Administration of recombinant bovine interferon-alpha I at the time of maternal recognition of pregnancy inhibits prostaglandin F2 alpha secretion and causes luteal maintenance in cyclic ewes.

The antiluteolytic protein, ovine trophoblast protein-1, which is secreted by sheep embryos at about the time of the maternal recognition of pregnancy, exhibits significant structural homology with alpha interferons. Experiments were conducted to examine the effects of intra-uterine and systemic administration of a recombinant bovine interferon-alpha I (rboIFN-alpha I) upon the interoestrus interval, endometrial oxytocin receptor concentrations and secretion of prostaglandin (PG) F2 alpha in cyclic ewes. In Expt 1, each ewe had a cannula placed in the tip of a uterine horn ipsilateral to a corpus luteum, 7 days after an induced oestrus. From day 9 after oestrus until day 19, ewes received either 200 (n = 4), 667 (n = 5) or 2000 (n = 9) micrograms/24 h of rboIFN-alpha I, meclofenamic acid (n = 4) or vehicle (n = 11). Other ewes received 2000 micrograms rboIFN-alpha I/24 h (n = 5) between days 12 and 15 only. All ewes were killed on day 19. Mean luteal phase, as determined by daily plasma progesterone measurements, was significantly longer (P less than 0.01) and mean concentrations of 13,14-dihydro-15-keto PGF 2 alpha (PGFM) in plasma were lower (P less than 0.05) in ewes receiving 667 or 2000 micrograms rboIFN-alpha I between days 9 and 19, or 2000 micrograms between days 12 and 15, than in animals from other treatment or control groups. A similar protocol was used in Expt 2, in which further ewes received either 2000 micrograms rboIFN-alpha I/24 h (n = 5) or vehicle (n = 5) by bolus infusions twice a day into one uterine horn. Mean luteal phase was significantly (P less than 0.05) longer in treated than in control animals, but differences in PGFM concentrations were not significant. In Expt 3, after a synchronized oestrus, ewes received either 2.5 mg rboIFN-alpha I by i.m. injection twice a day between days 12 and 15 (n = 10), 2.5 mg rboIFN-alpha I by i.m. injection twice a day between days 9 and 15 (n = 11), i.m. injection of vehicle alone twice a day (n = 20), or continual intra-uterine infusion of 2 mg rboIFN-alpha I/day between days 12 and 15 (n = 7). The mean luteal phase of ewes receiving rboIFN-alpha I by intrauterine infusion or i.m. injection between days 9 and 15 was significantly longer than for animals from the other two groups (P less than 0.05).(ABSTRACT TRUNCATED AT 400 WORDS)     

Effect of injected bovine interferon-alpha I1 on estrous cycle length and pregnancy success in sheep

In Exp. 1 twice daily i.m. injections of 2 mg recombinant bovine IFN-alpha I1 (rboIFN-alpha I1) (N = 24) or placebo (N = 25) were administered to ewes from Day 12 to Day 16 during a normal oestrous cycle. Treatment did not increase (P greater than 0.10) oestrous cycle length (20.7 +/- 1.2 versus 18.5 +/- 1.4 days). In Exp. 2, ewes were injected twice daily with 2 mg IFN (N = 34) or placebo (N = 36) from Days 11 to 18 after natural mating. The rboIFN-alpha I1 significantly (P = 0.05) improved pregnancy rate (79% versus 58%) as determined by a failure of ewes to return to oestrus within 50 days. The number of ewes that lambed was greatest in the rboIFN-alpha I1-treatment group (71% versus 50%; P = 0.07), and no teratogenic effects were observed in the young born to IFN-treated ewes. The study was repeated a second year with a more fecund group of ewes (Exp. 3). More (P = 0.08) ewes injected with rboIFN-alpha I1 (58/65) than placebo-treated ewes (48/61) were judged pregnant by ultrasound. Again more ewes lambed (55 versus 45) and more lambs were born (98 versus 80) from the rboIFN-alpha I1-treated group. Combining the data from both studies revealed a significant (P = 0.01) effect of treatment. The amount of antiviral activity in jugular vein blood of ewes injected with rboIFN-alpha I1 (2 mg) was determined over time in Exp. 4. Activity rose to a maximum (approximately 450 IRU/ml) within 1-2 h and declined by over 75% in 24 h. Single injections of 1, 2 and 5 mg in buffer or 2 mg emulsified in sesame oil all gave similar profiles of antiviral activity in jugular blood over a 48-h period. In Exp. 5, antiviral activity was measured in uterine vein, ovarian artery and jugular vein serum of untreated pregnant (N = 7) and non-pregnant (N = 11) ewes at Day 15 after mating. Activity was detected in the uterine vein (58 +/- 19 IRU/ml) of all pregnant ewes. The observations in Exps 1-5 are consistent with a role for conceptus-derived IFN-alpha in maternal recognition of pregnancy and suggest that supplemental IFN-alpha might be useful in improving pregnancy success in sheep.     

