IU hCG Research Articles

Prolonged glucocorticoid administration affects oocyte morphology in cats (Felis catus) undergoing an ovarian stimulation protocol

While captivity-related stress and the associated rise in baseline glucocorticoid (GC) concentrations have been linked to ovarian quiescence in some felid species, no study has examined the effects of elevated GC on oocyte quality. This study examined the effects of exogenous GC administration on the ovarian response and oocyte quality of domestic cats after an ovarian stimulation protocol. Entire mature female cats were divided into treatment (n = 6) and control (n = 6) groups. Cats in the GC treatment (GCT) group were given 1 mg kg−1 oral prednisolone daily from Day 0–45. All cats (n = 12) were given 0.088 mg kg−1 day−1 progesterone orally from Day 0–37, before treatment with 75 IU eCG im to induce follicular growth on Day 40, followed by 50 IU hCG im 80 h later to induce ovulation. Cats were ovariohysterectomised 30 h after the hCG treatment. Blood samples were collected on Days 0, 10, 30 and 40 (prior to eCG treatment), 80 h after eCG treatment, and on Day 45 for cortisol, glucose, prednisolone, oestradiol, and progesterone analysis. Cortisol concentrations did not differ between treatment groups throughout the study. Mean glucose concentrations were higher in the GCT cats (P = 0.004). Prednisolone was undetectable in all samples. Oestradiol and progesterone concentrations confirmed that the eCG treatment stimulated follicular activity and ovulation in all cats. Following ovariohysterectomy, the ovarian responses were graded (1 = excellent, 4 = poor) and oocytes retrieved from the oviducts. Each oocyte was given a total oocyte score (TOS: using an 9-point scale, 8 = best) based on four parameters: oocyte morphology, size, ooplasm uniformity and granularity, and zona pellucida (ZP) thickness and variation. Ovulation was confirmed in all cats, with a mean of 10.5 ± 1.1 ovulations per cat. Ovarian mass, ovarian response, number of ovulations, and oocyte recovery did not differ between groups. Oocyte diameter did not differ between the groups, but the ZP was thinner in the GCT group (3.1 ± 0.3 μm vs. 4.1 ± 0.3 μm, P = 0.03). The TOS was similar between treatment and control cats, but the ooplasm grade was lower (1.5 ± 0.1 vs. 1.9 ± 0.1, P = 0.01) and there was a tendency for ZP grade to be poorer (0.8 ± 0.1 vs. 1.2 ± 0.2; P = 0.08) in the treatment group. In conclusion, the GC treatment resulted in morphological changes to oocytes collected following ovarian stimulation. Whether these changes would affect fertility warrants further investigation.

Mechanisms of biphasic surges of testosterone in response to human chorionic gonadotrophin (hCG) in stallions

Human chorionic gonadotrophin (hCG) is included in stimulation tests for diagnosis of cryptorchidism and endocrine function of testes in stallions. Although the exact mechanisms are unknown, circulating testosterone (T) concentration shows a clear biphasic increase in response to a single administration of hCG (Tsunoda. et. al. Animal Reproduction Science. 2010; 121S: S165-S167). In the first experiment, concentrations of circulating T and estradiol-17β (E) in response to intravenous administration of 10,000IU hCG were measured. Concentrations of circulating testosterone were also measured in adult Thoroughbred stallions (n=4). In the second experiment, the same experiment was done using adult male golden hamsters (n=45) to figure out mechanisms of biphasic response of T. Adult male golden hamsters were used as a model of stallions, because golden hamsters are typical long day seasonal breeders same as stallions. In the first experiment, blood samples were taken hourly for 6 hours, and daily for 7 days after administration of hCG. Circulating T concentrations in stallions showed a clear biphasic increase within 6 hours and between 2 and 4 days after administration of hCG. Circulating T concentrations at the second peak were significantly higher than those at the first peak. Circulating E concentrations after hCG administration was similar to that of T at the first peak, though the second increase was not detected. Circulating hCG concentrations sharplyincreased just after administration, then followed by an abrupt decline, and low levels were maintained until day 7. It indicated that concentrations of hCG decreased completely when the second peak of T concentrations appeared in circulation. In the second experiment, 25 male golden hamsters were administered 100IU of hCG, and 20 hamsters were administered saline. Blood samples and testes were taken from 5 golden hamsters 5 hours, 24 hours, 3, 5, and 7 days after administration, respectively. Testicular tissues were investigated by real-time PCR for cytochrome P450scc (P450scc), 3β-hydroxy-steroid dehydrogenase (3β-HSD), 17β-hydroxysteroid dehydrogenase (17β-HSD), and receptors of luteinizing hormone (LHR). Testes were also histologically evaluated by hematoxylin-eosin staining. The results of changes in circulating T and hCG concentrations in male golden hamsters were similar to those in stallions. Expression of P450cc, 3β-HSD, 17β-HSD and number of LHR in testes 3 days after administrations of hCG were significantly higher as compared with those at 0 hour. The number of Leydig cells in the histological section 3 days after administration of hCG were also higher compared to 0 hour. The present study strongly suggests that the increase in numbers of LHR and Leydig cells, together with upregulation of steroid enzymes in testes causes the second increase of T plasma concentration after hCG in stallions.

