Improvement by ergonovine of sperm transport, fertilization and pregnancy rates in ewes in natural or prostaglandin-induced estrus

Eight experiments were conducted with 451 ewes to test effects of ergonovine, prostaglandin F2 alpha (PGF2 alpha) and phenylephrine on sperm transport and fertility. In most experiments, ewes were mated at estrus and necropsied 2 or 3 h later. Sperm were flushed from the oviducts, uterus and anterior, middle and posterior thirds of the cervix and counted. Various doses of PGF2 alpha or phenylephrine given im at mating caused no significant increase in sperm numbers in any segment of the tract 2 h later. Three different dose levels of ergonovine were given im to ewes in natural estrus 1 h after mating and ewes were necropsied 3 h after mating. Doses of .2 and 1.0 mg were ineffective, but .5 mg increased sperm numbers about 10-fold in the oviducts and uterus. When given im at the time of artificial insemination, .6 mg of ergonovine increased the fertilization rate at 3 d from 5/25 in control ewes to 12/25 (P less than .05). In three experiments with ewes in PGF2 alpha-induced estrus, .6 mg of ergonovine increased sperm numbers in the cervix and uterus at 3 h after mating and in the uterus and oviducts at 23 h, near ovulation. Other ewes were artificially inseminated in the external cervical os and one-half of the ewes were given .6 mg of ergonovine im; ewes not returning to estrus were laparotomized at 22 to 26 d and embryos removed. After insemination during natural estrus with .2 ml of semen, pregnancy rates were 14/25 for control ewes and 15/25 for ergonovine-treated ewes; after insemination during natural estrus with .1 ml of semen, 6/35 and 18/35 (P less than .005); after insemination during PGF2 alpha-induced estrus with .2 ml of semen, 7/60 and 12/60. Fertilization and pregnancy rates combined were 32/145 (22%) for all control ewes and 57/145 (39%) for ergonovine-treated ewes (P less than .005).     

Estrous cycle-dependent differences in responsiveness to prostaglandins and contractile agents in sheep (Ovis aries) cervical smooth muscle

We investigated the influence of the phase of the estrous cycle on mechanical responses elicited in sheep cervix by potassium chloride (KCl), acetylcholine chloride (ACh), prostaglandin F2 alpha (PGF2 alpha) and prostaglandin E1 (PGE1). The cervix of adult ewes (n = 48) were classified according to the presence or absence of corpora lutea (luteal or follicular phase, respectively). Muscle strips of the circular and longitudinal layers were prepared in an organ bath and coupled to an isometric force transducer. Concentration-response curves were obtained noncumulatively. KCl and ACh produced concentration-dependent contractions in all preparations in both phases of the estrous cycle. However, maximum effect, EC50 and slope values of KCl and ACh were not significantly different between muscle layers, as well as between the phases of the estrous cycle. The prostanoid, PGF2 alpha, produced a significant reduction in the amplitude of spontaneous contractions for all preparations. The depressant effect of PGF2 alpha on spontaneous contractions of circular smooth muscle was significantly greater during the follicular than the luteal phase, whilst the depressant effect of PGF2 alpha on the longitudinal layer did not differ between phases of the estrous cycle. PGE1 significantly reduced the amplitude of spontaneous contractions on circular but not on longitudinal preparations. In conclusion, we have characterized with in vitro preparations of circular and longitudinal muscle layers of ewes during the follicular and luteal phases of the estrous cycle, the parameters of the K- and ACh-induced contractions on cervix and the efficacy of PGF2 alpha and PGE1 on inhibition spontaneous contractile activity.     

Relationships between LH and estradiol-17 beta after removal of luteal progesterone in the ewe

Three experiments were conducted to examine the relationship between systemic concentrations of luteinizing hormone (LH) and estradiol-17 beta (E2) after withdrawal of progesterone in cycling ewes. In Exp. 1, ewes were assigned randomly to one of three treatments: laparotomy (C), removal of the luteal ovary (ULO), or ULO plus anesthesia with sodium pentobarbital for 6 h beginning 4 h after surgery. Anesthesia was used in an attempt to block the expected increase in tonic secretion of LH. Patterns of LH and E2 in these three groups did not differ during the 24-h experimental period. In Exp. 2, a longer period of anesthesia was utilized. Forty-eight ewes were assigned at random to one of four treatments: C, ULO, lutectomy or an intrafollicular injection of prostaglandin F2 alpha (PGF2 alpha). One-half of the ewes in each group were anesthetized with sodium pentobarbital from initiation of treatment (0 h) until 10 h after surgery. Sodium pentobarbital did not suppress the increases in LH and E2 after progesterone withdrawal. The regression of concentrations of E2 on concentration of LH was not significant. In Exp. 3, ewes were infused with either saline or dopamine after receiving an im injection of PGF2 alpha. Tonic secretion of LH increased after 4 h in ewes infused with saline, but not in ewes infused with dopamine. Despite the suppression of LH, concentrations of E2 increased in dopamine-treated ewes as in control ewes. Therefore, the initial increase in E2 after a decline of progesterone in cycling ewes is independent of increases in LH.     