hCG treatment on day of mating improves embryo viability and fertility in ewe lambs

An investigation was conducted to determine the effects of supplementing hCG at mating time on luteal function, conceptus growth, placentation and reproductive performance in TexelxClun Forest ewe lambs. After oestrus synchronisation with progestagen sponges and PMSG ewe lambs were treated either with normal saline (n=102) or 150 IU of hCG (n=105) at mating time. At 25 days after mating 24 animals were slaughtered from each group to determine embryo viability and placental development. hCG significantly (P<0.05) increased crown-rump length (saline: 11.9+/-0.2 mm; hCG: 12.7+/-0.2 mm), amniotic sac width (saline: 11.4+/-0.4 mm; hCG: 12.0+/-0.3 mm) and the number of placentomes (saline: 90.8+/-7.3; hCG=122.4+/-6.3). Among the pregnant animals that were slaughtered on 25 days post-mating, ovulation rate tended to be higher in the hCG group compared to controls (saline: 1.16; hCG: 1.54). However, no difference was observed either in mean plasma progesterone concentrations (saline: 4.6 ng/ml; hCG: 4.9 ng/ml; sed 0.56) or in progesterone production from luteal slices when cultured in vitro (saline: 239.6+/-11.8 ng/mg; hCG: 263.2+/-13.6 ng/mg) between controls and hCG treated animals. Reproductive performance was also recorded in ewe lambs that were either treated with saline (n=78) or hCG (n=81). The total number of lambs born (saline: 38; hCG: 58) was significantly (P<0.05) greater in hCG group compared to saline-treated controls. Both lambing percentage (saline: 36%; hCG: 48%) and litter size (saline: 1.35; hCG: 1.48) tended to be greater (P<0.10) in hCG-treated animals compared to the controls. In conclusion, these data suggest that treatment of ewe lambs with hCG at the time of mating improves conceptus growth, placentation and number of lambs born.     

The synchrony of prostaglandin-induced estrus in cows was reduced by pretreatment with hCG

The induction of optimal synchrony of estrus in cows requires synchronization of luteolysis and of the waves of follicular growth (follicular waves). The aim of this study was to determine whether hormonal treatments aimed at synchronizing follicular waves improved the synchrony of prostaglandin (PG)-induced estrus. In Experiment 1, cows were treated on Day 5 of the estrous cycle with saline in Group 1 (n = 25; 16 ml, i.v., 12 h apart), with hCG in Group 2 (n = 27; 3000 IU, i.v.), or with hCG and bovine follicular fluid (bFF) in Group 3 (n = 21; 16 ml, i.v., 12 h apart). On Day 12, all cows were treated with prostaglandin (PG; 500 micrograms cloprostenol, i.m.). In Experiment 2, cows were treated on Day 5 of the estrous cycle with saline (3 ml, i.m.) in Group 1 (n = 22) or with hCG (3000 IU, i.v.) in Group 2 (n = 20) and Group 3 (n = 22). On Day 12, the cows were treated with PG (500 micrograms in Groups 1 and 2; 1000 micrograms in Group 3). Blood samples for progesterone (P4) determination were collected on Day 12 (Experiment 1) or on Days 12 and 14 (Experiment 2). Cows were fitted with heat mount detectors and observed twice a day for signs of estrus. Four cows in Experiment 1 (1 cow each from Groups 1 and 2; 2 cows from Group 3) had plasma P4 concentrations below 1 ng/ml on Day 12 and were excluded from the analyses. In Experiment 1, cows treated with hCG or hCG + bFF had a more variable (P = 0.0007, P = 0.0005) day of occurrence of and a longer interval to estrus (5.9 +/- 0.7 d, P = 0.003 and 6.2 +/- 0.8 d, P = 0.005) than saline-treated cows (3.4 +/- 0.4 d). The plasma P4 concentrations on Day 12 were higher (P < 0.0001) in hCG- and in hCG + bFF-treated cows than in saline-treated cows (9.4 +/- 0.75 and 8.5 +/- 0.75 vs 4.1 +/- 0.27 ng/ml), but there was no correlation (P > 0.05) between plasma P4 concentrations and the interval to estrus. In Experiment 2, cows treated with hCG/500PG and hCG/1000PG had a more variable (P = 0.0007, P = 0.002) day of occurrence of and a longer interval to estrus (4.2 +/- 0.4 d, P = 0.04; 4.1 +/- 0.4 d, P = 0.03) than saline/500PG-treated cows (3.2 +/- 0.1 d). The concentrations of plasma P4 on Days 12 and 14 of both hCG/500PG- and hCG/1000PG-treated cows were higher (P < 0.05) than in saline/500PG-treated cows (7.3 +/- 0.64, 0.7 +/- 0.08 and 7.7 +/- 0.49, 0.7 +/- 0.06 vs 5.3 +/- 0.37, 0.5 +/- 0.03 ng/ml). The concentrations of plasma P4 on Days 12 or 14 and the interval to estrus were not correlated (P > 0.05) in any treatment group. The concentrations of plasma P4 on Days 12 and 14 of hCG/500PG- or hCG/1000PG-treated cows were correlated (r = 0.65, P < 0.05; r = 0.50, P < 0.05). This study indicated that treatment of cows with hCG on Day 5 of the estrous cycle reduced the synchrony of PG-induced estrus and that this reduction was not due to the failure of luteal regression.     