Follicular fluid and plasma concentrations of progesterone following hCG administration before ultrasound-guided ablation of dominant follicles ≥25mm in mares

Ultrasound-guided transvaginal follicle ablation of dominant follicles ≥25mm during the follicular phase results in luteinization and an increase in circulating concentrations of progesterone (Ignácio et al., Theriogenology. 2021; 160:128-133). Although LH receptors are present in early antral follicles of 5mm diameter (Goudet et al., Biol Reprod. 1999; 60:1120-7) and increase after deviation (Bastos et al., Theriogenology. 2014; 82:1131-6), the ablation-induced luteinization rate may be less for smaller-sized dominant follicles <30mm and progesterone may not achieve concentrations >2ng/mL in a timely manner (Mozzaquatro et al., Reprod Domest Anim. 2012; 47:288-92). The objective of the present study was to determine if hCG administered 24h before ablation of dominant follicles <30 or ≥30mm increases the luteinization rate and results in progesterone concentrations >2ng/mL to support pregnancy. Cycling mares were randomly assigned to four ablation groups: 1) ≥25 (ablation of follicles 25-29mm, n=9), 2) hCG+≥25 (2500 IU hCG, i.v. 24h before ablation of follicles 25-29mm, n=11), 3) ≥30 (ablation of follicles ≥30mm, n=13), and 4) hCG+≥30mm (2500 IU hCG, i.v. 24h before ablation of follicles ≥30mm, n=10). Subsequent to PGF2α – induced luteolysis in all mares and administration of hCG to respective groups, D0 marked the day whenthe dominant follicle reached 25-29mm or ≥30mm and follicular fluid was collected. Blood samples were collected daily until D15, fresh embryos were transferred when plasma plasma progesterone (PP4) was >2ng/mL on D7. Follicular fluid progesterone (FFP4) and PP4 were measured by a commercial RIA kit. Wilcoxon test was used to evaluate FFP4. Repeated measures models and Tuckey test were used to evaluate PP4. Regardless of follicle size on D0, mean FFP4 but not PP4 was higher (P<0.05) in mares that received hCG (115.0ng/mL) vs those that did not (53.0ng/mL). Based on PP4 >2ng/mL, luteinization rate was 43 vs 75% in the ≥25 vs hCG+≥25 group (P<0.05) and 77 vs 67% in the ≥30 vs hCG+≥30 group (P>0.05). From D0, daily PP4 gradually increased and on D7 mean concentrations did not differ among the ≥25, hCG+≥25, ≥30, and hCG+≥30 groups, respectively. Regardless of follicle size, embryo transfer resulted in pregnancy rates of 43% (3/7) for the follicle ablation alone and 43% (6/14) for the hCG + ablation groups. In conclusion, administration of hCG before ablation of follicles 25-29mm but not ≥30mm enhanced the luteinization rate with PP4 >2ng/mL at D7 among groups. This was sufficient to support pregnancy after embryo transfer.

HCG versus histrelin acetate: ovulation inducing agents in mares (Equus caballus) in a commercial breeding program in Uruguay