Concentrations of ovarian and pituitary hormones following prostaglandin F2 alpha-induced luteal regression in ewes varies with day of the estrous cycle at treatment

Prostaglandin F2 alpha (PGF2 alpha) was injected on d 5, 8 or 11 postestrus in ewes to determine how stage of the estrous cycle would affect PGF2 alpha-induced changes in concentrations of ovarian and pituitary hormones and intervals to the onset of estrus and the preovulatory surge of luteinizing hormone (LH). Initial concentrations of progesterone and average values during the 12 h after PGF2 alpha were related positively to the day of cycle on which PGF2 alpha was administered. Patterns of decline in progesterone after injection of PGF2 alpha were similar among the 3 d. Concentrations of LH in plasma increased in a similar manner from 0 to 12 h in all ewes. After 12 h LH continued to increase, plateaued or declined in ewes treated on d 5, 8 or 11, respectively. Initial concentrations of follicle stimulating hormone (FSH) in plasma were related positively to day of treatment. After treatment with PGF2 alpha, FSH increased within 2 h on d 5 but declined by that time on d 8 or 11. Concentrations of estradiol following treatment did not vary with day. The onset of estrus and the preovulatory surge of LH occurred at 36 and 35, 40 and 45, and 48 and greater than 48 h in ewes treated on d 5, 8 or 11, respectively. It is concluded that: 1) the initial increase in LH is dependent on a decrease in plasma progesterone and 2) differences in patterns of secretion of gonadotropins before the preovulatory surge of LH might be caused by differences in progesterone or progesterone:-estradiol ratio when luteal regression is induced on different days of the estrous cycle.     

Effects of standing estrus and supplemental estradiol on changes in uterine pH during a fixed-time artificial insemination protocol

Cows that exhibit estrus within 24 h of fixed-time AI have elevated concentrations of estradiol and greater pregnancy rates compared with cows not in estrus. Our objective was to determine whether estradiol, estrus, or both had an effect on uterine pH during a fixed-time AI protocol. Beef cows were treated with the CO-Synch protocol (100 mircog of GnRH on d -9; 25 mg of PGF(2alpha) on d -2; and 100 mircog of GnRH on d 0). One-half of the cows received an injection of estradiol cypionate (ECP; 1 mg) 12 h after PGF(2alpha). Cows detected in standing estrus within 24 h of the second GnRH injection were considered to be in standing estrus. Uterine pH was determined in all animals 12, 24, and 48 h after the PGF(2alpha) injection. For Exp. 1, pH was also determined 72 and 96 h after the PGF(2alpha) injection; in Exp. 2, pH was also determined at 54, 60, 66, 72, 78, 84, 90, and 96 h after the PGF(2alpha) injection or until ovulation. A treatment x time interaction (P < 0.01) influenced concentrations of estradiol. All cows had similar (P > 0.15) concentrations of estradiol at the time of ECP administration, but after ECP treatment all cows treated with ECP and control cows that exhibited estrus had greater (P < 0.01) concentrations of estradiol compared with nontreated cows that did not exhibit estrus. In all animals, estradiol diminished 48 h after the PGF(2alpha) (time of the second GnRH injection), but ECP-treated cows, regardless of estrus, had elevated (P < 0.02) concentrations of estradiol compared with control cows. There was a treatment x time interaction (P < 0.001) on uterine pH. All cows had similar uterine pH (P > 0.19) 24 h after the PGF(2alpha) injection. Control cows that did not exhibit estrus had a greater uterine pH compared with control cows that exhibited estrus (P < 0.01) and ECP cows that exhibited estrus (P = 0.05) 48 h after the PGF(2alpha) injection (7.0 +/- 0.1 vs. 6.7 +/- 0.1 and 6.8 +/- 0.1, respectively). Estradiol cypionate-treated cows not exhibiting estrus were intermediate (6.8 +/- 0.1; P > 0.05). All cows had similar uterine pH 72 h after the PGF(2alpha) injection through ovulation (P > 0.06). In summary, uterine pH was similar among all animals that exhibited estrus, regardless of treatment with ECP.     

