Plasma steroid profiles following follicle-stimulating hormone or equine chorionic gonadotropin injection in cows chronically treated with gonadotropin-releasing hormone agonist

Plasma steroid profiles following follicle-stimulating hormone (FSH) or equine chorionic gonadotropin (eCG) injection were studied in chronically gonadotropin releasing hormone agonist (GnRH-A)-treated cows. Follicular development and irINH secretion were stimulated by FSH or eCG injection. The plasma concentrations of estradiol-17 beta (E(2)) and testosterone (T) were markedly increased following eCG injection. However, significant increases of the plasma E(2) and T concentrations were not detected in FSH-treated cows. Ovulation of developed follicles were depended on the hCG injection in both groups. These results show: 1) Follicular response to an exogenous gonadotropin is still remained, 2) Ovulation of developed follicles is induced by hCG injection and 3) FSH and eCG cause disparate plasma steroid profiles, under the influence of repeated GnRH-A treatment.     

Treatment of post-partum anoestrous dairy cows with progesterone, oestradiol and equine chorionic gonadotropin

Anoestrus in lactating dairy cows at the start of the breeding season is a major form of reproductive wastage for seasonal dairy production based on pasture. The objective of this study was to compare the reproductive performance of anoestrous cows that were treated with a combination of progesterone, oestradiol and equine chorionic gonadotrophin either 10 days before (T-10, n = 2 19) or 16 days after (T+16, n = 229) the start of the breeding season. A higher percentage of cows in the T-10 group were detected in oestrus and inseminated during the first 6 days of breeding than those in the T+16 group (69.4% v. 26.2%. p <0.001). However, the percentage of cows detected in oestrus by Day 16 was similar between the two treatment groups (T10 v. T+ 16; 77.7% v. 76.7%). There was no difference between treatment groups in conception rate to the first (51.2% v. 59.0%) or the second insemination (50.8% v. 57.6%), in pregnancy rate over the first 49 days (74.0% v. 75.1%), in empty rate (10.0% v. 10.5%) or in the mean day of conception from the start of the breeding season (24.0 v. 25.7 days). These results suggest that, under favourable environmental conditions, treatment of anoestrous cows with the programme used in this trial can be performed 16 days after the start of the breeding season with similar results to that performed 10 days before the start of the breeding season. Further studies are needed to determine if this is the case under different environmental conditions or for other treatment programmes.     

Administration of human chorionic gonadotropin to suckled beef cows before ovulation synchronization and fixed-time insemination: replacement of gonadotropin-releasing hormone with human chorionic gonadotropin

Two experiments were conducted to evaluate whether hCG administered 7 d before initiating the CO-Synch + controlled internal drug release (CIDR) ovulation synchronization protocol (Exp. 1 and 2), or replacing GnRH with hCG at the time of AI (Exp. 1), would improve fertility to a fixed-time AI (TAI) in suckled beef cows. In addition, the effects of hCG on follicle dynamics, corpus luteum development, and concentrations of progesterone (P4) were evaluated. In Exp. 1, cows were stratified by days postpartum, age, and parity and assigned randomly to a 2 × 2 factorial arrangement of 4 treatments: 1) cows received 100 μg of GnRH at CIDR insertion (d -7) and 25 mg of PGF(2α) at CIDR removal (d 0), followed in 64 to 68 h by a TAI plus a second injection of GnRH at TAI (CG; n = 29); 2) same as CG but the second injection of GnRH at the time of insemination was replaced by hCG (CH; n = 28); 3) same as CG, but cows received hCG 7 d (d -14) before CIDR insertion (HG; n = 28); and 4) same as HG, but cows received hCG 7 d (d -14) before CIDR insertion (HH; n = 29). Pregnancy rates were 52, 41, 59, and 38% for GG, GH, HG, and HH, respectively. Cows receiving hCG (39%) in place of GnRH at TAI tended (P = 0.06) to have poorer pregnancy rates than those receiving GnRH (56%). Pre-CO-Synch hCG treatment increased (P < 0.05) the percentage of cows with concentrations of P4 >1 ng/mL at d -7, increased (P < 0.02) concentration of P4 on d -7, and decreased (P < 0.001) the size of the dominant follicle on d 0 and 3, compared with cows not treated with hCG on d -14. In Exp. 2, cows were stratified based on days postpartum, BCS, breed type, and calf sex and then assigned to the CG (n = 102) or HG (n = 103) treatments. Overall pregnancy rates were 51%, but no differences in pregnancy rates were detected between treatments. Pre-CO-Synch hCG treatment increased (P < 0.05) the percentage of cows cycling on d -7 and increased (P < 0.05) concentrations of P4 on d -7 compared with pre-CO-Synch controls. Therefore, pretreatment induction of ovulation after hCG injection 7 d before initiation of CO-Synch + CIDR protocol failed to enhance pregnancy rates, but replacing GnRH with hCG at the time of AI may reduce pregnancy rates.     