Hormonal control of ovulation timing is an effective tool to optimize reproductive management and reduce costs in equine breeding programs. Few studies have evaluated the efficacy of GnRH analog histrelin as an ovulation inducer agent. This study aimed to compare the efficacy of human chorionic gonadotrophin (hCG) and histrelin as ovulation inducers in young, adult, and old mares during spring and summer. Reproductive records of 321 estrous cycles of 125 mares, mean age 9.8±0.37 years, range 1.5–24 years, between 360-600 kg body weight, and body condition ≥3 (1/5), were evaluated during spring (151 cycles) and summer (170 cycles). Mares were kept in the field and fed alfalfa hay, balanced concentrate feed and ad-libitum water. The ovaries, uterus, and cervix underwent transrectal examination and ultrasonography daily. When only one dominant follicle of 35 mm and endometrial edema compatible with estrus were reached, ovulation was induced. Each cycle was monitored every two to six hours until ovulation, 24h after induction. Cycles were randomly allotted in two groups; G1 (n=106 cycles): 1500 IU hCG (IV); G2 (n=215 cycles): 250 μg histrelin acetate (IM). A total of 119 cycles were from young (<8 years), 93 from adults (8-15 years), and 109 from old mares (>15 years). Cycles of mares ovulating within 24-48hafter treatment were considered to have responded to the inducer, and those ovulating before 24h and after 48h were excluded from the experiment. Inducer agents, season (spring or summer), and mare age were independent factors, and ovulation between 24-48h was considered the dependent factor. Data were analyzed through Fisher's exact test. The significance level was defined as P<0.05. From the 321 induced cycles, 254 (79.1%) ovulated within 24-48h, 20 (6.2%) before 24h, and 47 (14.6%) after 48h. There were no differences (P=0.2414) in the average response time between G1 (37.2h±3,5) and G2 (39.3h±4,5). The ovulatory response between G1 and G2 did not differ in spring (84.1% versus 90.7%, P=0.2627) and summer (79.2% versus 85.4%, P=0.3534). Independent of the inducer, the season did not influence the ovulation response in the different mare ages (spring P=0.1489; summer P=0.2992). No differences (P=0.896) were found between drugs in young and adult mares. However, old mares had a higher percentage of ovulatory response in spring and summer to histrelin (n=76) than to hCG (n=24) (89.5% versus 45.8%, P<0.0001). In conclusion, histrelin is more effective and accurate in inducing ovulation in old mares (>15 years) than hCG throughout the breeding season.

Time to increase in pregnancy-specific protein B following artificial insemination is a direct determinant of subsequent pregnancy loss in lactating dairy cows

Increasing progesterone (P4) during early conceptus development may be crucial for establishment of pregnancy in dairy cattle. The objective of this study was to determine if human chorionic gonadotropin (hCG) at various times after ovulation will increase serum P4 during elongation and increase the chances for, and reduce variability to, initial increase in pregnancy-specific protein B (PSPB) following artificial insemination (AI). Time to PSPB increase was defined as the first day of increase in concentrations of PSPB between d 18 and 28 after ovulation in cows with ≥12.5% increases for 3 consecutive days compared with baseline. Lactating cows (n = 368) synchronized to Double-Ovsynch (first service) or Ovsynch (second or greater service) received one of 4 treatments: no hCG (control), or 3,000 IU of hCG on d 2 (D2), 2 and 5 (D2+5), or 5 (D5) after ovulation. All cows were examined via ultrasound on d 5 and 10 postovulation to determine percentage of cows with hCG-induced accessory CL (aCL) and to quantify and measure all luteal structures. Samples for serum P4 were collected on d 0, 5, 19, and 20 postovulation. The P4 was increased in D2, D2+5, and D5 groups compared with control. The D2+5 and D5 treatments increased aCL and P4 compared with D2 and control. The D2 treatment increased P4 on d 5 after ovulation compared with control. Serum PSPB samples were collected daily from all cows on d 18 through 28 after ovulation for determination of d of PSPB increase. Pregnancy diagnoses were performed via ultrasound examination on d 35, 63, and 100 after ovulation and AI. The D5 treatment reduced percentage of cows with, and increased the time to, PSPB increase. Primiparous cows with ipsilateral aCL had reduced pregnancy loss before d 100 postovulation compared with cows with contralateral aCL. Cows that had PSPB increase >21 d postovulation had 4× greater chances of pregnancy loss compared with cows that had PSPB increase on d 20 or 21. The highest quartile of P4 on d 5, but not on d 19 and 20, was associated with reduced time to PSPB increase. Time to PSPB increase appears to be an important measurement to understand reasons for pregnancy loss in lactating dairy cows. Increasing P4 utilizing hCG after ovulation did not enhance early pregnancy or reduce pregnancy losses in lactating dairy cows.