Maintenance of the corpus luteum of early pregnancy in the ewe. IV. Changes in luteal sensitivity to prostaglandin F2 alpha throughout early pregnancy

Two experiments were conducted to examine the temporal aspects of luteal resistance to the luteolytic effect of prostaglandin (PG) F2 alpha during early pregnancy. In Exp. 1, 14 pregnant and 12 nonpregnant ewes were treated with PGF2 alpha either on d 10 or 13 post-estrus. Jugular venous blood samples were collected at -30 min, 0, 6, 12, 18, 24, 30 and 36 h post-injection for quantification of progesterone. The difference (delta P) between pre-treatment and post-treatment concentrations of progesterone was calculated for each ewe. There was a significant interaction between pregnancy status and day of treatment on delta P (P less than .05). Pregnant and nonpregnant ewes treated on d 10 showed a large delta P. A large delta P also was observed in nonpregnant ewes treated on d 13 post-estrus. However, delta P in pregnant ewes treated on d 13 was smaller than in the other three groups (P less than .05). The temporal patterns of concentrations of progesterone in serum were different among treatment groups (P less than .05). A suppression in the concentration of progesterone was observed by 24 h post-injection in all four treatment groups. Progesterone returned to pre-treatment levels only in pregnant ewes treated on d 13. In Exp. 2, 47 pregnant ewes were treated with PGF2 alpha on d 10, 13, 16, 19, 22, 26 or 30 postestrus. Blood samples were collected and data were analyzed as described for Exp. 1.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of oxytocin, prostaglandin F2 alpha and reproductive tract manipulations on uterine contractility in Holstein cows on days 0 and 7 of the estrous cycle

The influence of various treatments on cattle at the time of natural estrus vs estrus induced by prostaglandin F2 alpha (PGF2 alpha) or at d 7 of the estrous cycle were studied; the latter is when embryo transfer often is performed. Eight lactating and 25 nonlactating, normal cycling cows were tested many times while in estrus and 7 d after estrus. A balloon was positioned in the body of the uterus to record changes in intrauterine pressure following clitoral massage, cervical massage, vaginal distention, electrical stimulation of the cervix and vagina, tailhead rubbing, udder massage and the injection of oxytocin or PGF2 alpha. Blood oxytocin and intramammary pressure were measured. There were no differences between cows in estrus spontaneously or those induced, so these groups were combined. Intravenous oxytocin injections of .5, 1, 2, 4 and 15 IU increased blood levels of oxytocin. Intramammary pressure was increased by all oxytocin doses, but greater than or equal to 2 IU were required to cause substantial changes in uterine contractions. As expected, the peak contractions during control periods for cows in estrus were high, averaging 31 mm Hg vs 11 mm Hg on d 7. None of the manipulations of the reproductive organs caused detectable oxytocin release or increases in intramammary pressure, contrasting to responses to massage of the udder. Clitoral massage increased peak uterine pressure by 32 to 60% in four experiments. It did not induce luteinizing hormone release. The contraction was immediate, was not sustained and could be obtained repeatedly, suggesting a reflex response. Treatment with PGF2 alpha increased intramammary pressure. It increased uterine pressure on d 7, but had no effect at estrus. This contrasts with injected oxytocin, which resulted in the highest amplitude for cows in estrus, although the response on d 7 was greater in proportion to the low activity in controls at that time. It is concluded that manual manipulation of the reproductive tract (other than clitoral massage) has little effect on uterine contractility.     

Effects of prostaglandin F2 alpha and pregnant mares' serum gonadotropin (PMSG) on the reproductive performance of fluorogestone acetate PMSG-treated ewes

Two experiments were conducted to examine the effect of prostaglandin F2 alpha (PGF2 alpha) and pregnant mares' serum gonadotropin (PMSG) on the reproductive performance (fertility, prolificacy and fecundity) of ewes previously treated with fluorogestone acetate (FGA) and PMSG. In the first experiment, 29 ewes were synchronized for estrus with FGA and PMSG but not bred at the postsynchronization estrus. On day 10 of the first post-synchronization estrous cycle, they were injected IM with 15 mg PGF2 alpha and 500 IU PMSG and exposed to experienced, fertile, raddled rams. Twenty-four of the 29 ewes (83%) had viable fetuses 9 weeks after the PGF2 alpha-PMSG treatment. In the second experiment, 64 ewes were treated with FGA-PMSG, and 33 were exposed to fertile, raddled rams at the synchronized estrus. A second group of 31 ewes was not bred at the synchronized estrus, but on day 12 of the postsynchronized estrous cycle, they were injected IM with 15 mg PGF2 alpha and 500 IU PMSG and exposed to fertile, raddled rams. Sixty-six percent of the 33 ewes lambed in the FGA-PMSG-treated group and 60% of the 31 ewes lambed in the PGF2 alpha-PMSG-treated group. Differences in reproductive performance between these two treatment groups were not statistically significant. The results suggest that the PGF2 alpha-PMSG treatment combination does not adversely affect reproductive performance of ewes.     