Mid-gestation pregnancy termination by the progesterone antagonist aglepristone in queens

The efficacy of aglepristone, a progesterone receptor antagonist, to induce abortion on days 25 and 26 after first mating was investigated in queens. The cats were divided into two groups: aglepristone (10 mg/kg, subcutaneously) was injected twice, 24 h apart, on days 25 and 26 after first mating, into group I queens (n = 23). Group II queens (n = 6) were not treated and served as controls. Termination of pregnancy and expulsion of the fetuses were successful in 20 (87%) queens in group I. The mean interval between the first administration of aglepristone and the beginning of vaginal discharge was 5+/-1 days (range 4-7 days) and the mean duration of abortion, defined as time span from first occurrence of vaginal discharge to expulsion of all fetuses observed by ultrasonography was 1 day in nine cats, 2 days in five cats and in five cats, less than 1 day. Treatment failed in three queens. In one queen treatment resulted in birth (66 days after mating) of two vital kittens. In another case, three macerated fetuses were found intrauterine without vaginal discharge. In one cat, two fetuses were expulsed and two remained intrauterine and were born 66 days after last mating. All group II queens gave birth to vital kittens after a normal pregnancy length. The mean serum P4 concentrations were similar in treated and control animals. The results indicate that aglepristone treatment at day 25 of pregnancy could induce abortion in 87% of the treated queens. Itching at the site of injection right after injection was the only side effect noticed and only in one queen.     

Modification of follicular dynamics by exogenous FSH and progesterone, and the induction of ovulation using hCG in postpartum beef cows

Follicular growth and ovulation in response to FSH, progesterone and hCG were evaluated in postpartum beef cows. In Experiment 1, on Day 21 post partum, cows received an injection of either saline (control; n = 6), FSH (200 mg; n = 6), or a PRID (n = 5) for 10 d. Both FSH and PRID prolonged maintenance of a dominant follicle (15.5 +/- 1.16 and 14.4 +/- 1.29 d, respectively, vs 8.4 +/- 1.22 d in control; P < 0.01), and increased the maximum diameter of the dominant follicle (14.0 +/- 0.91 and 16.4 +/- 1.01 mm, respectively, vs 10.9 +/- 0.95 mm in control; P < 0.05). The PRID-maintained dominant follicle ovulated in 60% of cows, followed by normal estrous cycles (vs 0% in control; P = 0.01), whereas the dominant follicle ovulated in 33% of FSH-treated cows (P = 0.08). The PRID regimen shortened the interval to first ovulation preceding a normal cycle and continued cyclicity (44 +/- 4.1 vs 60 +/- 4.4 d in control; P = 0.02). In Experiment 2, on Day 21 post partum, cows received either saline (control), saline + PRID, or FSH + PRID (n = 16/group). Sixty hours after PRID withdrawal, cows received either saline or hCG (1,500 IU, n = 8/treatment). The FSH + PRID regimen increased the number of large (> 10 mm in diameter) follicles (3.6 +/- 0.43 vs 1.9 +/- 0.39 in control; P = 0.005). Both PRID and FSH + PRID prolonged maintenance of the largest follicle (11.0 +/- 0.82 and 11.2 +/- 0.91 d, respectively, vs 8.7 +/- 0.81 d in control; P < 0.05). The PRID-maintained dominant follicle ovulated in 50% of cows, followed by normal estrous cycles. The FSH + PRID-maintained largest follicle had become atretic at PRID withdrawal and was anovulatory. The FSH + PRID + hCG regimen increased the incidence of ovulation preceding a cycle of normal duration and continued cyclicity (100 vs 50% in PRID; P = 0.03), and reduced the interval to first ovulation preceding a cycle of normal duration and continued cyclicity (38 +/- 6.5 vs 58 +/- 6.3 d in control; P = 0.04). The area under the progesterone curve during the induced cycle was reduced after (PRID +/- FSH) + hCG than after PRID +/- FSH (P = 0.002). These results indicate that PRID alone or with FSH/hCG has the potential to modify the dominant follicle and initiate cyclicity in postpartum beef cows.     

Induction of the synthesis of the pregnancy-specific protein p70 in the endometrium by intramuscular injection of recombinant bovine interferon-alpha I1 in nonpregnant ewes.