Impaired luteal progesterone synthesis observed in gilts with PMSG/hCG-induced estrus affects the expression of steroid, prostaglandin, and cytokine receptors in the endometrium and myometrium during the peri-implantation period

Abstract The present study aimed to examine the effect of impaired progesterone (P4) synthesis, observed in gilts with gonadotropin-induced estrus, on the uterine expression of receptors important for pregnancy establishment. Twenty prepubertal gilts received 750 IU PMSG and 500 IU hCG 72 h later, while 18 prepubertal gilts in the control group were observed daily for estrus behavior. Gilts were inseminated in their first estrus and slaughtered on days 10, 12, and 15 of pregnancy to collect endometrial and myometrial tissues for mRNA analysis using real-time PCR. As we previously described, gilts with PMSG/hCG-induced estrus showed decreased luteal P4 synthesis on days 10 and 12 of pregnancy. PMSG/hCG treatment did not affect P4 receptor mRNA expression in either uterine tissue. In the endometrium, a greater mRNA transcript abundance of estrogen receptors (ESR1 and ESR2), androgen receptor (AR), prostaglandin (PG) E2 receptors (PTGER2 and PTGER4), PGF2α receptor (PTGFR), interleukin 6 receptor (IL6R), and tumor necrosis factor α receptors (TNFRSF1A and TNFRSF1B) was detected in gilts with natural than with PMSG/hCG-induced estrus (P&lt;0.05). In the myometrium, the mRNA expression of AR, PTGER2, and PTGFR was lower, while PGI2 receptor (PTGIR) transcript abundance was elevated in the gilts treated with PMSG/hCG as compared with the control animals (P&lt;0.05). In summary, a decreased luteal P4 level during the peri-implantation period in gonadotropin-stimulated pigs affects endometrial and myometrial receptor expression, with the endometrium being more sensitive to impaired P4 synthesis. Whether the observed changes alter uterine receptivity to local and systemic factors remains to be elucidated.

Evaluation of Estrogen Alpha Receptor (ER-Α) and Progesterone-A Receptor (PR-A) Expression in the Oviducts and Uterus of Indonesian Local Rabbits after Induced by Several Method of Pseudo-Pregnancy

The objective of this study was to determine the expression of ER-A and PR-A in the oviduct and uterus of pseudo-pregnant rabbits after induced by several induction methods of pseudo-pregnancy. This study used 30 local rabbits, aged 1 - 1.5 years, body weight of 2.8 - 3.0 kg and clinically healthy. Rabbits were divided into 6 groups (n = 5), namely: K1 (negative control, given 1.5 mL physiological NaCl injection), K2 (positive control, given 100 IU eCG injection and mated with male rabbits, followed by 75 IU hCG injection 3 days later), K3 (given 100 IU eCG injection followed by injection of 75 IU hCG 3 days later), K4 (artificial copulation was performed by inserting a 1 cm long cotton bud into the rabbit’s vagina), K5 (ovulation induction by injection of 50 IU hCG) and K6 (ovulation induction by injection of 5 g GnRH). On the 8th day after treatment, rabbits were euthanized and their oviduct and uterus organs were taken for histological preparation and stained using the immunohistochemical staining. The expression of ER-A and PR-A was valued using the intensity score and analyzed using the Kruskal-Wallis test followed the Mann Whitney U test. The expression ER-A in the infundibulum, ampullary and isthmus of K1 was significantly different (p &lt; 0.05) from K2 - K6 rabbits. The ER-A expression in the endometrium of uterine horn of K1 and K3 was significantly different with K2, K4, K5 and K6 rabbits (p &lt; 0.05). Furthermore, PR-A expression in oviducts of K1 and K3 was lower than that in K2, K3, K5 and K6 (p &lt; 0.05) and the expression in the endometrium, uterine glands and myometrium of K2 - K6 was higher than K1 rabbits (p &lt; 0.05). In conclusion, the hormonal induction could be used to induce pseudopregnancy in local rabbits based on the ER-A and PR-A expressions in oviduct and uterus. HIGHLIGHTS Pseudo-pregnant rabbits are needed for research purposes in the field of reproduction Several methods of induction of pseudo-pregnancy in rabbits have been carried out in this study Pseudo-pregnancy induction cause changes in the concentration of reproductive hormones (progesterone and estrogen) These changes can be detected based on the presence of receptors for these two hormones (PRA and ERα) in reproductive tract of four groups of rabbits and the results were then evaluated GRAPHICAL ABSTRACT

Effect of progesterone infused controlled internal drug releasing (CIDR) device and timing of gonadotropin stimulation using P.G. 600 on reproductive success in ewes bred out of season.