Effect of prostaglandin F2 alpha administration on early pregnancy in ewes

Two trials were conducted with ewes to determine the effects of prostaglandin F2 alpha (PGF) administration during the first week of gestation. In trial 1, ewes (n = 134) were checked for breeding activity once daily and half of them received 10 mg PGF im at either 0, 24, 48, 72, 96, 120 or 144 h after detection of a breeding mark. The other half served as uninjected controls. In trial 2, ewes (n = 153) were checked for breeding activity twice daily. Two-thirds of the ewes received 10 mg PGF at either 24, 36, 48, 60, 72, 84, 96, 108, 120 and 132 h following detection of a breeding mark. The other one-third of the ewes served as uninjected controls corresponding to treatment times of 24, 48, 72, 96 or 120 h. In trial 1, the percentage of ewes lambing as a result of first service decreased as time of administration of PGF increased. The first-service pregnancy rate was 87.5% for ewes given PGF at 0 h and 0% for ewes given PGF at 144 h. Fewer (P less than .05) ewes given PGF at 96, 120 or 144 h after first mating lambed than control ewes. Similarly in trial 2, fewer (P less than .05) ewes given PGF at 96, 108, 120 or 132 h after first mating lambed than did controls. The total number of ewes lambing as a result of the entire breeding season did not differ (P greater than .05) between treated and control ewes in trial 1 (88.2 vs 87.3%) or trial 2 (85.7 vs 83.3%).(ABSTRACT TRUNCATED AT 250 WORDS)     

Supplemental norgestomet,progesterone, or melengestrol acetate increases pregnancy rates in suckled beef cows after timed inseminations

In Exp. 1, 187 lactating beef cows were treated with injections of GnRH 7 d before and 48 h after prostaglandin F2alpha (PGF2alpha; Cosynch) or with Cosynch plus a 7-d treatment with an intravaginal progesterone (P4)-releasing insert (CIDR-B; Cosynch + CIDR). In Exp. 2, 183 lactating beef cows were treated with the Cosynch protocol or with Cosynch plus a 7-d treatment with norgestomet (Cosynch + NORG). In Exp. 1 and 2, blood samples for later P4 analyses were collected on d -17, -7 (first GnRH injection), 0 (PGF2alpha injection), and at timed artificial insemination (TAI; 48 h after PGF2alpha). In Exp. 3, 609 lactating beef cows were treated with the Cosynch + CIDR protocol or were fed 0.5 mg of melengestrol acetate (MGA) per day for 14 d before initiating the Cosynch protocol 12 d after the 14th d of MGA feeding (MGA + Cosynch). Blood samples were collected as in Exp. 1 and 2, plus additional samples on d -33 and -19 before PGF2alpha. In Exp. 4, 360 lactating beef cows were treated with a Cosynch + CIDR protocol, with TAI occurring at either 48 or 60 h after PGF2alpha, while receiving either GnRH or saline to form four treatments. Blood samples were collected as in Exp. 1 and 2. In Exp. 1, addition of P4 reduced the ability of the first GnRH injection to induce ovulation in anestrous cows with low P4 before PGF2alpha but improved (P = 0.06) pregnancy rates (61 vs 66%). In Exp. 2, the addition of NORG mimicked P4 by likewise increasing (P < 0.01) pregnancy rates (31 vs 51%) beyond those after Cosynch. In Exp. 3, the Cosynch + CIDR protocol increased (P < 0.001) pregnancy rates from 46 to 55% compared to the MGA + Cosynch protocol. In Exp. 4, administration of GnRH at TAI improved (P < 0.05) pregnancy outcomes (50 vs 42%), whereas timing of TAI had limited effects. We conclude that a progestin treatment concurrent with the Cosynch protocol improved pregnancy outcomes in all experiments, but pretreatment of cows with MGA was not as effective as the CIDR insert or NORG implants in this Cosynch-TAI model. Most of the improvement in pregnancy rates was associated with the increase in pregnancy rates of anestrous cows, regardless of whether ovulation was successfully induced in response to GnRH 7 d before PGF2alpha. Injection of GnRH at TAI following the Cosynch + CIDR protocol increased pregnancy rates in cycling cows with high P4 before the PGF2alpha injection and in anestrous cows with low P4 before PGF2alpha injection.     

