Effects of the dam on equine chorionic gonadotropin concentrations during pregnancy

Breeding trials were designed to determine the influence of the mare on serum concentrations of equine chorionic gonadotropin (eCG) from Day 39 to Day 104 of gestation. Sires were ranked according to mean eCG concentrations found in the groups of randomly selected mares to which they were mated in 1983. Mares were ranked according to their mean eCG concentrations on Days 55, 71 and 85 of gestation (Day 0 = mating), in 1983 and 1985. In the 1986 breeding season, mares that had pregnancies characterized by high eCG levels were mated to sires previously associated with low eCG concentration pregnancies and low producing mares were mated to sires associated with high eCG concentration pregnancies. The highest eCG concentrations (12.8 ug/ml serum) were detected on Day 55 of gestation in mares mated to the low-ranked sire (P<0.01), indicating an influence of the mare on serum eCG concentration. A comparison of eCG production by individual mares in 1985, 1986 and 1987 showed that mares retained the same rank (P<0.001), regardless of the rank of the sire to which they were mated. It was concluded that the influence of the mare predominates in determining eCG concentration. In 1987, the highest ranked sire was mated to the highest eCG producing mares (ranked 1–20) and the second highest ranked sire was mated to mares ranked 21–40. Two distinct subgroups resulted, suggesting that the selection of sires and mares could be used to maximize the production of eCG.     

Changes in plasma progesterone concentrations in mares treated with cloprostenol and human chorionic gonadotropin and inseminated during estrus

Daily changes in the plasma progesterone concentrations were determined in eight mares treated with intramuscular injections of 250 μg cloprostenol, a prostaglandin analogue, followed five days later by 2500 I.U. human chorionic gonadotropin. A second cloprostenol injection was given 14 days after the first; the mares were then inseminated on the third and fifth day of the subsequent estrus and a second injection of human chorionic gonadotropin was administered on the fifth day. The onset of estrus following the second cloprostenol treatment was synchronized beginning three to four days after treatment in all eight mares. All eight ovulated, five mares conceived and only four foaled. Evaluation of the progesterone profiles provided reliable indicators of luteolysis, ovulation and luteal function. Decreasing plasma progesterone concentrations were associated with cloprostenol induced luteolysis or preceded spontaneous onset of estrus. The plasma progesterone concentrations increased consistently after ovulation, and in the pregnant mares, the progesterone concentrations remained high during the first month after insemination.     

Effects of a single injection of GnRH or equine pituitary extract on plasma LH and FSH concentrations in stallions

Plasma LH and FSH concentrations were measured in mature stallions after administration of synthetic GnRH or equine pituitary extract. GnRH caused significant rises in plasma LH (2-fold) and FSH (1.7-fold). Concentrations of LH remained significantly elevated for 4 hours and FSH remained elevated for 2 hours. Similar increases in plasma LH (1.6-fold) and FSH (1.8-fold) occurred after an injection of equine pituitary extract. LH was significantly elevated for 4 hours and FSH was elevated for 6 hours.     

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.     

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.     

Pregnancy and twinning rates in Thoroughbred mares following the administration of human chorionic gonadotropin (hCG)

AIM: To determine the effect of hCG administration to cycling Thoroughbred mares, on pregnancy and twinning rates and the number of serves in the treated cycle. METHODS: A retrospective case control approach was conducted involving 2119 mare ovulatory cycles, on 1110 mares over a 7-year period. Data were collected by 1 of the authors during routine stud work at 3 commercial Thoroughbred farms in the Waikato region of New Zealand. The hCG (1500 IU) was administered by intravenous injection to selected mares 24 h before the expected time of breeding. Mares were scanned for pregnancy (singleton or twins) 14 days after the onset of dioestrus or detection of ovulation. Multilevel logistic regression analyses were used to identify the risk factors associated with the outcomes of interest while simultaneously controlling for possible confounding factors. RESULTS: Treatment with hCG tended to improve the odds of pregnancy (p=0.06), produced a 3-fold increase in the odds of twins (p<0.001), and increased the odds of a mare having a single serve in the treated ovulatory cycle (p=0.036). The first ovulatory cycle of a season in which a mare was bred was associated with a lower odds of pregnancy (p=0.02), and a lower odds of twins (p=0.003), when compared with subsequent cycles. Lactating mares were less likely to be diagnosed with twins (p=0.005), and were more likely to have a single serve (p<0.001), in any one ovulatory cycle than non-lactating mares. CONCLUSIONS: This report supports the role of hCG as an important therapeutic tool in veterinary management of broodmares for optimal reproductive performance. Mares treated with hCG must be managed in the knowledge that they have an increased likelihood of twins.     

