Side of ovulation at each end of two- and three-wave interovulatory intervals and before and after pregnancy in cattle.

The side of ovulation (left ovary, LO; right ovary, RO) and side of the next ovulation were compared between (1) beginning and end of an interovulatory interval (IOI) and beginning and end of consecutive sets of two and three IOI (n = 900 IOI), (2) beginning and end of the IOI for two and three follicular waves per IOI (n = 1300), and (3) beginning of pregnancy and first postpartum ovulation (n = 793). Pairs of sides of ovulation were designated LL (LO and LO), RR, LR, and RL. The frequency of ovulation pairs for two ends of an IOI was not different from two ends of two or three consecutive IOI indicating that differences between LO and RO were more likely inherent than from factors that developed in each IOI. For each end of an IOI or two consecutive IOI, the least frequency (P < 0.05) was for LL (16 %) with no differences among RR, LR, and RL (28 % for each). Frequencies between ipsilateral (LL, RR) and contralateral (LR, RL) ovulations pairs were not different for two-wave IOI (48 % compared with 52 %) but differed (P < 0.0001) for three-wave IOI (32 % compared with 68 %) and for pregnancy/postpartum (34 % compared with 66 %). In pregnancy/postpartum, each pair was different (P < 0.05) from each other: LL (13 %), RR (21 %), LR (30 %), RL (36 %). The lesser frequency for LL than for any of the others for an IOI, consecutive IOI, and pregnancy/postpartum indicated a ubiquity of the small propensity for LO ovulation.

NR5A2 and potential regulatory miRNAs in the bovine CL during early pregnancy.

Progesterone, which is secreted from the corpus luteum, is indispensable for the establishment and maintenance of pregnancy. The orphan nuclear receptor subfamily 5 group A member 2 (NR5A2) is a regulator of murine luteinization, but neither its regulation nor its role in the fully differentiated, mature corpus luteum (CL) have been described. Therefore, the goal of this study was to profile abundance and investigate the regulation and functions of NR5A2 in the bovine CL. Treatment of cultured luteal steroidogenic cells with a pharmacological inhibitor of NR5A2 decreased progesterone production and tended to decrease abundance of HSD3B1 mRNA. Luteal NR5A2 mRNA increased and NR5A2 protein tended to increase between days 4 and 6 of the estrous cycle, coincident with increased steroidogenic capacity of the CL. Luteal NR5A2 mRNA decreased by 8 h after prostaglandin (PG) F2A injection. During early pregnancy, luteal NR5A2 mRNA was less on days 20 and 23 compared to day 14, but protein abundance did not change. Neither 1 nor 10 ng/mL interferon tau (IFNT) altered NR5A2 abundance in cultured luteal steroidogenic cells, but 10 ng/mL PGF2A decreased NR5A2. Because of discrepancies between mRNA and protein abundance of NR5A2, regulation by miRNA that changed during early pregnancy was investigated. miR-27b-3p, miR-432-5p, and miR-369-3p mimics decreased NR5A2 protein abundance and miR-369-3p also inhibited progesterone production. Overall, the results of this study show that NR5A2 may be maintained by miRNA during early pregnancy and may be an important regulator of luteal progesterone production.

Temporal changes in the corpus luteum during early pregnancy reveal regulation of pathways that enhance steroidogenesis and suppress luteolytic mechanisms†.

Although rescue of the corpus luteum (CL) is required for pregnancy, luteal function during maternal recognition of pregnancy remains largely unexplored. CL were collected from pregnant cattle on days 14, 17, 20, and 23, to encompass the maternal recognition of pregnancy period. Next-generation sequencing was used to profile mRNA abundance during this time, while tandem mass spectrometry and nanostring technology were used to profile proteins and miRNA, respectively. A total of 1157 mRNA were differentially abundant, while 27 miRNA changed, and 29 proteins tended to change. mRNA that increased were regulators of interferon signaling and DNA repair, while those that decreased were associated with luteolytic processes, such as calcium signaling and matrix metallopeptidase (MMP) signaling, indicating inhibition of these processes. One of these, MMP12, was regulated by prostaglandin F2A in vitro. mRNA that were maximally abundant on day 20 were primarily associated with immune processes. Two of these, C-C motif chemokine ligand 1 and NFKB inhibitor alpha, were regulated by interferon tau in vitro. MiRNA that increased were predicted to inhibit phosphatidylinositol signaling, while those that decreased may be negative regulators of steroidogenesis. One protein that was greater on day 20 than on day 14 was aldehyde dehydrogenase 1 family member A1 (ALDH1A1), which synthesizes retinoic acid. Pharmacological inhibition of this enzyme, or of retinoic acid receptor signaling, led to suppression of progesterone production in vitro. Overall, these data indicate that there are changes in the CL of pregnancy that are important for continued luteal function.