Biological activities of tethered equine chorionic gonadotropin (eCG) and its deglycosylated mutants

Equine chorionic gonadotropin (eCG), which consists of highly glycosylated alpha- and beta-subunits, is a unique member of the gonadotropin family because it elicits response characteristics of both follicle-stimulating hormone (FSH) and luteinizing hormone (LH) in species other than the horse. In this study, recombinant tethered-eCG as well as its deglycosylated mutants were produced to determine if alpha- and beta- subunits can be synthesized as a single polypeptide chain (tethered-eCG) and display biological activity. We found that tethered-eCG (T- betaalpha) had both LH- and FSH-like activities comparable to dimeric eCG. Luteinizing hormone-like activity of tethered-eCGs deglycosylated at Asn(56) (T-betaalpha56) was decreased. In contrast, LH-like activity of eCG without O-glycosylated carboxyl-terminal peptide (CTP) (T-betacalpha) was slightly decreased but still similar to T-betaalpha. Double mutation at Asn(56) and CTP (T-betacalpha56) caused marked decrease in the activity, indicating that both glycosylations at Asn(56) and CTP are involved in LH-like activity in the tethered form. Interestingly, FSH-like activity remained in all deglycosylated eCG mutants (T-betaalpha56, T-betacalpha and T-betacalpha56) as well as T-betaalpha. The biological roles of oligosaccharides at Asn(56) of eCG alpha-subunit and O-linked peptide of beta-subunit appear to be different in LH- and FSH-like activities in tethered-eCG.     

Ovulation in postpartum beef cows treated with estradiol

The effect of implants of estradiol on initiation of ovarian cycles postpartum was studied in 201 anestrous beef cows. Cows from four farms were used over a 2-yr period in a 2 x 3 factorial arrangement of treatments with estradiol implants and stage postpartum as main effects. Cows were assigned at random within date of calving within farm to receive an ear implant containing estradiol-17 beta (24 mg) for 21 d or to serve as controls. Stages postpartum at implantation were divided into less than or equal to 25, 26 to 39, and greater than or equal to 40 d, three stages that should reflect potential changes in hypothalmic-hypophysial sensitivity to estradiol. Blood samples for determination of progesterone were obtained and rectal examinations of the ovaries performed at implant insertion, 14 d after insertion, at implant removal (d 21), and 14 d after removal (d 35) to assess ovulatory response to treatment. Circulating concentrations of estradiol on d 14 of treatment averaged 3.2 +/- 1.0 and 23.1 +/- 4.7 pg/ml for control and estradiol-treated cows, respectively. Compared with control cows, treatment with estradiol initiated after d 26 postpartum increased the proportion of cows that ovulated during the experimental period. No differences were seen in the average days postpartum when cows were first determined to have ovulated.     