Hormone therapy for control of reproduction in mares and stallions

Because the reproductive performance of mares is lower than that of any other domesticated species, hormone therapy is important in ensuring fertility and proper management of pregnancy. Current techniques of hormone therapy are discussed.     

The pregnancy protecting effect of progesterone against human chorionic gonadotrophin challenge in mares

14 pregnant Welsh Mountain Pony mares were treated with progesterone in an effort to prevent pregnancy failure induced by administration of human chorionic gonadotropin (GCG). 13 of the 14 mares were treated with progesterone by injection or implant before the 38th day of pregnancy. HCG was given in 3 doses on alternate days at a dose rate of 2000 imc/day. The remaining mare was treated with HCG toward the end of the experiments to demonstrate the abortifacient property of HCG. 3 mares aborted and 1 of these was anovulatory thereafter. Further research is needed to determine the effective progesterone dose and administration regime for mares thought to be suffering from insufficient luteal activity during early pregnancy.     

Human chorionic gonadotropin (hCG): the effect of dose on ovulation and pregnancy rate in Thoroughbred mares experiencing their first ovulation of the breeding season

AIM: To determine the effect of hCG dose on ovulation and pregnancy rate in Thoroughbred mares experiencing their first ovulation of the breeding season. METHODS: Over 3 successive breeding seasons, a total of 101 mares were randomly assigned to 1 of 4 treatment groups (intravenous injection of either saline, 1500, 3000, or 6000 IU hCG), as they approached their first ovulation of the breeding season. Mares were bred 1 day post-injection to 1 of 11 stallions, and every other day until ovulation occurred. Data were analysed using multivariable logistic regression with correction for over-dispersion due to clustering. RESULTS: Mares treated with hCG were more likely to ovulate within 72 h of treatment than mares treated with saline (p<0.001); there was no significant difference between doses of hCG on risk of ovulation (p>0.15). Farm also had a significant impact on the risk of ovulation (p=0.027). Mares treated with hCG were more likely to be diagnosed pregnant 14 days post ovulation than saline-treated mares (p=0.081, p=0.029 and p=0.026 for the 1500, 3000 and 6000 IU doses, respectively); there was no significant difference between doses of hCG on risk of pregnancy (p>0.45). CONCLUSIONS: A single injection of hCG (1500-6000 IU) is effective at inducing ovulation in late transitional mares and increases the likelihood of pregnancy at 14 days post ovulation. This paper supports the use of hCG as an integral part of optimal broodmare management.     

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.     

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.     

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.     

Intervals from norgestomet withdrawal and injection of equine chorionic gonadotropin or P.G. 600 to estrus and ovulation in ewes

Synchronization of estrus and ovulation is essential for AI of ewes during a predetermined time frame, and progestogen-eCG treatments are typically used to prepare the ewes. However, eCG is not readily available in the United States, but P.G. 600 (400 IU of eCG and 200 IU of hCG) is available. Thus, we conducted a study to determine the effects of eCG and P.G. 600 on the timing of estrus and ovulation after progestogen withdrawal. Ewes were assigned to two replicates of an experiment with the following treatments: 1) 3-mg norgestomet implant (i.e., one-half of a Syncro-Mate-B [SMB] implant) for 10 d, plus 2 mL of saline i.m. at SMB removal (n = 11); 2) 3-mg SMB implant for 10 d, plus 400 IU of eCG i.m. at SMB removal (n = 13); and 3) 3-mg SMB implant for 10 d, plus P.G. 600 i.m. at implant removal (n = 9). On d 6 after SMB insertion, PGF2alpha was used to induce luteolysis. Beginning 12 h after implant removal, vasectomized rams were used at 12-h intervals to check for estrus. When a ewe was detected in estrus, each ovary was evaluated ultrasonically. Ovaries were evaluated again 16 h later and then at 8-h intervals until ovulation. Treatment altered the interval from implant removal to estrus (less [P < 0.05] in SMB + eCG than in the other two groups) and to ovulation (greatest [P < 0.05] in SMB). However, the treatment x replicate interaction was significant for the intervals from implant removal to estrus (P < 0.03) and from implant removal to ovulation (P < 0.05). An inconsistent response in the SMB-treated ewes seemed to be primarily responsible for the interaction. The intervals to estrus and to ovulation for the SMB-treated ewes were shorter (P < 0.05) in Replicate 1 than in Replicate 2. Also, both intervals seemed to be less consistent between replicates for the SMB + P.G. 600- than for the SMB + eCG-treated ewes; that is, eCG seemed to increase the predictability of the intervals to estrus and to ovulation. Neither the main effects of treatment and replicate nor their interaction were significant for the interval from estrus to ovulation (38.4 /- 3.3 h), size of the ovulatory follicle (7.7 +/- 0.8 mm), or ovulation rate (1.6 +/- 0.2). We concluded from this experiment that eCG is a better choice than P.G. 600 as the gonadotropin to use at the time of progestogen withdrawal to prepare ewes for AI during a predetermined interval.     