Effects of service sire on prenatal mortality and prolificacy in ewes.

Ability to select service sires that minimize partial or complete losses of pregnancy could have major economic impacts in sheep production systems. This study tested the null hypothesis that survival of potential progeny did not vary with breed type of service sire or among individual rams. Data included 980 ewes on 10 farms; each ewe was pregnant to 1 of 67 rams of 12 breeds. Number of conceptuses was estimated once during pregnancy by ultrasonography, either transrectal (embryos) or transabdominal (fetuses), and was compared with number of lambs born to estimate losses. Data were examined first for number of lambs born and second for documented losses. Individual service sires affected number born (P < 0.001), which varied from 0.70 to 2.45 lambs per pregnant ewe. The main effects of breed type on lambs born were not significant, but breed types of both service sires (P < 0.0002) and ewes (P < 0.001) interacted with diagnosed number of conceptuses. Lambs born varied with ewe age (P < 0.0001) and among farms (P < 0.0001), and statistically, farms interacted with number of diagnosed conceptuses (P < 0.0001); season had no effect. In documented losses, there were both main effects of individual service sire and a service sire × number of diagnosed embryos interaction (P < 0.005). Thus, ewes bred to some rams were more apt to lose single pregnancies, whereas ewes bred to other rams were more apt to lose 1 or more embryos or fetuses from multiple pregnancies. Breed type of service sire affected (P < 0.05) prenatal death. Complete losses of single conceptuses tended to be greater in ewes bred to black-faced or hair-type rams (service sire breed type × number of diagnosed conceptuses; P < 0.09). Breed type of ewes also varied in incidence of complete losses (P < 0.05); hair-type ewes (46%) lost more (P < 0.02) documented conceptuses from examination to birth than black-faced (27%), white-faced (20%), or dairy-type (25%) ewes. Greater losses of singles than of multiples occurred in black-faced (37% vs. 18%) and hair-type (64% vs. 27%) ewes than in other breeds (ewe breed type × number of conceptuses; P < 0.03) per ewe. Surprisingly, purebred conceptuses were lost less often (24%) than crossbreds (36.4%; P < 0.002). Selection of rams based on records of prenatal losses in ewes they serviced may be a method to decrease embryonic and fetal wastage. However, further study to determine repeatability of differences among service sires from year to year will be required.

Pregnancy outcome in dairy and beef cattle after artificial insemination and treatment with seminal plasma or transforming growth factor beta-1.

Reduced capability of the uterus to support pregnancy in the absence of its interaction with secretions from male accessory glands has been demonstrated in rodents and to some extent in pigs. However, in cattle, the role of postmating inflammatory response on pregnancy success has not been studied. The current study examined the influence of uterine presensitization with seminal antigens at breeding on pregnancy outcome in cows. Lactating beef (n=1090) and dairy (n=800) cows received 0.5 mL seminal plasma (SP), 40 ng recombinant human transforming growth factor-beta1 (rhTGF-beta1), or 0.5 mL bovine serum albumin (BSA), or were left untreated before or at insemination. Semen was deposited into the anterior cervix using a second insemination gun. Pregnancy was diagnosed at 35 to 40 d postinsemination by transrectal ultrasonography or from records of calves born the subsequent calving season. Pregnancy rates in beef cows did not differ among treatments but differed among trials (69.8%, 52.5% vs. 40.3%; P<0.05). In trials where average pregnancy rates were below 50%, treatments with TGF-beta1 but not SP tended (P<0.07) to increase pregnancy rates in beef cows. In dairy cows, SP and TGF-beta1 improved pregnancy outcome by 10 percentage points, but these increments did not achieve statistical significance. In conclusion, this study did not find any conclusive evidence for the effect of TGF-beta1 or seminal plasma on pregnancy outcome in lactating dairy or beef cows but realized marginal improvements when pregnancy rates were below 50% (compromised fertility).