Follicular phase gonadotropin secretion in cyclic postpartum beef cows with phlorizin-induced hypoglycemia

Twenty mature, lactating Hereford-cross cows were used to determine the effect of phlorizin-induced hypoglycemia on gonadotropin secretion following prostaglandin-induced luteolysis. Cows were 43 to 108 d postpartum and had a functional corpus luteum (CL) at the start of infusion treatment (d 1). Infusions consisted of either saline (control) or 3 g/d of phlorizin infused continuously from the time of prostaglandin injection at 1000 on d 1 until 0800 on d 5. Blood samples were collected for determination of plasma concentrations of insulin, glucose and free fatty acids (FFA) and for serum concentrations of luteinizing hormone (LH), follicle-stimulating hormone (FSH) and progesterone. Plasma concentrations of insulin (P less than .05) and glucose (P less than .05) were lower, whereas FFA concentrations increased (day X treatment, P less than .05) over the infusion period in phlorizin-treated cows compared with saline-infused controls. Mean serum concentrations of LH (1.17 +/- .10 vs 1.53 +/- .20 ng/ml; P less than .05) and LH pulse amplitude (1.69 +/- .14 vs 2.47 +/- .37 ng/ml; P less than .10) were lower in phlorizin-infused compared with saline-infused cows during the 0 to 24-h period immediately preceding the ovulatory gonadotropin surge. The FSH pulse frequency increased (.33 +/- .11 to .55 +/- .12 pulses/h) in saline-infused cows, but decreased (.61 +/- .10 to .41 +/- .11 pulses/h) in phlorizin-infused cows before the gonadotropin surge. Other characteristics of gonadotropin secretion were similar among phlorizin-infused and saline-infused cows. All but one phlorizin-infused cow ovulated and formed functional CL similar to controls.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of equine chorionic gonadotropin on reproductive performance in anestrous mink.

The incidence of anestrous mink during the normal breeding season has been reported to be as high as 5%. We sought to induce estrus in these mink by using various doses of equine chorionic gonadotropin (eCG) and human chorionic gonadotropin (hCG). Seventy-five female mink maintained under standard ranch conditions failed to demonstrate estrus during the annual breeding season in March. These anestrous mink were randomly assigned to treatment groups. Treatments were given on March 16 and again on March 18. On these 2 d, mink were treated with equal doses of saline, 25, 50, or 100 IU of eCG, or 50 IU of eCG on March 16 and 50 of IU of eCG + 100 IU of hCG on March 18. Females were paired with males beginning on March 22. None of the saline-treated mink mated. In contrast, reproductive performance of the anestrous mink was significantly improved by treatment with eCG. This included proportion of mink breeding (47 to 100%), proportion giving birth (33 to 80%), and average litter size (2.6 to 4.0 kits per whelping female). Reproductive efficiency improved with increasing doses of eCG and was not further improved by addition of hCG. These results demonstrate that eCG has a potential application for treating anestrous mink during the breeding season.     

Effect of morphine on serum gonadotropin concentrations in postpartum beef cows

Morphine (M), an opioid agonist, was administered to postpartum (PP) Angus cows to investigate opioid modulation of gonadotropin secretion. In Exp. 1, eight PP cows (36.9 +/- 2.3 d) received either M (1 mg/kg; n = 4) or saline solution (S) (n = 4) via i.v. injection 36 h after calf removal. Morphine decreased (P less than .01) the number of serum LH pulses (3.0 +/- 1.1 pre- vs .3 +/- .3 post-pulses/h) and, compared with pretreatment values (3.3 mg/ml), decreased (P less than .05) mean LH at 105 min (2.1 ng/ml) through 270 min 1.9 ng/ml +/- .4). Serum prolactin (PRL) increased (P less than .01) following M from 16.4 ng/ml to a peak of 59.3 ng/ml (+/- 3.9). Serum FSH concentrations were unaffected. In Exp. 2, M (.31 mg/kg i.v. injection followed by .15 mg/(kg.h) infusion; n = 6) or S (n = 6) treatments were given for 7 h beginning 36 h after calf removal. Serum LH was similar between groups during the pretreatment and the first 6 h of infusion, but M decreased (P less than .001) the number of serum LH pulses (.44 +/- .09 vs .06 +/- .04 pulses/h). Morphine increased (P less than .05) serum PRL. It is concluded that M differentially modulated gonadotropin secretion in the cow such that PRL increased, LH decreased and FSH was unchanged.     