Objectives of this study were to determine effects of exogenous progesterone (via controlled internal drug releasing devices; CIDR) in combination with exogenous gonadotropins (PMSG/hCG) use either at CIDR removal or 1 d before CIDR removal to induce estrus and cyclicity and subsequently enhance the proportion of ewes lambing, lambing rate, prolificacy, and days to lambing in ewes bred out of season. Multiparous ewes (n = 414) were randomly assigned to one of four treatments: untreated (U, n = 122), 7 d CIDR (C, n = 97), 7 d CIDR plus P.G. 600 (240 IU pregnant mare serum gonadotropin [PMSG] and 120 IU human chorionic gonadotropin [hCG]) at CIDR removal (CPG0, n = 97), and 7 d CIDR plus P.G. 600 (240 IU PMSG and 120 IU hCG) 1 d before CIDR removal (CPG-1, n = 98). Rams (n = 15) were joined with ewes immediately after CIDR removal and remained with ewes for a 21 d breeding period. Lambing data were summarized for the first 10 days of the lambing season and overall. Categorical data were analyzed using the GLIMMIX procedure of SAS whereas non-categorical data were analyzed using the mixed procedure. Proportion of ewes lambing in the first 10 d was greater (P < 0.05) for CPG0 and CPG-1 ewes compared with C ewes, which was greater (P < 0.0001) compared with U ewes. Overall proportion of ewes lambing was greater (P ≤ 0.0001) in all treatments utilizing CIDR compared with U ewes, but no differences (P ≥ 0.84) due to P.G. 600 were detected compared with C. Lambing rate in the first 10 d was greater (P < 0.05) for CPG-1 than C, with CPGO being intermediate, and all CIDR-treated ewes being greater than U (P < 0.0001). Overall lambing rate increased (P ≤ 0.0001) in all treatments utilizing CIDR compared with U ewes, but no differences (P ≥ 0.76) due to P.G. 600 were detected compared with C. Prolificacy was similar among all treatments both for the first 10 d of the lambing season (P = 0.86) and overall (P = 0.80). Day of lambing in the lambing season was reduced (P ≤ 0.03) for CPG0 and CPG-1 compared with CIDR-treated ewes, which was reduced (P < 0.0001) compared with U ewes (days 10.6, 9.0, 13.4, and 24.4 ± 0.9 for CPG0, CPG-1, CON, and U, respectively). Though timing of P.G. 600 did not influence results, the combination of CIDR and P.G. 600 enhanced the proportion of lambs born earlier in the lambing season, and incorporating a CIDR with or without P.G. 600 enhanced the overall proportion of ewes lambing and lambing rate in out-of-season breeding scenarios.

Effect of intramuscular injection of human chorionic gonadotropin on endometrium preparation in frozen-thawed embryo transfer cycle: A randomized clinical trial.

Assisted reproductive therapy (ART) has been developed remarkably in these decades; however, the rate of unsuccessful embryo implantation especially in the frozen-thawed embryo transfer (FET) cycles remains high and is reported up to 70%. The current study was designed to compare the effect of intramuscular injection of hCG on endometrium preparation and embryo implantation, in women undergoing FET compared to the control group. This clinical trial was done on 140 infertile women that underwent FET. The study sample was randomly allocated to the intervention group (two 5000 unit ampoules of hCG were injected intramuscularly before the first dose of progesterone administration) and the control group (without hCG injection). In both groups, 4 days after progesterone administration, the cleavage stage embryos were transferred. The study outcomes were biochemical pregnancy, clinical pregnancy and abortion rate. The average age of intervention and control group was 32.65±6.05 and 33.11±5.36 years, respectively. The basic information between two study groups did not differ significantly. The chemical (30% vs. 17.1%, P=0.073, relative risk (RR)=0.57) and clinical (28.6% vs. 14.3%, P=0.039, RR=0.50) pregnancy rates were higher in the intervention group compared to the control group; these higher ratios were only significant in clinical pregnancy rate. Abortion rate was not significantly (P=0.620) different between the intervention and control groups (4.3% vs. 1.4%, respectively). This study showed that intramuscular injection of 10000 IU hCG before the endometrial secretory transformation phase in cleavage-stage embryo, improves IVF cycle outcomes.

Simultaneously administration of cabergoline and PMSG reduces the duration of estrus induction in anestrous bitches.