Use of gonadotropin releasing hormone to improve reproductive responses of ewes introduced to rams during seasonal anestrus.

Three experiments were conducted on anestrous ewes of Suffolk, Dorset, and Katahdin breeding to examine the potential value of GnRH to improve ovulation and pregnancy in response to introduction of rams. In Experiment 1, treatment with GnRH 2d after treatment with progesterone (P(4); 25mg i.m.) at introduction of rams was compared to treatment with P(4) alone at the time of introduction of rams. Treatment with GnRH did not increase percentages of ewes with a corpus luteum (CL) 14d after introduction of rams, pregnant 32d after treatment with PGF(2)alpha 14d after introduction of rams, or percent of treated ewes lambing to all services. In Experiment 2, treatments with GnRH on day 2, 7, or both after introduction of rams were compared. Treatments did not differ in mean estrous response, percentages of ewes with a detectable CL or number of CL present on day 11, or mean pregnancy and lambing rates. Therefore, neither one nor two injections of GnRH at these times appeared to be effective to induce anestrous ewes to breed. In Experiment 3, treatments compared included GnRH 4d before introduction of rams, GnRH 4d before and 1d after introduction of rams, ram introduction alone, and treatment with P(4) (25mg i.m.) at the time of introduction of rams. Percentages of ewes with concentrations of P(4) greater than 1ng/mL (indicating formation of CL had occurred) 7d after ram introduction tended to be greater (P<0.07) in ewes treated with GnRH or P(4) than in control ewes treated with ram introduction alone. However, there was no difference in P(4) concentrations between groups by day 11 or 12 after introduction of rams. Estrous response rates and percentages of ewes pregnant 95d after PGF(2)alpha was administered (on day 12 after introduction of rams) tended to be greater (P=0.08 and 0.06, respectively) in ewes treated with GnRH or P(4) than in ewes exposed to rams only. There was no difference in response variables between ewes treated with GnRH 4d before introduction of rams and ewes treated with GnRH 4d before and 1d after introduction of rams. In conclusion, treatment with GnRH 4d before ram introduction showed promise as an alternative to treatment with P(4) to improve the ovulatory response and reproductive performance of ewes introduced to rams during seasonal anestrus.

Low peripheral progesterone and late embryonic/early fetal loss in suckled beef and lactating dairy cows.

Pregnancy failure during placentation in lactating dairy cows was associated with low concentrations of serum progesterone. Beef cows have greater serum progesterone and less pregnancy failure. Experiment 1 determined that reduction of serum progesterone affected late embryonic/early fetal loss in suckled beef cows. Cows (n=40) received progesterone from two new or used controlled internal drug releasing devices, replaced every 5d, beginning on Day 28 of gestation (mating=Day 0); CL were enucleated on Day 29. Retention of pregnancy was 77% in treated cows and 97% in 78 control cows (P<0.05). Experiment 2 determined how pregnant, lactating dairy cows with high or low progesterone concentrations during Days 28-34 differed in luteal function or in serum progesterone during replacement therapy. Luteal tissue from such cows was assayed for progesterone and expression of mRNA for genes of endothelin and prostaglandin (PG) systems. Secretion of progesterone and prostaglandins by dispersed luteal cells was determined during incubation with LH, endothelin-1, or arachidonic acid. Neither luteal progesterone nor mRNAs for endothelin or prostaglandin systems differed. Endothelin-1 inhibited secretion of progesterone more (P<0.05) in luteal cells from cows with low versus high serum progesterone, when incubated with arachidonic acid. Secretion of prostaglandin F(2)alpha was increased and that of 6-keto-PGF(1)alpha decreased by endothelin-1 in vitro. Serum progesterone during replacement was lower (P<0.05) for cows with low than high serum progesterone at lutectomy. Thus, clearance, more than luteal production, determined peripheral progesterone in pregnant, lactating dairy cows.