Induction of oestrus by administering Inhibin antiserum along with equine chorionic gonadotropin in anoestrous bitches

As dogs experience oestrus only once or twice a year, it is necessary to establish an effective method of oestrous induction for efficient breeding. In the present study, we evaluated inhibin antiserum (IAS) on oestrous induction in anoestrous females. Bitches were administered 0.5 ml/kg IAS or a mixture of 50 IU/kg equine chorionic gonadotropin (eCG) and 0.5 ml/kg IAS and 500 IU human chorionic gonadotropin (hCG) administered 7 days after the mixture injection. As a control, bitches received 50 IU/kg eCG, with 500 IU hCG administered 7 days after eCG injection. Blood-tinged vaginal discharge, vulvar swelling, plasma progesterone concentrations and ovarian follicular development were assessed from day 0 to day 14. IAS alone injection did not induce oestrus in bitches at the anoestrous stage. Conversely, vulvar swelling, blood-tinged vaginal discharge and an estimated luteinizing hormone (LH) surge appeared on days 3–7, days 3–6 and days 7–9 after the IAS+eCG mixture injection, respectively, in all five bitches at the anoestrous stage. The average number of developing and ovulated follicles in bitches administered IAS+eCG was 8.8 and 9.6 respectively. A single eCG injection followed by hCG induced oestrous signs, with an average of 8.3 developing follicles and 4.5 ovulated follicles. This study revealed that IAS alone did not induce oestrus, but when IAS was used in combination with eCG, it induced oestrus and promoted a considerable number of ovulations in anoestrous dogs.     

Response of postpartum beef cows to exogenous progestogens and gonadotropin releasing hormone

Plasma progesterone (P4) profile and estrous detection were used during three experiments to evaluate the effects of exogenous progestogens on the life span of gonadotropin releasing hormone (GnRH)-induced corpora lutea (CL) in postpartum (pp) beef cows. Experiment 1 utilized primiparous fall-calving cows (n = 28, trial 1); and spring-calving cows (n = 29, trial 2). On d 18 to 27 pp (d 0) all cows received intravaginal devices containing either P4 or no P4 (NP) for 5 d. On d 5 the devices were removed and calves were either removed (CR) or were present (CP) with half of the cows within steroid group. At 50 h after device removal, 500 micrograms of GnRH was given (iv) to all cows, and weaned calves were reunited with their dams. The induced CL had a normal life span (greater than 16 d) in 17 and 86% (trial 1) and 8 and 79% (trial 2) of NP and P4 cows, respectively. Calf removal did not affect (P greater than .10) the life span of the CL. In Exp. 2, spring-calving multiparous cows (d 18 to 24 pp; d 0) received either no P4 (NP; n = 19), P4 for 6 d via intravaginal devices (P4H; n = 19) or a single im injection of 300 mg P4 (P4 IM; n = 18). At 48 h after device removal or at 8 d after the injection of P4, half of the cows within steroid group received either 500 micrograms GnRH or saline. Corpora lutea had a normal life span in 0, 11, and 80% of NP, P4 IM and P4H cows, respectively, that received GnRH and in 22% of P4-saline cows. In Exp. 3, fall-calving multiparous and primiparous cows (d 25 to 31 pp) received either no progestogen (NP; n = 20), P4 via intravaginal devices for 5 d (P4H; n = 21) or melengestrol acetate (MGA; .5 mg.head-1.d-1 for 5 d orally, n = 15). At 48 d after device removal or at 72 h after the last MGA feeding, all cows received 500 micrograms GnRH. Progesterone post-GnRH injection was increased (greater than 1 ng/ml) at d 7 in 64, 100 and 100%, and remained elevated at d 14 in 11, 46 and 100% of NP, MGA and P4H cows, respectively. For all experiments plasma P4 was increased (range 2 to 5 ng/ml) when the devices containing P4 were in place, then decreased (less than 1 ng/ml) by 48 to 50 h after device removal.(ABSTRACT TRUNCATED AT 400 WORDS)     

Steroids and cAMP in follicles of postpartum beef cows treated with norgestomet.