Maintenance of the corpus luteum of early pregnancy in the ewe. III. Differences between pregnant and nonpregnant ewes in luteal responsiveness to prostaglandin F2 alpha

The effects of pregnancy and number of corpora lutea on luteal regression induced with prostaglandin F2 alpha (PGF2 alpha) were examined in 93 ewes. Bred and nonpregnant ewes were assigned randomly to receive a single im injection of PGF2 alpha: 0, 2, 4, 6, 8 or 10 mg/58 kg body weight. Injections were given on d 13 postestrus. The concentration of progesterone in serum 24 h after PGF2 alpha injection was affected by dose (P less than .001). The effect of pregnancy and the interaction of pregnancy with number of corpora lutea on levels of progesterone in serum were significant (P less than .05); therefore, data were partitioned according to pregnancy status and analyzed separately. There was an effect of number of corpora lutea on serum concentration of progesterone in pregnant (P less than .01) but not nonpregnant ewes (P greater than .10). Similar relationships among groups were observed for the concentration of progesterone in luteal tissue. In nonpregnant ewes the minimum dose of PGF2 alpha to produce a significant suppression of progesterone in serum (P less than .05) was 4 mg/58 kg body weight. In pregnant ewes with one or two corpora lutea, the minimum effective doses were 6 and 10 mg/58 kg body weight, respectively. The concentration of 13,14-dihydro-15-keto-prostaglandin F2 alpha (PGFM) in serum was related to the dose of PGF2 alpha injected. There were no differences in the concentration of PGFM in serum between pregnant and nonpregnant ewes either before or after injection. Corpora lutea of early pregnancy appear to be resistant to the luteolytic effect of PGF2 alpha.(ABSTRACT TRUNCATED AT 250 WORDS)     

FSH injections and intrauterine insemination in protocols for superovulation of ewes

In Exp. 1, five injections of crude FSH (20 mg total), starting 36 h before removal of progestin pessaries or injection of prostaglandin F2 alpha (PGF2 alpha), were found to be as effective for superovulation of ewes as seven injections starting 72 h before removal of pessaries. Seven injections caused superovulation (more than three ovulations) in 13 of 15 ewes with an average (all ewes) of 12 +/- 2.3 ovulations. Five injections caused superovulation in 17 of 19 ewes with an average of 11.5 +/- 1.6 ovulations after pessary-regulated estrus in 12 of 16 ewes with an average of 12.4 +/- 2.2 ovulations after PGF2 alpha injection. In Exp. 2, intrauterine artificial insemination (IUAI) of naturally mated ewes provided ovum fertilization rates of 75.3% with an embryo recovery rate of 39.4%, whereas mating alone (MATED) provided fertilization rates of 64.2% with an embryo recovery rate of 72.1%. However, subsequent replications of the MATED protocol (MATED2) resulted in a low fertilization rate (34.2%). In Exp. 3, surgical IUAI at 22 h after removal of pessaries resulted in a low rate of superovulation (5 of 12 ewes), presumably as the result of pentobarbital anesthesia. Conducting IUAI at 46 h after removal of pessaries resulted in a high rate of superovulation (14 of 16 ewes) but a low embryo recovery rate (30.7%). In two subsequent trials, IUAI at 40 h after removal of pessaries resulted in most ewes superovulating (33 of 35 in April/May and 16 of 17 in September) with acceptable embryo recovery (65.2% and 71.3%) and high fertilization rates (82.8% and 96.4%).(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of prostaglandin F2 alpha on the adenylate cyclase and phosphodiesterase activity of ovine corpora lutea