Maintenance of pregnancy in dairy cattle after treatment with human chorionic gonadotropin or gonadotropin-releasing hormone.

The objectives were to determine whether a single injection of either human chorionic gonadotropin (hCG) or GnRH would: 1) increase ancillary formation of new luteal structures, 2) increase serum concentrations of progesterone, and 3) increase pregnancy survival in dairy females treated once between 26 and 71 d of pregnancy. A total of 421 cows were enrolled between January and November 2001, with 92, 106, and 223 females (included 68 nulliparous heifers at 1 location) treated at the 3 locations. Upon diagnosis of pregnancy, females were allocated randomly to receive 100 microg of GnRH, 1,000 IU of hCG, or 2 mL of saline. Blood samples were collected at 0, 1, 2, and 4 wk after treatment, and pregnancy status was reassessed at 1, 2, and 4 wk. New luteal structures were formed in 23.8% of cattle, with hCG (50%) and GnRH (26%) being more effective than saline (7%). Treatment had no effect on the proportion of females forming 2 new luteal structures (7.6%), and 36.2% of all induced structures regressed during the 4-wk study period. Pregnancy losses were unaffected by treatment, stage of pregnancy, or number of induced luteal structures but were nearly 9-fold greater in females in which induced luteal structures regressed. No loss occurred in females having 2 new luteal structures. Pregnancy losses decreased quadratically from 30 to 42 d. Serum progesterone did not differ among treatments, but among females forming new luteal structures, progesterone was greater at 1 (7.2 +/- 0.3 vs. 6.3 +/- 0.2 ng/ mL) and 2 wk (7.0 +/- 0.3 vs. 6.1 +/- 0.2 ng/mL) after treatment. Progesterone at the first pregnancy diagnosis was predictive of imminent pregnancy loss; the lower the initial progesterone, the sooner subsequent loss was observed. The right ovary was dominant in the location of new luteal structures. Regression of new luteal structures occurred more often on the left ovary and contra-lateral to the corpus luteum of pregnancy (53.2 vs. 22%). In conclusion, treatment of dairy cattle with either GnRH or hCG failed to prevent pregnancy loss, but concentrations of progesterone were predictive of subsequent pregnancy loss.

Effects of melengestrol acetate and P.G. 600 on fertility in Rambouillet ewes outside the natural breeding season.

The effects of melengestrol acetate (MGA) and P.G. 600 on ewe fertility outside the natural breeding season were evaluated. Rambouillet ewes were assigned to one of four groups: (1) control (C; n=92); (2) PG600 (n=86); (3) MGA (n=99); and (4) MGA+PG600 (n=92). A pellet with or without MGA (0.3mg/ewe/d) was fed at 0.15kg/ewe/d for 7d. On the last day of pellet feeding, ewes were given either saline or 5mL of P.G. 600 i.m. (400IU equine chorionic gonadotropin (eCG) and 200IU human chorionic gonadotropin (hCG)). Ultrasonography was performed between Days 20 and 25 of gestation for ewes that were mated during the first 6 d of the breeding period from the MGA (n=15) and MGA+PG600 (n=8) groups, and the number of luteal structures and embryos were counted. During the first 6d of the breeding period, MGA increased (P<0.05) the percentage of ewes that mated and conceived when compared to C and PG600 (24.2% vs. 3.3% and 10.5%, respectively). Relative to MGA, the mean (+/-S.E.M.) number of luteal structures per ewe was enhanced (P<0.03) in MGA+PG600 (1.53+/-0.13 vs. 2.38+/-0.42, respectively), however as pregnancy progressed, the number of embryos (1.5+/-0.13 vs. 1.8+/-0.16, respectively) and lambs born (1.3+/-0.15 vs. 1.5+/-0.27, respectively) did not differ. Treatment with MGA reduced (P<0.01) the interval from ram introduction to lambing relative to groups that did not receive MGA (168+/-0.8d vs. 171+/-0.6d, respectively). In conclusion, treatment with MGA increased the percentage of ewes conceiving early in the breeding period. Although P.G. 600 increased the number of luteal structures present per ewe, it did not significantly enhance ewe prolificacy.

Lack of effect of transrectal ultrasonography with restraint on lambing rate and prolificacy in ewes.