We examined the effect of recombinant bovine interferon-alpha I1 (rboIFN-alpha I1) or recombinant bovine trophoblast protein-1 (rbTP-1) on protein synthesis by endometrial explants from Day-13 cyclic ewes and studied the ability of rboIFN-alpha I1 injected i.m. to influence subsequent protein secretion by endometrial tissue explants. In Expt 1, ewes were injected with either 2 mg rboIFN-alpha I1 or vehicle alone at 12 h intervals beginning on Day 11 of the oestrous cycle and ending on the morning of Day 13; 8 h after the last injection, ewes were hysterectomized and endometrial explant cultures were prepared. Explants were cultured for 24 h in leucine-deficient medium supplemented with 250 microCi L-[3H]leucine per culture. For Expt 2, additional explants were prepared from Expt 1 controls. Explants were cultured in the presence of 0, 20 or 200 ng/ml of either rboIFN-alpha I1 or rbTP-1 for 24 h in leucine-deficient medium supplemented with 250 microCi L-[3H]leucine per culture. Secreted proteins were analysed by two-dimensional electrophoresis and fluorography. There was a marked enhancement of a 70 kDa acidic protein, p70, in explants cultured in the presence of rboIFN-alpha I1 or rbTP-1. This polypeptide is a product of the gravid uterine horn from Day 14 to Day 20 of pregnancy and is a useful marker of the action of interferon-alpha (IFN-alpha) on endometrium. Enhanced production of p70 also occurred in ewes injected i.m. with rboIFN-alpha I1.(ABSTRACT TRUNCATED AT 250 WORDS)     

Intrauterine infusion of highly enriched bovine trophoblast protein-1 complex exerts an antiluteolytic effect to extend corpus luteum lifespan in cyclic cattle

Intrauterine infusion of enriched bovine trophoblast protein-1 complex (bTP-1) resulted in extension of interoestrous intervals and corpus luteum function in cyclic cattle. Conceptus proteins were obtained by culture of Day 17-18 conceptuses for 72 h. Media from the first (n = 28), second (n = 26) and third (n = 19) 24 h of conceptus incubations were utilized. A highly enriched preparation of bTP-1 was obtained by a combination of ammonium sulphate precipitation, ion-exchange chromatography, and h.p.l.c. gel filtration. Degree of purity of the final preparation was confirmed by gel electrophoresis and immunoblotting with antiserum to ovine trophoblast protein-1. Jersey cattle (3 per group) received intrauterine infusions, twice daily from Day 15.5 to 21.0, of bovine serum albumin, the entire array of bovine conceptus secretory proteins (bCSP) from the 3 days of conceptus culture, or bTP-1. Infusions were via a catheter into the uterine horn ipsilateral to the corpus luteum. Oestrous cycle length in bTP-1-treated cows (26.1 +/- 1.3 days) was greater than for cows given BSA (19.5 +/- 1.3 days) or bCSP (21.5 +/- 1.3 days). Similarly, progesterone concentrations in serum remained elevated for a longer period of time for bTP-1-treated cows than for cows treated with BSA or bCSP. Residual variance associated with vena cava concentrations of PGF-2 alpha at Days 19-21 after oestrus (which included the variance between 15-min periods within cows) was reduced in cows treated with bTP-1 as compared to other groups. Lack of a bCSP effect may have been due to low amounts of bTP-1 in conceptus-conditioned medium from cultures of greater than 24 h. None the less, purified bTP-1 was effective in extending luteal function and appears to be the antiluteolytic agent of early pregnancy.     

Circulating gonadotrophins and follicular dynamics in anestrous ewes after treatment with estradiol-17beta.

Plasma FSH, LH, estradiol (E2) and progesterone (P4) profiles and patterns of follicular growth and regression by ultrasonography were determined after E2 treatment (1 microg/kg) in anestrous ewes. Fifteen ewes were treated with one (group I, n=7) or two (group II, n=4) i.m. injections of E2 with a 24h interval, or two oil injections with a 24h interval (group C, n=4). Blood samples for E2, P4, FSH and LH determinations were collected daily 4 days before the initiation of the treatment (day 0), when bleeding increased to every 2h starting 2h before treatment until 56h after the first injection and from then on every 6h until day 8, and twice per day till the end of the experiment (day 9). During the experimental period (days -4 to 9), transrectal ultrasonic examinations were carried out daily using a 7.5 MHz linear array probe. Number and size of follicles > or =3mm in diameter were recorded. No estrous was detected before, during or after treatment. LH and FSH surges were observed 10-18h after the first E2 injection. The second E2 injection stimulated another release of LH but no surges. E2 inhibited FSH levels before the surge and the second E2 injection induced a longer inhibition. No ovulation was detected by ultrasonography during the experimental period and P4 levels remained low (<0.7 nmol/l) before, during and after the treatment in all ewes. There was an effect of E2 treatment on the diameter of the largest follicle, a decrease could be observed 3 days after the first injection in both ewes of groups I and II. The E2-treated groups had a higher frequency of ewes showing wave emergence on day 3 (day 1.5+/-1,2.4+/-0.4 and 2.5+/-0.5 for control, groups I and II). LH and FSH surges were observed after E2 treatment, but were not able to provoke ovulation neither luteinization. In contrast, the treatment was associated with the regression of the largest follicle and with emergence of a new follicular wave on day 3.     