The objective of the present study was to determine the temporal relationships among luteal adenylate cyclase activity, luteal phosphodiesterase activity, luteal progesterone concentration and plasma progesterone concentration during prostaglandin F2 alpha (PGF2 alpha)-induced luteolysis in ewes. Corpora lutea (CL) were removed from cycling ewes on d 9 (d 0 = first day of estrus) at 0, 2, 4, 6, 12 and 24 h (seven to eight ewes/group) after PGF2 alpha administration (im). Jugular blood samples were collected at the time of enucleation of CL and analyzed for progesterone. Plasma and luteal progesterone concentrations were decreased (P less than .05) by 4 and 12 h after PGF2 alpha injection, respectively. Basal adenylate cyclase, luteinizing hormone (LH)-activated adenylate cyclase, guanylylimidodiphosphate [Gpp(NH)p]-activated adenylate cyclase and LH plus Gpp(NH)p-activated adenylate cyclase activities were decreased (P less than .05) by 2 h after PGF2 alpha injection. The decrease in adenylate cyclase activity paralleled the decrease in plasma progesterone concentration over time. Luteinizing hormone stimulated (P less than .05) adenylate cyclase activity relative to basal activity at 0, 2, 12 and 24 h post-PGF2 alpha; whereas, Gpp(NH)p stimulated (P less than .01) adenylate cyclase activity relative to basal activity at each time point. In contrast to the decrease in adenylate cyclase activity, phosphodiesterase activity was increased (P less than .05) at 2 and 4 h post-PGF2 alpha. These results suggest that a decrease in adenylate cyclase activity coupled with an increase in phosphodiesterase activity may decrease the intracellular adenosine 3',5' cyclic monophosphate (cAMP) concentration.(ABSTRACT TRUNCATED AT 250 WORDS)     

Secretory function of the ovine uterus: effects of gestation and steroid replacement therapy

In Exp. 1, changes in uterine fluid content of protein, calcium and prostaglandin F2 alpha (PGF) were examined between d 30 to 144 of gestation. Volume of uterine fluid (mean +/- SE) in the nongravid uterine horn of unilaterally pregnant ewes increased (P less than .05) between d 30 (8 +/- 1 ml) and d 144 (749 +/- 46 ml) of gestation. Protein concentration and total protein in uterine fluid also increased (P less than .05) between d 30 (2.3 +/- .4 mg/ml; 19 +/- 7 mg) and d 144 (23.5 +/- 3.3 mg/ml; 17.4 +/- 2.0 g). Total recoverable calcium (mg) in secretions increased from .1 mg (d 30) to over 1.4 g (d 144) due to day of gestation effects (P less than .05). Total PGF also increased (P less than .01) from 7 ng on d 30 to 15.7 g on d 144. In Exp. II, ovariectomized (OVX) ewes were treated with either corn oil, estrone (E), progesterone (P) or P+E (PE) for 30 d. Treatment with P or PE increased calcium concentration and content (P less than .01) in secretions, but differences in uterine fluid volumes and concentrations of protein and PGF were not significant. Treatment of OVX ewes with P stimulated (P less than .05) in vitro synthesis of secretory proteins by endometrium, whereas treatment with E enhanced release of unlabeled proteins (P less than .05). The major endometrial secretory proteins were identified in allantoic fluids from d 60 to term and were detected along the mesenchymal border of the chorioallantois using immunohistochemistry. Results from this study indicate that P may be a primary hormone regulating accumulation of fluid, protein, calcium and specific endometrial proteins in the uterine lumen during gestation, and that uterine milk proteins gain access to the fetal-placental unit.     

Active immunization of heifers against prostaglandin F2 alpha: potential as a sterilization vaccine

Four experiments were conducted with 210 heifers in an attempt to develop a sterilization vaccine by active immunization against prostaglandin F2 alpha (PGF2 alpha) and to evaluate feedlot performance following immunization against the combination of PGF2 alpha and estrogens. The objectives were: development of a PGF2 alpha-ovalbumin conjugate that would induce antibody formation when used with complete Freund's adjuvant (CFA); comparison of CFA with other adjuvants in relation to PGF2 alpha antibody binding and maintenance of the corpus luteum; examination of growth performance in immunized heifers against both PGF2 alpha and estradiol-17 beta and evaluation of sterilization in PGF2 alpha-immunized heifers maintained with bulls. A PGF2 alpha-ovalbumin conjugate was developed that resulted in antibody production against PGF2 alpha, although antibody binding was quite low. The antibody response in heifers was higher in Exp. I and II than in Exp. III and IV (P less than .05). Complete Freund's adjuvant was the best adjuvant in inducing antibody formation compared with all other adjuvants tested (P less than .01). Corpus luteum (CL) function was maintained for 2.5 mo and ovulation was apparently blocked in Exp. I. The results of Exp. II confirmed those of Exp. I. Fewer than half of the heifers in Exp. III and IV had prolonged estrous cycles. In Exp. IV, immunized heifers became sterile for at least 4 mo when kept with bulls, although the success rate was only 37%. The low levels of antibody titers to PGF2 alpha in Exp. III and IV may be the reason for the failure to maintain CL function in some heifers.(ABSTRACT TRUNCATED AT 250 WORDS)     

Changes in bovine luteal progesterone metabolism in response to exogenous prostaglandin F(2alpha)