The objective was to determine if transrectal ultrasonography for determination of pregnancy in restrained ewes increases embryonic/fetal death or loss of pregnancy. Ten flocks (N=873 ewes) bred in either the estrous or anestrous season were randomized, into control (C) or examined (E) groups within flock. Examined ewes were placed in a tilting squeeze chute and scanned by one of three operators for pregnancy by transrectal ultrasonography once between Days 25 and 100 post-breeding. Control ewes were not subjected to handling in the squeeze chute or pregnancy diagnosis. Two operators counted embryos in E ewes in six flocks. There were no differences between E and C ewes in percent ewes lambing or lambing rate on a per flock basis. Prolificacy per flock was greater in E ewes (P=0.05; 1.53 versus 1.60, C and E, respectively) than in C ewes. Eighty-five percent of exposed ewes lambed in the estrous season, while only 62% of ewes lambed in the anestrous season (P=0.05). Overall prolificacy did not differ with season (1.60 versus 1.53), and there was no season by treatment interaction for any variable tested. Losses averaged 0.02 per E ewe exposed in the estrous season and 0.20 in the anestrous season (P<0.05). Examined ewes in this study had similar pregnancy and lambing rates to C ewes in the same flocks and prolificacy was slightly greater. Based on these data, the combination of transrectal ultrasonography with restraint is safe for pregnancy diagnosis in ewes.

Association of fertility with numbers of antral follicles within a follicular wave during the oestrous cycle in beef cattle.

The association between conception rate at first service and numbers of follicles developed during a follicular wave was examined in 102 suckled beef cows and 14 heifers. Follicular development was monitored using ultrasonography for either two (trial 1) or three (trial 2) consecutive oestrous cycles (pre-breeding, breeding and post-breeding equivalent). Animals were examined on alternate days from day 6 after first oestrus (day 0) until ovulation and from day 6 after insemination until next ovulation or day 24 of pregnancy and were observed for oestrus twice daily and inseminated artificially at either the second (trial 1) or third oestrus (trial 2). Cows were classified as having two or three waves of follicular development for each oestrous cycle. Numbers of follicles >or=4 mm per wave were determined, and based on the maximum diameter they attained, were classified as small (4-6 mm), medium (7-10 mm) or large (>or=11 mm) follicles. Total numbers of follicles, and primarily numbers of small and medium follicles, were affected by trial and within trial by cow, oestrous cycle and follicular wave. Heifers had more small and total numbers of follicles, but fewer large follicles than cows in trial 1 (p < 0.05). The average number of antral follicles per wave in the breeding cycle or post-breeding period did not affect conception rates, which averaged 84%. Repeatability of the total numbers of antral follicles between and among oestrous cycles and follicular waves ranged from 0.01 to 0.97. In conclusion, fertility was not affected by the numbers of antral follicles >or=4 mm in diameter in a single follicular wave.

Patterns of late embryonic and fetal mortality and association with several factors in sheep.

Embryonic and fetal mortality reduce lambing rates and litter sizes, thus contributing to economic losses in the sheep industry. In the current study, the timing of late embryonic and fetal loss in ewes and the factors with which these losses were associated were examined. Ewes lambing and lambs born were compared with pregnancy diagnosis and counts of embryos by ultrasonography near d 25, 45, 65, or 85 of gestation. Approximately 19.9% of the ewes experienced late embryonic loss, fetal loss, or both; and 21.2% of the embryos or fetuses were lost from d 25 to term. Potential offspring were lost throughout gestation; 3.7% of embryos from d 25 to 45, 4.3% of fetuses from d 45 to 65, 3.3% from d 65 to 85, and 11.5% from d 85 to parturition; thus, approximately 3 to 4% of the potential offspring were lost for each 20-d period of pregnancy beyond d 25. A greater proportion of ewes lost one (36.7%) rather than all (20.5% single; 3.8% multiple) embryos or fetuses. The patterns of loss were similar in ewes mated during the anestrous season and the transitional period and did not vary with service period within breeding season or method of synchronization of estrus. Late embryonic or fetal losses were not related to the temperature-humidity index. Maternal serum collected near d 25, 45, 65, or 85 of gestation was assayed for concentrations of progesterone, estradiol-17beta , and vascular endothelial growth factor (VEGF). The proportions of embryos or fetuses lost were associated with breed type (P < 0.05), as were concentrations of progesterone (P < 0.01), estradiol (P < 0.05), and VEGF (P < 0.01). The relationships of loss or retention of pregnancy to hormonal variables at the 4 stages studied were limited. Complete and partial losses increased rapidly as maternal progesterone at d 25 decreased below 2 ng/mL (P < 0.05). Survival of fetuses within a litter from d 25 to 65 was greater for ewes with medium concentrations of VEGF near d 25 and from d 65 to parturition was greater for ewes with high concentrations of VEGF near d 45 (P < 0.05). In summary, late embryonic or fetal losses occurred from d 25 throughout gestation and varied with breed type and with concentrations of progesterone in maternal serum on d 25.