Effect of eCG on early resumption of ovarian activity in postpartum dairy cows

The purpose of the present study was to hasten the resumption of ovarian activity early postpartum in lactating dairy cows, using equine chorionic gonadotropin (eCG), to enhance follicular growth, followed by hCG, to induce ovulation. Primiparous Holstein dairy cows (n=21) were assigned equally into eCG, eCG-hCG and Control groups. Cows in the eCG and eCG-hCG groups received an i.m. injection of eCG (500 IU Folligon?) on Day 6 postpartum. Cows in the eCG-hCG group were also given an i.m. injection of hCG (500 IU Chorulon?), once dominant follicle reached the diameter of 13-16 mm following eCG injection. Cows in Control group did not receive any treatment. Daily blood sampling and ultrasound examination were conducted, starting at Day 6 postpartum until confirming the third ovulation. Follicles ≥10 mm in diameter were detected on Day 11.5±1.48, 10.1±0.52 and 11.1±1.36 after calving in Control, eCG and eCG-hCG groups, respectively (P>0.05). The first wave dominant follicle ovulated in 71.4% of cows treated with eCG and eCG-hCG. In contrast, none of the first wave dominant follicles ovulated in Control cows. By Day 20 postpartum, all cows in eCG group, 6/7 cows in eCG-hCG group and none of the cows in Control group ovulated (P<0.05). Short estrous cycles (≤16 days) were detected in 2/7, 1/7 and 6/7 cows in eCG, eCG-hCG and control groups, respectively (P<0.05). In conclusion, injection of eCG on Day 6 postpartum could assist the early resumption of ovarian activity by enhancing ovarian follicle growth and early ovulation in postpartum cows. In this context, subsequent hCG injection may not provide any more beneficial effect.     

Steroid priming shortens prostaglandin-based estrus synchronization program from 14 to 7 days in cattle

Single injection of estrogen and progesterone before prostaglandin (steroid priming) was used to shorten the prostaglandin-based estrus synchronization program. Sixty-five cyclic Sistani cattle, with parity ranging from 1 to 4 and postpartum period of >80 days were selected at unknown stages of the estrous cycle and assigned to 2 groups according to their age, weight and parity. Females in the control group (n=33; 58.4 +/- 4.3 months; 277 +/- 8 kg LW) received two consecutive injections of prostaglandin F2alpha analogue (500 microg; Cloprostenol, PG) 14 days apart (Day 0 = First PG injection). On Day 7, treated females (n=32; 60 +/- 4.8 months; 292 +/- 9 kg LW) were given an intramuscular injection of 100 mg progesterone and 2 mg estradiol benzoate followed by prostaglandin 7 days later, concurrent with the second PG injection of the control group. Estrus detection was carried out every 6 hours for 7 days, commencing from 24 hours after the last PG injection. Females that allowed to be mounted were identified (standing estrus) and inseminated with frozen semen 12 hours later. Pregnancy was diagnosed on Day 50 after AI through palpation per rectum. Data were analyzed using Chi-squared and t-test. The tightness of estrus synchrony (%), the interval from the end of treatment to estrus (h) and conception rates (%) were similar (P > 0.05) between control (69.6%, 77.7 +/- 5.96 h and 56.5%) and treatment (68.2%, 82.6 +/- 7.64 h and 54.5%) groups. In conclusion, steroid priming is an efficient way to shorten the prostaglandin-based estrus synchronization program from 14 to 7 days without compromising estrous response and fertility.     

Effect of human chorionic gonadotropin administered at specific times following breeding on milk progesterone and pregnancy in cows