An experiment was conducted to determine the effect of prostaglandin F2alpha (PGF2alpha) on luteal synthesis of progesterone (P4) and related progestins. Sixteen beef heifers were assigned in equal numbers to four groups in a 2x2 factorial arrangement of treatments. The experiment consisted of two levels of PGF2alpha analog (0 and 500 microg) and two levels of time (4 and 24 h after injection) of corpus luteum collection. All heifers were injected intravenously with saline (2 ml) or PGF2alpha (cloprostenol) on day 8 of the estrous cycle (estrus=day 0). Jugular blood was collected at 0, 1, 2, 3, 4 and 20, 21, 22, 23, and 24 h after injection. Resulting sera were analyzed for P4 by use of radioimmunoassay. Luteal tissue was analyzed by gas chromatography/mass spectrometry for P4, 20beta-hydroxyprogesterone, pregnenolone, and allopregnanolone (3beta-hydroxy-5alpha-pregnan-20-one). Treatment with PGF2alpha reduced serum concentrations of P4 as early as 1 h after injection (P<0.005) and steroid levels remained low over 24 h. Similarly, administration of PGF2alpha caused a decline in luteal P4 (P<0.005), 20beta-hydroxyprogesterone (P<0.10), and pregnenolone (P<0.05). In contrast, treatment with PGF(2alpha) caused an increase in luteal allopregnanolone over time (time x treatment interaction; P<0.05). These data are interpreted to suggest that PGF2alpha promotes conversion of P4 to the metabolite allopregnanolone.     

Influence of hormone supplementation to extended semen on artificial insemination,uterine contractions,establishment of a sperm reservoir,and fertility in swine

This study was performed to quantify the effect of hormone addition to semen using a low-fertility model to evaluate its effectiveness and mode of action. At 24 h after the onset of estrus, all gilts received a single low-dose AI (0.5 x 10(9) sperm/80 mL) with no hormone (control, C), estrogens (E, 11.5 microg), PGF2alpha (PG, 5 mg of Lutalyse), or oxytocin (OT, 4 IU), which were then evaluated for semen backflow (n = 48), oviductal and uterine sperm numbers (n = 28), uterine contractions (n = 12), pregnancy rate (PR, n = 120), and number of fetuses (n = 67). In Exp. 1, backflow of semen from the uterus was collected for 8 h after AI, whereas PR and fetuses were assessed at d 25 to 30 after AI. In Exp. 2, backflow was collected and reproductive tracts flushed to determine sperm numbers in the oviducts and the anterior segments of the uterus. In Exp. 3, sows were monitored for uterine contractions for 1 h before AI and for 2 h after AI. In Exp. 1, there was a treatment x time interaction for fluid loss (P < 0.001), but by 8 h after AI, there was no difference in the total volume (70 +/- 1 mL) of semen lost between hormone treatments (85%) compared to controls (90%). There was also a treatment x time interaction (P < 0.05) for number of sperm lost in the backflow (2.1 +/- 0.1 x 10(8)), but by 8 h following AI, there was no effect on total sperm lost for the hormone treatments (38%) compared to C (54%). There was a trend (P = 0.10) for increased numbers of sperm in the uteri of hormone-treated gilts (6.0 +/- 1.3 x 10(4)) compared with C gilts (2.2 +/- 1.3 x 10(4), but there was no effect of treatment on sperm numbers in the oviducts (3.2 +/- 1.3 x 10(4)). Within 0.5 h of AI, there was an increase in the frequency of contractions for PG compared with the other treatments (14.2 vs. 6.3/h, P < 0.005), however there was no effect on amplitude (54 mmHg) or duration (35 s) of contractions. The PR was not influenced by treatment and averaged 54% (P > 0.60), but total numbers of healthy fetuses were increased (P < 0.04) by PG (8.7) and tended (P = 0.06) to be increased for OT (8.4), but not for E (7.2) compared to C (5.8). Hormone addition to semen increased numbers of fetuses and this may be related to an alteration in the pattern of fluid and sperm loss after AI and a tendency for increased numbers of sperm in the anterior segment of the uterus. Therefore, in situations of lowered fertility, hormone addition could be a strategy to limit infertility in swine.     