This study designed a protocol that would combine pregnant mare serum gonadotrophin (PMSG) and cabergoline (CAB) to induce estrus in bitches. Twenty clinically healthy adult and anestrous female dogs were randomly assigned into four groups. The first group was treated with 5.00 μg kg-1 CAB until the onset of proestrus or for 25 days. The second group was treated with 20.00 IU kg-1 PMSG for 5 days and 500 IU human chorionic gonadotropin (hCG) on the 5th day. The third group was treated with 5.00 μg kg-1 CAB for 10 days in combination with 20.00 IU kg-1 PMSG for 5 days and 500 IU hCG on the 10th day. The control group received 1.00 mL of normal saline. Ovarian changes were evaluated ultrasonographically, and the estrus cycle phase was examined by vaginal cytology. Respectively, three, three and four bitches showed clinical signs of proestrus in each treatment group. The intervals between treatment and proestrus for each group were 30.00 ± 3.05, 7.67 ± 1.20 and 13.00 ± 1.20 days, respectively. Two weeks after estrus, the progesterone mean was 14.51 ± 6.24, 19.96 ± 17.16 and 19.12 ± 9.26 ng mL-1 for each group, respectively. In ultrasonography examination, the largest follicle was identified at 15.66 ± 1.33, 11.66 ± 2.40 and 8.75 ± 2.17 days after the onset of proestrus and the largest follicle's size was measured 6.50 ± 0.55, 4.83 ± 1.64 and 7.07 ± 1.49 mm for each group, respectively. Although the combined use of CAB and PMSG reduced the duration of treatment, alteration of the duration or PMSG dosage can be helpful to improve the results.

Low-dose human chorionic gonadotropin supplementation initiated at the onset of ovarian stimulation can improve oocyte quality without impairing endometrial receptivity: Case series

Whether continuous low-dose human chorionic gonadotropin (hCG) supplementation during controlled ovarian hyperstimulation (COH) can improve oocyte and embryo quality is still controversial in clinical practice. We report the first case series of inadvertent COH in luteal-phase stimulation in the presence of endogenous or exogenous low-dose hCG. Patients were diagnosed with infertility. The first two cases had inadvertent COH during preexisting pregnancy, and one of which produced more high-quality embryos (5 vs 1) in the presence of low hCG. Both cases had a live birth. The third case had 7 repeated failures of IVF, during which a total of 55 oocytes were obtained, but only 3 developed into transferable embryo. However, supplementation of 330 IU hCG per day from the onset of COH resulted in the recovery of one high-quality embryo and subsequent delivery of a healthy baby following fresh embryo transfer in eighth attemption. In conclude, supplementation with low-dose hCG from the onset of ovarian stimulation can improve oocyte quality without impairing endometrial receptivity.

Ovulation induction in Caribbean jennies in the tropics

The population of some endangered donkey breeds in tropical regions has declined recently. Therefore, there is an increasing interest to improve the reproductive management of these animals. Ovulation induction agents are often used in equine practice to optimize reproduction techniques. However, limited information exists to support its use in donkeys. Therefore, we compared 2 ovulation-inducing agents (commonly used in equine practice) to hasten ovulation in Caribbean jennies in the tropics. Five cycles of 8 Caribbean jennies were used. Jennies were monitored via transrectal ultrasonography and in 4 estrous cycles they had ovulation induced with either 2,500 IU of hCG or 1.8 mg of deslorelin acetate when a follicle measuring between 27 - 30 mm or &gt; 30 mm in diameter was diagnosed. In the Control-assigned cycle, jennies were not treated with hormones and had spontaneous ovulation. Follicle diameters affected (p &lt; 0.05) the interval to ovulation. When given with a follicle &gt; 30 mm, either treatment reduced (p &lt; 0.05) the interval to ovulation. A higher percentage of jennies ovulated within 48 hours after hCG or deslorelin acetate with &gt; 30 mm follicle (p &lt; 0.05), and with deslorelin acetate with 27 - 30 mm follicle (p &lt; 0.05) than in the Control-assigned cycle. In conclusion, Caribbean jennies in tropics with &gt; 30 mm follicle responded to ovulation induction with hCG or deslorelin acetate, with ovulation occurring within 48 hours.

Single dose of 300 IU hCG in the early luteal phase in superovulated ewes: Effects on corpora lutea, progesterone profile, and embryo recovery