Two trials were conducted to investigate the efficiency of human chorionic gonadotropin (hCG) following breeding to increase progesterone (P(4)) secretion and pregnancy rates in cows. In Trial 1, 79 lactating Holstein cows were randomly allocated to four groups to receive hCG either at breeding (Day 0, n=20), Day 7 (n=20) or Day 14 (n=20), or to receive no hCG treatment (control n=19). Whole milk samples were collected every other day from breeding until Day 21 and, thereafter, at weekly intervals until Day 42 or until the return to estrus for determination of P(4) concentrations. Similar treatments were employed in Trial 2, and 121 lactating Holstein cows were randomly assigned to treatment at Day 0 (n=29), Day 7 (n=32), or Day 14 (n=29), or to receive no treatment and serve as a control group (n=31). Milk samples were obtained at weekly intervals from breeding until Day 42, or the return to estrus for determination of P(4) concentrations. Pregnancy diagnosis was made by palpation per rectum at approximately 60 days post breeding. Significant increases in P(4) concentrations were observed in Day-7 and Day-14 treated cows from Days 18 to 42 after breeding compared with the Day 0 or the control cows. A slight decrease in P(4) concentration throughout the sampling period was observed in the Day-0 treated cows. Significant increases in pregnancy rates were observed in hCG-treated cows compared with that of the controls, with the highest rate observed in the Day-7 treated group. The overall pregnancy rates were 47, 62, 55 and 40% for the Day 0, 7 and 14 groups and for the control groups, respectively. In nonpregnant cows the mean (+/- SEM) numbers of days to basal P(4) concentrations were 21.6 +/- 1.3, 24.1 +/- 1.6, 24.6 +/- 1.3 and 23.2 +/- 1.3 for cows treated on Days 0, 7 and 14 and for the control group, respectively. It is concluded that the administration of hCG at Day 7 or Day 14 after insemination could be used as a management tool to improve pregnancy rates in postpartum cows.     

Fertility in postpartum dairy cows in winter or summer following estrus synchronization and fixed time AI after the induction of an LH surge with GnRH or hCG

In this study, the fertility of postpartum dairy cows after a sequence of treatments with GnRH (Day 0), PGF2alpha (Day 7) and GnRH (Day 9) (GnRH group; n = 164) or hCG (Day 0), PGF2alpha (Day 7) and hCG (Day 9) (group hCG; n = 166) was investigated in summer and winter seasons. All cows were artificially inseminated without estrus detection, 16-18 h after the end of treatment. Control cows (CONT; n = 226) were not treated and were inseminated at natural estrus. The pregnancy rates at Day 90 (46% versus 33%; P < 0.05) and at Day 135 (76% versus 62%; P < 0.05) postpartum were significantly lower in CONT cows in summer compared to winter months but this effect was not observed in the two treated groups. The number of days from calving to conception was significantly lower in GnRH and hCG treatment groups compared to CONT cows in cold months (102 +/- 3.2, 106 +/- 4.2, 126 +/- 3.1, respectively; P < 0.001) and in hot months (112 +/- 3.2, 114 +/- 4.2, 139 +/- 3.1, respectively; P < 0.001). The concentration of insulin was significantly higher in winter (P < 0.001). There were no differences in average plasma concentration of glucose (P = 0.474), GH (P = 0.441) or IGF-I (P = 0.190). In conclusion, we have shown that veterinary supervision combined with a program of estrous synchronization and fixed time insemination can improve fertility of cows suffering heat stress.     

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 PGE2 on uterine contractility and tone in mares

A technique for transvaginal, ultrasound-guided intrauterine injection was developed. After preliminary study using different approaches, the procedure was successful in 24 of 25 (96%) mares, based on detecting fluid in the uterine lumen during and after the injection. The technique was used to study the effect of PGE2, reportedly produced by the embryonic vesicle, on uterine contractility on Day 12 (Day 0 = ovulation). Uterine contractility was scored (1 = minimal, 4 = maximal) every 10 min for 1 h and every 30 min for the next hour by a continuous 1-min ultrasound examination of a longitudinal section of the uterine body without knowledge of group. In Experiment 1, the main effect of group (1-mL vehicle, n = 6; 0.25 microgram PGE2, n = 7) tended to be significant (P < 0.09), and the effect of time was significant (P < 0.008). The mean score was higher for the PGE2 group (2.0 +/- 0.1) than for the vehicle group (1.7 +/- 0.1). An increase in contractility occurred between 0 and 5 min in the vehicle group (P < 0.0004) and between 0 and 10 min in the PGE2 group (P < 0.04). In Experiment 2, there was a tendency (P < 0.08) for effect of group (control without injection, n = 6; 1-mL vehicle, n = 6; 0.025 microgram PGE2, n = 6). The PGE2 group (2.0 +/- 0.1) was different from the vehicle group (1.6 +/- 0.1) and the control group (1.6 +/- 0.1). An increase in contractility occurred between 0 and 20 min in the PGE2 group, and the changes were not significant in the other groups. However, scores were higher in the PGE2 group before treatment, and there were no significant effects when data were converted to percentage changes. The results for an effect of intrauterine treatment of PGE2 on uterine contractility are considered uncertain because of the transient increase in contractility from vehicle injections in Experiment 1 and the higher score in the PGE2 group before treatment, with no significant differences in percentages in Experiment 2. Indirectly, however, an effect of PGE2 was suggested by a shorter (P < 0.05) period of detectability of intrauterine fluid in the PGE2 groups (21 +/- 31 min) than in the vehicle groups (50 +/- 42 min). The shorter period was attributable to greater dispersion of the fluid as a result of increased contractility. In Experiment 3, PGE2 (10 mg, n = 5) and vehicle (4 mL, n = 5) were given intravenously. In addition to uterine contractility, uterine tone was scored (1 = minimal, 4 = maximal) by transrectal digital compression. The main effect of group was significant (P < 0.03) for uterine contractility score, which increased between 0 and 20 min after PGE2 injection. The time effect and interaction were highly significant (P < 0.0001) for uterine tone score, and tone increased in the PGE2 group between 0 and 20 min after injection. The results indicated that PGE2 should be considered as a potential stimulator of both uterine contractions and uterine tone during the time of embryo mobility in mares.     