Reproductive performance of ewes after 5-day treatment with intravaginal inserts containing progesterone in combination with injection of prostaglandin f2alpha.

Three experiments were conducted with a total of 1579 ewes to examine reproductive performance in response to synchronization of oestrus during the breeding season, using controlled internal drug releasing (CIDR-G) inserts in regimens designed to provide high concentrations of circulating progesterone. In experiment 1, treatment with two CIDR-G inserts for 12 days produced conception rate (79%) and prolificacy (1.9) to first service equivalent to breeding at natural oestrus (56% and 2.0, respectively). Pregnancy rates to two service periods were 90 and 79%, respectively. In experiments 2 and 3, progesterone was delivered by a single CIDR-G insert for 5 days in combination with prostaglandin F2alpha (PGF2alpha; 5 mg i.m., twice, 3 h apart) the day before (experiment 2), or at insert removal (experiment 3). The combined treatments improved rates of synchronization of oestrus (p<0.01) by 23 and 20% points, respectively, and pregnancy rates to the first service period by 19 (p<0.05) and 13 (p<0.01) percentage points, respectively, compared to treatment with PGF2alpha alone. It is concluded that the combination of treatment for 5 days with a CIDR-G insert and two injections of 5 mg PGF2alpha, the day before, or the day of insert removal, were effective treatments to obtain high fertility at synchronized oestrus in ewes during the breeding season.

Ability of induced corpora lutea to maintain pregnancy from the third month of gestation to term in cattle.

The local relationship between the pregnant uterine horn and the CL during maternal recognition of pregnancy is well-documented. It continues beyond that time; pregnancies were maintained in lutectomized cows when CL were induced on the ovary ipsilateral, but not contralateral, to the uterine horn of pregnancy during Days 28-53. This study evaluated factors affecting maintenance of pregnancy by CL induced after Day 53, in lutectomized cows that had received exogenous progesterone from Day 29 to 15 days after induction of a CL. Twenty-four suckled beef cows were lutectomized on Day 29 of gestation; pregnancy was maintained with progesterone from two controlled internal drug releasing (CIDR) inserts, exchanged every 5 days. Beginning on Day 53, ovaries and viability of pregnancy were evaluated by ultrasonography every 5 days. When a follicle >or=10 mm in diameter was present ipsilateral to the fetus, each cow received 1,000 IU of hCG. Following induction of a CL (20 of 24), progesterone was reduced to a single CIDR for 5 days, then removed. Retention of pregnancy was confirmed by rectal palpation and calving. Cows with induced CL maintained pregnancy to term, including four with the CL contralateral to the fetus. Three cows failed to form normal CL by Day 98 and lost pregnancy after removal of exogenous progesterone. One cow that did not respond to hCG lost pregnancy during exogenous progesterone. In conclusion, CL induced after Day 53 maintained pregnancy to term, even when induced contralateral to the pregnant uterine horn.

Growth hormone at breeding modifies conceptus development and postnatal growth in sheep.