Changes of plasma concentrations of steroid hormones, prostaglandin F2 alpha-metabolite and pregnant mare serum gonadotropin during pregnancy in thoroughbred mares.

Plasma concentrations of estrogens, gestagens, cortisol (F), 13, 14-dihydro, 15-keto PGF2 alpha (PGFM) and pregnant mare serum gonadotropin (PMSG) in 10 Thoroughbred mares were measured for a 11-month pregnancy period. Estrone (E1) and estradiol-17 beta (E2) levels gradually increased as the pregnancy advanced, showing a peak around Month 8 and decreased thereafter. Progesterone (P) levels increased on Months 3 and 11, and 17 alpha-OH-progesterone (17 alpha-OHP) levels peaked on Month 3, whereas 20 alpha-OH-progesterone (20 alpha-OHP) levels increased sharply after Month 6. PGFM indicated peaks on Months 2 and 11. F and PMSG levels peaked on Months 2 and 3. From factor analysis, Month, E1, E2 and 20 alpha-OHP were discriminated as Factor 1, increasing with the progress of pregnancy, PMSG, 17 alpha-OHP and P as Factor 2, showing a relation with the secondary corpus luteum, and PGFM and F as Factor 3, affecting PGFM change on Month 2 by F. P also related to both Factors 1 and 3, showing an inverse relation against PGFM. In conclusion E1, E2 and 20 alpha-OHP contained in Factor 1 were suggested to be important especially as parameters of placental function after Month 6.     

Human chorionic gonadotropin influences ovarian function and concentrations of progesterone in prepubertal Angus heifers

The effects of administering hCG on subsequent ovarian structure dynamics and concentrations of progesterone in prepubertal heifers were evaluated. Forty-seven purebred Angus heifers were assigned randomly to 1 of 3 treatments: 1) 100 μg of injection of GnRH (GnRH; n = 16); 2) 1,000 IU injection of hCG (H1000; n = 16); and 3) 500-IU injection of hCG (H500; n = 15). From d -1 to 9 relative to treatment (d 0), daily blood samples were collected to determine concentrations of progesterone and ovaries of each heifer were examined daily by transrectal ultrasonography. Diameter of all follicles ≥ 4 mm and all luteal structures were mapped. Disappearance of the largest follicle occurred within 2 d in a greater percentage (P < 0.05) of all heifers in the H1000 treatment (87.5%) compared with GnRH heifers (43.8%), whereas H500 heifers (73.7%) were intermediate. A new luteal structure formed after follicle disappearance in a greater (P < 0.05) percentage of all heifers treated with H1000 (87.5%) and H500 (73.7%) heifers compared with that in GnRH-treated heifers (18.8%). The largest follicle present in ovaries of H1000 and H500 heifers was smaller (P < 0.05) than that of GnRH heifers from d 2 to 5. Heifers treated with H1000 (1.72 ± 0.19 ng/mL) had peak concentrations of progesterone on d 6 that were greater (P < 0.05) than H500 heifers (1.34 ± 0.20 ng/mL), which were greater than heifers treated with GnRH (0.31 ± 0.19 ng/mL). The mean volume of luteal tissue was greater (P < 0.05) in H1000 heifers (1.54 ± 0.15 cm(3)) than in H500 heifers (1.15 ± 0.15 cm(3)), which was greater (P < 0.05) than in heifers treated with GnRH (0.23 ± 0.15 cm(3)). We conclude that hCG was more effective than GnRH in its ability to cause disappearance of the largest follicle, increase volume of luteal tissue in the subsequent developing luteal structures, and increase concentrations of progesterone in prepubertal heifers. In addition, hCG seems to be more effective when administered at 1,000 IU than at 500 IU.