In vitro secretion of prostaglandins from endometrium of postpartum beef cows expected to have short or normal luteal phases

The objective of this study was to characterize endometrial secretion (in vitro) of prostaglandin F (PGF), 15-keto-13,14-dihydro-prostaglandin F2 alpha (PGFM), prostaglandin E2 (PGE2), and 6-keto-prostaglandin F1 alpha (6-keto-PGF1 alpha) on Day 5 following the first postpartum estrus of cows anticipated to have a short compared to a normal estrous cycle. Twenty-seven beef cows were randomly assigned into four groups. The Short Cycle (n = 6; control) and Short Cycle/Explant (n = 8; endometrial explants) groups had their calves weaned at 30-32 days postpartum. The Normal Cycle (n = 5, control) and Normal Cycle/Explant (n = 8; endometrial explants) groups received norgestomet (progestin) implants for 9 days beginning 21-23 days postpartum, and calves were weaned at implant insertion. Estrous cycle length (mean +/- SE; p less than 0.01) for the Short Cycle group was 11.5 +/- 1.9 days compared to 18.8 +/- 0.6 days for the Normal Cycle group. On Day 5 following the first postpartum estrus, cows in the Short Cycle/Explant and Normal Cycle/Explant groups were hysterectomized, and endometrial explants were incubated in Earle's Balanced Salt solution/Medium 199 for 90 min with or without arachidonic acid (AA) in the presence of three levels of oxytocin. Mean concentrations of PGF and PGFM were combined to obtain a value for total PGF. Concentrations of total PGF, PGE2 (from explants without AA treatment), and 6-keto-PGF1 alpha in medium of the Short Cycle/Explant group were higher (p less than 0.01) than in medium of the Normal Cycle/Explant group.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of intrauterine infusion of recombinant bovine interferon αI1 on luteal phase duration and oxytocin-induced release of 13,14-dihydro-15-keto-prostaglandin F2α in postpartum beef cows

The objective of this study was to determine if oxytocin-induced release of prostaglandin F₂α (PGF_2α; measured by the stable metabolite, 13,14-dihydro-15-keto-prostaglandin F₂α (PGFM)) was inhibited following intrauterine infusion of bovine interferon-α_I1 (rboIFNα_I1) into postpartum cows anticipated to have short estrous cycles following first ovulation postpartum. Cows expected to have short estrous cycles were assigned to receive twice daily intrauterine infusions of either placebo (SCP; n = 11) or 2 mg rboIFNα_I1 (SCIFN; n = 14) on Days 1–16 following hCG injection (2500 IU; day 0) on Days 30 or 31 postpartum. On Day 5 following hCG, each cow was injected with 100 IU oxytocin (i.v.) to induce the release of uterine PGF_2α (as measured by PGFM). Other treatment groups consisted of cows expected to have normal estrous cycle lengths following pretreatment with a 9 day norgestomet implant on Days 21 or 22 postpartum followed by hCG injection to induce ovulation. Cows expected to have normal estrous cycle lengths received twice daily intrauterine infusions of either placebo from Days 1 to 16 of the cycle and 100 IU oxytocin (i.v.) on Day 5 (NCPE; n = 11) or twice daily infusions of placebo (NCPL; n = 7) or rboIFNα_I1 (NCIFN; n = 10), from Day 13 post-hCG injection until luteolysis. Oxytocin was injected (100 IU; i.v.) into cows in the NCPL and NCIFN groups on Day 16. The calculated areas under the curve (arbitrary PGFM units) were: 164 ± 18 units, 96 ± 16 units, 93 ± 18 units, 137 ± 27 units and 53 ± 20 units for SCP, SCIFN, NCPE, NCPL and NCIFN, respectively (SCIFN < SCP; NCIFN < NCPL; P < 0.015). Mean luteal phase length was calculated as the number of days from injection of hCG until progesterone declined to below 0.5 ng ml⁻¹ and was: 6.7 ± 1.0 days, 10.5 ± 0.9 days, 12.0 ± 1.0 days, 18.0 ± 1.3 days and 20.7 ± 1.1 days for SCP, SCIFN, NCPE, NCPL and NCIFN, respectively (SCP < SCIFN = NCPE < NCPL = NCIFN; P < 0.01). In summary, luteal phase lengths were increased and oxytocin-induced release of PGFM was reduced by rboIFNα_I1 infusion in cows anticipated to have short luteal phases.     