Experiments were performed to determine the effects of components of the GH-IGF axis on conceptus development and postnatal growth in sheep. In Exp. 1, ewes received one of the following treatments: 1) sustained release GH at breeding, 2) sustained release GH at breeding and estradiol-17beta at d 5 and 6, 3) only estradiol-17beta at d 5 and 6, or 4) no treatment. Uteri were flushed on d 7, and flushings were analyzed for content of IGF-I. A single injection of sustained-release bovine GH at breeding increased IGF-I content in uterine luminal flushings compared with control ewes (P < 0.05). Treatment with estradiol-17beta on d 5 and 6 after breeding did not alter IGF-I content compared with control ewes, and it blocked the effect of GH on uterine luminal IGF-I content. In Exp. 2, sustained release GH or no treatment was administered at breeding, and gravid uteri were collected at d 25, 80, or 140 of gestation. On d 80, GH-treated ewes had smaller chorioallantoic weights (P < 0.05) and tended to have more efficient placentae (fetal weight/total placental weight; P = 0.052), with a higher percentage of placental weight as cotyledons (P = 0.068) compared with control ewes. In Exp. 3, ewes were treated with or without sustained release GH at progesterone withdrawal. Lambs from GH-treated ewes were heavier at birth (P < 0.05). Lambs from GH-treated ewes reared as singles, but not lambs reared as multiples, were heavier at 30, 60 (P < 0.05), and 75 d (P = 0.075) of age than lambs from control ewes. In conclusion, ewes treated with sustained-release GH at breeding developed smaller, more efficient placentas, and had larger lambs at birth.

Preovulatory, postovulatory, and postmaternal recognition effects of concentrations of progesterone on embryonic survival in the cow.

Although fertilization rate usually is very high when male fertility is normal, pregnancy rates are below expectations when defined by the birth of live offspring in response to first service. Factors that affect establishment and retention of pregnancy include 1) preovulatory influences on the follicle and oocyte, 2) early postovulatory uterine and luteal function, 3) concentrations of hormones associated with trophoblastic and endometrial function during maternal recognition of pregnancy, and 4) less-well understood factors during the peri-attachment period. For example, decreased progesterone during preovulatory follicular development leads to a persistent follicle, premature resumption of meiosis, and a high incidence of embryonic death between the 2- and 16-cell stages. Elevated PGF(2alpha) during d 4 to 9 of the estrous cycle not only caused luteolysis but also had a direct embryotoxic effect during the morula-to-blastocyst transition. Ideal conditions during placentation and attachment are not clearly defined. Late embryonic mortality might be increased after ovulation of persistent or immature follicles. Nominal increases in secretion of PGF(2alpha) between d 30 and 35 might be important for attachment and placentation. Lower survival of embryos from wk 5 to wk 7 to 9 of gestation in the cow was associated with lower circulating concentrations of progesterone on wk 5. To maximize embryonic survival in the cow, management must provide high progesterone before estrus, quality detection of estrus, and timely insemination. Luteolytic influences of estradiol-17beta or PGF(2alpha) must be minimized early after mating and during maternal recognition of pregnancy, and high progesterone is needed during the late embryonic/early fetal period.

Effects of Gram-positive bacterial pathogens in ewes: peptidoglycan as a potential mediator of interruption of early pregnancy.

Bacterial cell walls contain peptidoglycan (PTG), which, among other actions, induces fever. The present experiment evaluated the effects of PTG treatment on early pregnancy and blood plasma concentrations of reproductive hormones. Ewes were injected i.v. with saline or 15, 30 or 60 microg kg(-1) sonicated PTG (Streptococcus pyogenes) on day 5 after mating. Each dose of PTG induced fever. Pregnancy rate at day 25 was not related to incidence of fever but tended to differ among treatments (control, 100%; low, 100%; medium, 67%; high, 60%; P < 0.08). Combined pregnancy rate in ewes from control and low dose groups (100%) was greater than that in ewes from medium and high dose groups (64%, P < 0.01). Ewes with high 13,14-dihydro-15-keto-prostaglandin F(2alpha) (PGFM) concentrations had lower pregnancy rates (6 of 10) than those with low concentrations of PGFM (11 of 11; P < 0.05). Mean cortisol concentrations were higher in treated (2.8 +/- 0.28 microg dl(-1)) than in control (1.1 +/- 0.03 microg dl(-1)) ewes (P < 0.01); the pattern of secretion was biphasic and increased in all treated ewes (P < 0.01). Neither means nor profiles of oestradiol differed with treatment. Mean concentrations and the pattern of concentrations of progesterone were reduced in all treated ewes, as indicated by the time by treatment and linear interaction with treatment (1.2 +/- 0.1 versus 1.6 +/- 0.1 ng ml(-1), P < 0.01). Patterns of LH pulses did not differ from 0 to 4 h or 24 to 28 h after treatment; mean plasma LH concentration was lower in ewes treated with 0, 15 or 30 microg PTG kg(-1) than with 60 microg PTG kg(-1) (P < 0.01). Pregnancy status was not related to plasma concentrations or patterns of LH, oestradiol, progesterone or cortisol. Inflammatory mediators, such as PGF(2alpha), may act directly on the embryo or uterus in ewes treated with PTG.