To determine time of occurrence of follicular changes that may be associated with the length of the subsequent luteal phase, follicles were collected at different times before ovulation from cows expected to have corpora lutea of short (control) or normal (norgestomet-treated) life span. Beginning on d 20 to 23 postpartum (d 0 of study), 34 crossbred beef cows received either a 6-mg implant of norgestomet for 9 d or served as untreated controls. Ovaries were removed from norgestomet-treated cows on d 6 (N6; n = 9), d 8 (N8; n = 8), or the day after implant removal (N10; n = 8). Control cows were ovariectomized on d 6 (C6; n = 4) or d 10 (C10; n = 5). The largest and second largest follicles greater than 8 mm (F1 and F2, respectively) were dissected from the ovaries. Granulosal and thecal layers and follicular fluid were separated and assayed for estradiol-17 beta, progesterone, androstenedione, and testosterone. Cyclic 3'5'adenosine monophosphate (cAMP) was determined in thecal and granulosal tissue. Diameters of the F1 (14.6 +/- .4 mm) and F2 (10.6 +/- .4 mm) did not differ due to treatment. A greater proportion (P less than .05) of the F1 (20/33) than of the F2 (4/27) had estradiol:progesterone ratios of greater than 1 in follicular fluid. Contents of estradiol, androstenedione, and testosterone in theca and granulosa and follicular fluid, androstenedione in theca, and testosterone in theca and follicular fluid (all P less than .05).(ABSTRACT TRUNCATED AT 250 WORDS)     

Treatment of suckling beef cattle with a progestagen sponge and oestradiol benzoate or equine chorionic gonadotrophin

The ovarian responses of anoestrus beef cows to a combined treatment with medroxy-progesterone acetate (MAP) sponges and oestradiol benzoate or equine chorionic gonadotrophin (eCG) were evaluated. Forty-five suckling Hereford cows were allocated to three equal groups. Group 1 received a MAP sponge for seven days plus an injection of 2 mg oestradiol benzoate when the sponge was inserted (day 0) and 1 mg when the sponge was withdrawn; group 2 received identical treatment until day 7, when a dose of 400 iu of eCG was administered, and group 3 were left untreated as control animals. From day 0 to day 11 the cows' ovaries were examined daily by transrectal ultrasonography, and their oestrous behaviour was observed from 24 hours to 96 hours after the sponge was removed. Data from cows that had a corpus luteum present before the sponge was withdrawn were not used in subsequent analyses; there were four in group 1, five in group 2 and four in group 3. In 19 of the 21 cows in groups 1 and 2 a new follicular wave was observed to emerge at a mean (sd) interval of 3.9 (0.3) days after the insertion of the sponge, whereas in group 3 it occurred in all 11 cows after 3.4 (0.6) days. Only the six cows that had a follicle of 9 mm or larger in diameter ovulated (P < or = 0.001). Nine of the 11 cows in group 1 came into oestrus, compared with two of the 10 in group 2 and none of the control cows (P < or = 0.001). Ovulation was observed in four, two and none of the cows in groups 1, 2 and 3, respectively.     

Fixed-time artificial insemination in replacement beef heifers after estrus synchronization with human chorionic gonadotropin or gonadotropin-releasing hormone