This study aimed to evaluate the effects of hCG treatment during the early luteal phase on ovarian function, progesterone profile, and embryo yield in superovulated ewes. Superovulated sheep were randomly assigned to receive 300 IU hCG i.m. (GhCG, n = 24) or not (GControl, n = 25) at 96 h after the removal of the progesterone (P4) device (D13). Non-surgical embryo recovery (NSER) was performed eight days after P4 withdrawal. Ultrasound evaluations were performed on D13, D14, D16, and D17. Blood samples were collected on D14, D16, and D17. Superovulation scores were recorded based on the number of corpora lutea (CL) as follows: 1 (≤ 2), 2 (3−5), 3 (6−8), and 4 (≥ 9). NSER efficiency, superovulation response, and luteal tissue area were similar in both groups (P > 0.05). Structural luteolysis tended to be higher in GControl (P = 0.07; 47.0 %) while functional luteolysis was similar in both groups (P > 0.05; 0.0 % and 5.9 %). The recovery rate was greater (P < 0.05) in GhCG (89.8 %) compared with GControl (71.0 %), with similar overall ova/embryo numbers observed for both groups (P > 0.05). GhCG showed a higher concentration of animals with a superovulatory response score of 4 (54.5 %; P < 0.05) compared with the lowest scores. Plasma progesterone on D16 was higher (P < 0.05) in GhCG ewes (11.1 ± 1.5 vs 6.9 ± 1.5 ng/mL). In conclusion, the hCG treatment improved circulating P4 and embryo recovery rate, tended to maintain luteal functionality, and thus constitutes an additional tool for improving embryo yield in superovulated ewes.

Effects of intracerebroventricular and intravenous administration of Kisspeptin-54 and Gonadotropin-releasing hormone agonist in rats with ovarian hyperstimulation

PurposeWe aim to determine the effect of local and systemic administration of kisspeptin-54 on ovarian hyperstimulation. MethodsImmature female rats were used. In order to generate the ovarian hyperstimulation model, 50 IU PMSG was administered for 4 consecutive days and a single dose of 25 IU hCG was administered to all groups except for the sham group. To synchronize the sham group, a single dose of 10 IU PMSG followed by 10 IU hCG (48 h later) was applied. Kisspeptin-54 and gonadotropin-releasing hormone (GnRH) agonists were administered 48 h after hCG injection. While intracerebroventricular injection is performed with stereotaxic surgery, Intravenous administration was from the tail vein. Ovarian weights were measured. FSH, LH, estrogen and progesterone hormones were detected in serum by ELISA. VEGFa, IL-1β, TNF-α, MCP-1 immunohistochemical staining was performed on the ovaries and hypothalamus and their optical densities were determined with Image J. Kiss1R mRNA expression was determined by qRT-PCR. ResultsOvarian weights increased significantly in the OHSS group and the systemic GnRH agonist group. The optical densities of VEGFa, IL-1β, TNF- α and MCP-1 immunoreactivity showed us that both local and systemic applied kisspeptin-54 attenuates the level of investigated inflammation parameters in the ovaries. Moreover, local administration of kisspeptin-54 has been shown to enhance the level of Kiss1R mRNA in both the ovaries and the hypothalamus. Conclusion(s)Local and systemic administration of Kisspeptin-54 as a post-treatment reduces inflammation parameters in the ovaries. These findings promote the potential use of kisspeptin-54 on OHSS.

Evaluation of hCG as gonadotropic support to timed embryo transfer protocol in beef cattle

This study compared the reproductive performance of embryo recipients treated with a timed embryo transfer (TET) protocol using human chorionic gonadotropin (hCG) or equine chorionic gonadotropin (eCG). On a random day of the estrous cycle (Day −10) indicus-taurus recipients (n = 341; 194 nulliparous and 147 multiparous cows) with a body condition score between 3.0 and 4.0, were submitted to the TET protocol consisting of an intramuscular (i.m.) injection of 2.0 mg estradiol benzoate (EB) and the insertion of intravaginal progesterone (P4) device that remained until Day −2.5. On the same day (−2.5), the recipients received i.m. 150 mg D-cloprostenol and 1 mg estradiol cypionate and were randomly divided into two groups: the eCG group (n = 179), in which females received i.m. 300 IU eCG and the hCG group (n = 162), in which females received 150 IU hCG. Then, estrus intensity and the diameter of the dominant follicle (DF) were monitored on D0 and the quality of the corpus luteum (CL) (B mode and color Doppler) was assessed on D7 to select recipients eligible for receiving the transfer of an embryo produced in vitro. Pregnancy diagnosis was assessed 23 days after the transfer. Continuous data were analyzed by ANOVA using a mixed-effects model and Tukey's test. The rates were analyzed using a logistic regression model. The diameter of the DF on day 0 of the TET protocol was influenced by the interaction between gonadotropic treatment and category (P = 0.01), and nulliparous recipients treated with hCG had the smallest diameter. Treatment with hCG and eCG resulted in a high rate of estrus expression; however, the proportion of females with a high-intensity of estrus was higher in the hCG group (79.84 vs. 68.61%, respectively; P = 0.03). The utilization rate (recipients with CL) showed a tendency (P = 0.06) to be influenced by the interaction between gonadotropic treatment and category, wherein nulliparous recipients treated with hCG exhibited a lower utilization rate than the other groups. The diameter, perimeter, and area of the CL were similar (P > 0.1) in all groups. However, the hCG group resulted in CL with a better Doppler evaluation score (P = 0.04), central blood flow (P = 0.03), and tendency towards greater peripheral blood flow (P = 0.08). The rates of conception (32.00% hCG vs. 35.10% eCG; P = 0.46) and pregnancy (24.69% hCG vs. 29.61% eCG; P = 0.20) were similar between the hCG and eCG groups. However, an interaction between the gonadotropic treatment and category revealed lower conception (P = 0.01) and pregnancy rates (P = 0.001) in nulliparous recipients treated with hCG. Treatment with hCG resulted in a greater intensity of estrus expression and CL with a higher Doppler score, which determined rates of utilization, conception, and pregnancy similar to conventional protocols using eCG. However, nulliparous recipients treated with hCG exhibited a lower overall reproductive rate.