Embryotoxicity of regressing corpora lutea in ewes.

Experiments were performed to test the null hypotheses that embryonic survival is not affected by the presence of regressing corpora lutea in progestogen-supplemented ewes, and that the embryotoxic effects of regressing corpora lutea do not act locally on embryos in the uterine horn adjacent to the regressing corpora lutea. In Expt 1, laparotomies were performed on day 4 after mating, and progestogen supplementation was initiated and continued until pregnancy was diagnosed by ultrasonography on day 25. On day 4 after mating, ewes were lutectomized (n = 17) or sham lutectomized (n = 15), and injected (i.m.) with 5 mg PGF2alpha at 8 h intervals for 2 or 3 days. Controls (n = 14) were sham lutectomized and injected with saline as described above. Pregnancy rates did not differ in ewes treated with PGF2alpha for 2 rather than 3 days. Pregnancy rates were lower in ewes treated with PGF 2a compared with controls (P < 0.01). In ewes treated with PGF2alpha, lutectomy resulted in an increase in pregnancy rates (59%) compared with ewes subjected to sham lutectomy (33%; P < 0.05). In Expt 2, progestogen supplementation was initiated in the morning of day 4 after mating and continued until pregnancy diagnosis on day 25. In the afternoon of day 4, one ovary selected at random was lutectomized in ewes (n = 34) with at least one corpus luteum on each ovary, and the uterine horns were isolated by ligation to impede intraluminal transfer of luteal or uterine products that might initiate embryonic death. On days 5-8 after mating, equal numbers of ewes were injected i.m. with either saline or 5 mg PGF2alpha at 8 h intervals. Pregnancy rates did not differ between isolated uterine horns contralateral and ipsilateral to the regressing corpus luteum; however, pregnancy rates were lower in PGF2alpha-treated ewes than in saline-treated ewes (34 and 77%; P < 0.05). In conclusion, regressing corpora lutea exert an embryotoxic effect; however, there is no evidence that this effect occurs through systemic pathways.

Prostaglandin F(2alpha) regulation of the bovine corpus luteum endothelin system during the early and midluteal phase.

Recent evidence in the cow suggests that endothelin-1 (ET-1) plays a role during prostaglandin (PG) F(2alpha)-induced luteal regression. We have examined the effects of treatment with PGF(2alpha) during the early and midluteal phases on three components of the endothelin system: endothelin-converting enzyme-1 (ECE-1), ET type A receptor (ET(A)), and ET-1 in the bovine corpus luteum (CL). Cyclic beef cows were injected (0 h) on Day 4 or 10 with either saline or the PGF(2alpha) analogue Lutalyse (15 mg). The CL were collected at 2 (n = 11), 10 (n = 23), 24 (n = 15), or 48 h (n = 12) after treatment. The cows in which CL were removed after 10 h comprised of two experimental groups. The first group (n = 11) received one injection; the second group (n = 12) received two injections, one at 0 h and one at 8 h. The cows in which CL were collected after 24 and 48 h received one injection every 8 h. Semiquantitative reverse transcriptase-polymerase chain reaction was used to evaluate the mRNA encoding ECE-1, ET(A), and ET-1. The ECE-1 and ET(A) proteins were evaluated by semiquantitative Western blot analysis. The ET-1 was the most likely component of the endothelin system target for PGF(2alpha) regulation during the midluteal phase. The ET(A) and ECE-1 genes were constitutively expressed in the Day 4 and Day 10 CL. A practical application of this observation is that it may be possible to target the ET-1 gene as a way to manipulate the luteolytic action of PGF(2alpha).