We determined the effects of hCG on ovarian response, concentration of progesterone, and fertility in a fixed-time AI (TAI) protocol. Four hundred forty-four crossbred beef heifers were synchronized with the CO-Synch + CIDR (controlled internal drug-releasing insert) protocol. In addition, heifers were randomly assigned to 1 of 4 treatments in a 2 × 2 factorial arrangement of treatments with main factors being 1) pretreatment, no treatment (control), or treatment with 1,000 IU of hCG 14 d before the initiation of the CO-Synch + CIDR protocol and 2) treatment, administration of 1,000 IU of hCG or 100 μg of GnRH at CIDR insertion of the CO-Synch + CIDR protocol. Blood samples were collected from all heifers on d -21, -14, -7, 0, and 2 relative to PGF(2α) injection. Transrectal ultrasonography was used to examine ovaries in a subset of heifers (n = 362) on d -7 and 0 relative to PGF(2α), and to determine pregnancy status of all heifers on d 33 and 82 relative to AI. Pregnancy rates were similar for heifers pretreated with control (33.0%) or hCG (36.4%), whereas pregnancy rates were greater (P < 0.01) for heifers treated with GnRH (40.1%) compared with hCG (29.0%) at CIDR insertion. Heifers pretreated with hCG had more (P < 0.01) corpora lutea present on the day of CIDR insertion and the day of CIDR removal compared with untreated heifers. A greater proportion (P < 0.01) of heifers ovulated as a result of administration of hCG at the time of CIDR insertion (59.0%) compared with GnRH (38.7%). Heifers treated with hCG at CIDR insertion had greater (P < 0.01) concentrations of progesterone compared with those receiving GnRH at the time of CIDR removal (2.42 ± 0.13 vs. 1.74 ± 0.13 ng/mL; P < 0.01) and at fixed-time AI (0.52 ± 0.03 vs. 0.39 ± 0.03 ng/mL; P < 0.01). Therefore, hCG was more effective than GnRH in its ability to ovulate follicles and to increase concentrations of progesterone in beef heifers. Presynchronization with hCG 14 d before CIDR insertion did not alter pregnancy rates, whereas replacing GnRH with hCG at CIDR insertion decreased pregnancy rates.     

Reproductive performance of sows treated with a combination of pregnant mare's serum gonadotropin and human chorionic gonadotropin at weaning in the summer

During the summer and fall of 1987, sows from eight herds in three states were assigned randomly to receive either a combination of 400 IU of pregnant mare's serum gonadotropin with 200 IU of hCG (P.G. 600) or no treatment at weaning. A treatment x parity interaction was observed for days to first estrus after treatment and percentage anestrus (percentage of sows not achieving estrus within 10 d after weaning). Relative to primiparous control sows, primiparous sows given P.G. 600 expressed estrus sooner (P less than .02) after weaning (6.0 vs 7.8 +/- .6 d) and exhibited less (P less than .02) postweaning anestrus (15.6 vs 29.2 +/- 4.0%). Second parity sows that received P.G. 600 showed estrus sooner (P less than .06) than second-parity control sows (4.7 vs 6.4 +/- .7 d). Days to first estrus after treatment did not differ between groups for parity-three and older sows, and percentage anestrus was not different between treatments for parity-two and older sows. The herd X treatment interaction was significant for percentage recycled (percentage of successfully mated sows that returned to estrus), subsequent farrowing rate, and subsequent number of pigs born dead. Number of pigs born alive was lower for sows treated with P.G. 600 than for control sows (10.55 vs 10.10 +/- .18; P less than .02). In summary, treatment of sows weaned in the summer and fall with P.G. 600 had decreased days to postweaning estrus in parity-one and -two sows and reduced frequency of postweaning anestrus in primiparous sows.     

Evaluation of melengestrol acetate and equine chorionic gonadotropin for out-of-season breeding in sheep on Prince Edward Island

The purpose of this study was to compare the efficacy of a recommended protocol of oral melengestrol acetate (MGA) to intravaginal medroxyprogesterone acetate (MPA), with or without equine chorionic gonadotropin (eCG), for out-of-season breeding of sheep on Prince Edward Island. One hundred and twenty ewes were assigned to 1 of 4 groups in a factorial design and were treated with either an intravaginal MPA sponge or oral MGA. Ewes received either an intramuscular injection of eCG or a saline placebo at cessation of progestogen treatment. The reproductive performance was best for the MPA-eCG group, followed by the MGA-eCG, MGA-saline, and MPA-saline. Pregnancy rates of 66.7%, 43.3%, 31.0%, and 16.7%, respectively, and ratios of lambs born to ewes exposed to the ram of 1.17, 0.63, 0.52, and 0.23, respectively, were recorded for these groups. We concluded that, under the conditions of this study, the use of oral MGA resulted in fewer lambs than did the use of MPA sponges with eCG. Nonetheless, the use of MGA may be attractive to producers because it is less expensive and more convenient than the use of sponges.