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.     

Absorption of Escherichia coli endotoxin (lipopolysaccharide) from the uteri of postpartum dairy cows

An experiment was conducted to test the hypothesis that Escherichia coli (E. coli ) endotoxin is readily absorbed from uteri of early postpartum cows and that the absorbed endotoxin provokes systemic relcase of prostaglandins. Eleven postpartum Holstein dairy cows (aged 3 to 7 yr) with normal puerperium were selected and divided into a treatment group (n=7), which received intrauterine infusions of E. coli endotoxin, and a control group (n=4), which received intrauterine infusions of 10 ml of saline on Days 5 and 20 post partum. Blood samples were collected once every 30 min for 6 h starting from the time of infusion. Harvested sera samples were analyzed for concentrations of stable metabolites of prostacyclin (PCM), prostaglandin F(2alpha) (PGFM), and thromboxane A(2) (TXB(2)). Plasma samples were qualitatively tested for the presence of endotoxin. Endotoxin was detected in the plasma samples of cows that received endotoxin on Day 5 post partum 4 h after the infusion. Endotoxin was not detected in any of the samples from control cows on Days 5 and 20 post partum or from treatment group cows on Day 20 post partum. Cows treated on Day 5 post partum showed increases in serum PGFM concentrations from 710 +/-64pg/ml to peak concentrations of 1223 +/- 47 pg/ml within 2 h, followed by a decline to baseline concentrations within 4 h. The amount of PGFM released in treated cows on Day 5 post partum was higher (P < 0.05) than in control cows on Day 5 or in treated and control cows on Day 20 post partum. Serum PCM concentrations increased from 156+/-24 pg/ml to peak concentrations of 1348+/-127 pg/ml within 1 h. The amount of PCM released in treated cows on Day 5 postpartum was higher (P< 0.05) than in control cows on Day 5 or in treated and control cows on Day 20 post partum. The TXB(2) concentrations increased from 315+/-38 pg/ml to peak concentrations of 5043 +/- 242 pg/ml within 1 h and fell to baseline concentrations within 5 h. The amount of TXB(2) concentrations released in treated cows on Day 5 post partum was significant (P < 0.05) compared with those of cows in the other groups. The results support the hypothesis that uteri of early postpartum cows are capable of absorbing endotoxin, and the absorbed endotoxin provokes changes in the serum concentrations of prostanoids.     

Ovulation and follicular growth in gonadotropin-treated gilts followed by in vitro fertilization and development of their oocytes

We investigated whether porcine ovaries derived from FSH-pituitary (FSH-P) or hCG-treated animals can produce oocytes with better in vitro cytoplasmic maturation and in vitro embryonic development relative to those derived from saline-treated animals. The size of the follicle producing the oocyte was also studied. Each of 25 prepubertal gilts received 1 of 6 treatments by intramuscular injection: 1) saline (3 mL, once, n = 5); 2) FSH-P8-3 (8 mg, 3 times, with a 24-h interval, n = 4); 3) FSH-P16-3 (16 mg, 3 times, with a 24-h interval, n = 4); 4) FSH-P16-1-P4-2 (16 mg, once, 4 mg, twice, with a 24-h interval, n = 4); 5) FSH-P16-1 (16 mg, once, n = 4); or 6) hCG (100 IU, 3 times, with a 24-h interval, n = 4). The ovaries were removed by mid-ventral laparotomy 72 h after the first injection. The numbers of corpora hemorrhagica (CH) with each FSH-P treatment were similar (P > 0.05). However, compared with gilts treated with saline or hCG, those treated with FSH-P8-3 had a greater (P < 0.05) number of CH. Treatment with FSH-P8-3 or FSH-P16-3 induced significant growth of medium/large follicles (4 to 8 mm in diameter) compared with saline or FSH-P16-1. The same results were observed when FSH-P8-3 was compared with FSH-P16-P4-2 or hCG. After in vitro fertilization, the rates of male and female pronuclei in oocytes derived from medium/large follicles did not differ (P > 0.05) between treatments, but in oocytes derived from small follicles they were lower (P < 0.05) in saline-treated than in FSH-P16-1-P4-2-treated gilts. After 120 h in culture, the percentages of the inseminated oocytes from 1 to 3 mm or 4 to 8 mm follicles developing to > or = 2-cell did not differ (P > 0.05) between saline- and gonadotropin-treated gilts. However, a higher (P < 0.05) percentage of the inseminated oocytes from 4 to 8 mm follicles had developed to the morula stage or beyond, than those from the 1 to 3 mm follicles. In conclusion, administration of single or multiple doses of FSH-P induced ovulation, but only 8 or 16 mg FSH-P injected 3 times with 24-h intervals for 72 h induced growth of 4 to 8 mm follicles. The size of follicle from which the oocyte derived also had a significant effect on its development in vitro.