Putative urinary pheromone of bulls involved with breeding performance of primiparous beef cows in a progestin-based estrous synchronization protocol

The objective of this study was to determine if factors associated with the biostimulatory effect of bulls alter breeding performance of primiparous, suckled beef cows using a progestin-based estrous synchronization protocol. We tested the hypotheses that the estrous synchronization response and AI pregnancy rates differ among cows exposed to bulls, continuously exposed to bull urine, and exposed to fence-line contact with bulls or cows not exposed to bulls or bull urine. Data were collected from 3 experiments performed over consecutive years. Cows were assigned to the following treatments: bull exposure (BE; n = 26) or no bull exposure (NB; n = 25) in Exp. 1, bull urine exposure (BUE; n = 19) or steer urine exposure (SUE; n = 19) in Exp. 2, and fence-line contact with bulls (BFL; n = 26) or no bull exposure (NB; n = 26) in Exp. 3. Synchronization protocols in each experiment included the use of a controlled internal drug release device (d -10), PGF(2alpha) (d -3), and GnRH and fixed-time AI (TAI; d 0). Cows that were observed in estrus by 60 h after PGF(2alpha) were inseminated 12 h later. Cows not observed in estrus by 60 h after PGF(2alpha) were TAI at 72 h and given GnRH (100 mug). Pregnancy was determined by ultrasonography 35 d after TAI. In Exp. 1, 2, and 3, cows were exposed directly to bulls, bull urine, or bull fence-line contact for 35, 64, and 42 d, respectively. Data were analyzed between treatments within each experiment. The proportion of estrous cycling cows did not differ between treatments at the beginning of each experiment; however, more (P < 0.05) BE and BFL cows were estrous cycling at the beginning of the estrous synchronization protocol than NB cows in Exp. 1 and 3. The proportion of cows that showed estrus and interval to estrus after PGF(2alpha) did not differ between treatments in Exp. 1 and 3. However, in Exp. 2, more BUE cows tended (P = 0.09) to have shorter intervals to estrus and to exhibit estrus after PGF(2alpha) than SUE cows. Overall, AI pregnancy rates were greater (P < 0.05) for BE and BUE cows than for NB and SUE cows in Exp. 1 and 2, respectively. There was no difference in AI pregnancy rates between BFL and NB cows in Exp. 3. The presence of bulls and exposure to bull urine appeared to improve breeding performance of primiparous beef cows using a progestin-based estrous synchronization protocol, whereas fence-line bull exposure was insufficient to cause this biostimulatory effect. We propose that a novel urinary pheromone of bulls may be responsible for the enhancement of fertility in the primiparous, postpartum cow.     

Effects of feed restriction on reproductive and metabolic hormones in ewes

The goal of this study was to determine the effects of short-term feed withdrawal on reproductive and metabolic hormones during the luteal phase of the estrous cycle in mature ewes. Mature ewes observed in estrus were assigned randomly to control and fasted groups (n = 10 per group Trials 1 and 2). For Trials 1 and 2, control ewes had ad libitum access to feed, whereas fasted ewes were not fed from d 7 through 11 of their estrous cycle; on d 12, all ewes were treated with 10 mg of PGF2alpha, and fasted ewes were gvien ad libitum access to feed. For Trial 1, blood samples were collected daily through fasting and at 2-h intervals following PGF2alpha for 72 h. Serum concentrations of insulin (P < or = 0.002) and IGF-I (P < or = 0.01), but not GH (P > or = 0.60), were decreased during fasting compared with fed ewes. Serum concentrations of 29 (P = 0.02) and 34 kDa (P = 0.04) IGFBP were greater in fasted ewes at 96 h after initiation of fasting than in control ewes. Two control and four fasted ewes in Trial 1 did not exhibit a preovulatory surge release of LH by 72 h. Therefore, Trial 2 was conducted so that the timing of the LH surge could be predicted following the collection of blood samples at 2-h intervals for 112 h and then at 6-h intervals until 178 h following PGF2alpha administration and realimentation. The magnitude of the preovulatory LH surge in Trial 2 was decreased (P = 0.009) and delayed (P = 0.04), and serum concentrations of estradiol were diminished (P < or = 0.03) 12 h before the LH surge in fasted ewes. Ovulation rates were not influenced (P > or = 0.32) by fasting in Trials 1 and 2. Serum concentrations of progesterone in both Trials 1 and 2 were, however, greater (P < 0.001) in fasted than in control ewes. A third trial with ovariectomized ewes was conducted to determine whether the increased serum concentrations of progesterone observed in fasted ewes during Trials 1 and 2 were ovarian-derived. Ovariectomized ewes were implanted with progesterone-containing intravaginal implants and allotted to control (n = 5) or fasted (n = 5) treatment groups and fed as described for Trials 1 and 2. Similar to intact ewes, serum concentrations of progesterone were approximately twofold greater (P < 0.001) in fasted than in control implanted ovariectomized ewes. In summary, feed withdrawal for 5 d during the luteal phase of the estrous cycle increased serum concentrations of progesterone and evoked endocrine changes that could perturb the subsequent estrous cycle.