).     

Effect of early luteolysis in progesterone-based timed AI protocols in Bos indicus, Bos indicus x Bos taurus, and Bos taurus heifers

The objective of this study was to evaluate the effects of treatment with an intravaginal progesterone-releasing device (CIDR) and estradiol benzoate (EB) on follicular dynamics in Bos indicus (n=23), Bos taurus (n=25), and cross-bred (n=23) heifers. To assess the influence of reduced serum progesterone concentrations during 8 days of treatment with a progesterone-releasing device on follicular dynamics, half of the heifers received PGF at CIDR insertion (Day 0; 3 x 2 factorial design). Mean (+/-S.E.M.) serum progesterone concentrations during CIDR treatment varied (P<0.05) among genetic groups: B. indicus (5.4+/-0.1 ng/mL), B. taurus (3.3+/-0.0 ng/mL), and cross-bred (4.3+/-0.1 ng/mL). Maximum diameter of the dominant follicle (DF) was smaller (P<0.01) in B. indicus heifers (9.5+/-0.5 mm) than in cross-bred (12.3+/-0.4 mm) or B. taurus heifers (11.6+/-0.5 mm). B. indicus experienced lower (P<0.01) ovulation rate (39.1%) than did B. taurus (72.7%) and cross-bred (84.0%). Heifers treated with PGF on Day 0 had lower (P<0.05) serum progesterone concentrations during progesterone treatment. The PGF treatment on Day 0 increased (P<0.01) the diameter of the DF (11.9+/-0.4 mm vs. 10.5+/-0.4 mm). Moreover, greater (P=0.02) ovulation rates (78.8 vs. 54.0%) occurred in heifers treated with PGF on Day 0. In summary, B. indicus heifers had greater serum progesterone concentrations, smaller DF diameter, and a lower ovulation rate compared to B. taurus heifers. Prostaglandin treatment on the day of CIDR insertion reduced serum progesterone during treatment, and resulted in increased maximum DF diameter and ovulation rate.     

Prostaglandin f(2alpha) concentrations, fatty acid profiles, and fertility in lipid-infused postpartum beef heifers.

Effects of lipid infusion into postpartum (PP) beef heifers on plasma concentrations of linoleic acid and prostaglandin (PG) F(2alpha) metabolite (PGFM), days to first estrus, and subsequent pregnancy rate were examined. Treatments (n = 5 per group) of 1 L intralipid (20% soybean oil; IL), 1 L 50% dextrose (DEXT; isocaloric to IL), 0.5 L intralipid (0.5 IL), and 1 L physiological saline (SAL) were infused i.v. over 4 h on each of Days 7 through 11 PP. Capacity of the uterus to produce PG was evaluated after i.v. injection of 150 IU of oxytocin (OT) to IL- and DEXT-treated heifers Day 12 PP. Change in plasma concentrations of PGFM from 0 to 4 h was greater for IL-treated heifers than for heifers given other treatments on Day 7 (P = 0.04) and on Day 11 (P = 0.01), but not on Day 9 (P>0.10). Plasma linoleic acid on Day 11 and OT-induced release of PGFM on Day 12 were greater in IL-treated heifers compared with DEXT-treated heifers (P<0.06 and P = 0.01, respectively). There were no significant differences among treatments for mean days to first estrus or pregnancy rate. Infusion of lipid increased systemic concentrations of linoleic acid and increased the capacity of PP heifers to produce uterine PGF(2alpha) as indicated by plasma PGFM concentration after OT injection.     

Determination of the source of immunoreactive relaxin in the cat

We have recently reported the secretory profile of relaxin throughout gestation in the cat. Because the appearance of relaxin begins at about Day 20 (Day O = ovulation) and because implantation begins shortly before this at Days 13-14, we hypothesized that relaxin was of feto-placental origin. To test this hypothesis, we used 4 experimental groups: 1) Control (laparotomy-only at Day 23 or 42, n = 4); 2) Early Ovariectomy (Ovx, bilateral ovariectomy between Days 23 and 26, n = 4); 3) Late Ovx (bilateral ovariectomy between Days 40 and 44, n = 4); 4) Tissue Removal (removal of feto-placental units, uterus, and one ovary on Days 16, 21, 28 and 35, n = 1 per day). Pregnancies were maintained in both Ovx groups by progesterone administration. Relaxin secretory patterns in Ovx groups were similar to the Control data. Relaxin was detectable in plasma beginning at about Day 20, with maximum concentrations reached by Day 30. Relaxin concentrations were highest (immunoactivity per mg tissue) in homogenates of placental tissues as compared to luteal, fetal, or uterine tissues. Altogether, these data indicate that the feto-placental unit is the source of relaxin in the cat.