Effect of Dosage on Combination Ethylen Glycol and Dimethyl Sulfoxide as Cryoprotectant using Vitrification on Viability of Mice (Mus musculus) Blastoscyst Post Warming

Cryopreservation is a method of reproductive technologythat is useful for embryo and preservation creaturesthat are almost extinct. Cryoprotectants are often used for vitrification are PROH, DMSO and EG becausethey have good and efficient results. The study objectivewas to know does the viability of mice blastocystincrease with the combination of intracellularcryoprotectant ethylene glycol (EG) and dimethyl Sulfoxide (DMSO) in vitrification. This study used mice oocytes thencarried out in vitro fertilization and cultureduntil reaching the blastocyst stage. The mice weresuperovulated using 5 IU PMSG and 5 IU hCG after 48hours, then monomatting using infertile males. Egg cellsare taken by tearing the fertilization bag andwashing using MEM. Embryo is cultured so that it reaches the blastocyst phase and is then divided into 4groups: vitrified using 30% EG, 30% DMSO, 20% combination EG + 10% DMSO and 10% P4 combinationEG and 20% DMSO. The four groups were inserted intothe hemi straw which has been modified with 0.25berurn and then put back into the hemi straw with a size of0.5. Hemi Straw which already containedembryos was inserted into a liquid N2 container with a temperature of -196. Warming was done after theembryo was vitrified for a week. After being removed from the container, heating is done at roomtemperature. The embryo hemi straw was inserted into the sucrose level alternately, i.e. 0.25 M, 0.5 M and1M respectively for 2 minutes. Warming process to removecryoprotectants that are in the embryo.Examination of the viability of blastocyst was done usingan inverted microscope. The results of this studythat there were significant differences between groupsusing combination cryoprotectant EG 20% and DMSO10% (p&lt;0.05) with 84% viability. The combination of EGand DMSO could increase the viability of miceblastocyst. The ethylene glycol 20% and dimethyl Sulfoxide 10% was the best concentration combinationbased on this research.

Sperm quality of artificially matured shortfinned eel is not affected by human chorionic gonadotropin dose and route of administration.

BackgroundAcquisition of high quality sperm is key to the artificial propagation of eels in captivity, but fertility drugs are expensive and repeated handling is stressful to the fish. An interrupted treatment regime (an initial hormone injection to stimulate spermatogenesis, followed several weeks later by weekly booster injections to induce sperm maturation) for acquisition of sperm in captive male eels has promise for high sperm quality on the one hand, and animal welfare benefits on the other. To further develop this approach for shortfinned eel, Anguilla australis, we evaluated the efficacy of (i) different initial doses of human chorionic gonadotropin (hCG) and (ii) route of administration.MethodsMale eels were artificially induced to mature with a single injection of 0, 250, 500 or 1,000 IU/fish of hCG, administered either intramuscularly (IM) or intraperitoneally (IP). Sperm maturation was induced with 150 IU hCG/fish from week 5 onwards and sperm collected for evaluation of quality by computer-assisted sperm analysis.ResultsControl males did not mature and hence, sperm could not be retrieved and analysed, but all other treatments were effective in inducing testicular maturation. Milt volume tended to be higher for fish injected IM compared to those injected IP, whereas hCG dose had no effect. Conversely, the concentration of spermatozoa tended to be higher for several sperm collection time points in IP-injected than in IM-injected fish. Sperm quality, represented by percent motility, percent progressive motility and curvilinear velocity, was equal in fish given an initial dose of 250 IU hCG to those given higher initial doses of hCG.ConclusionsWe recommend that an initial dose of 250 IU hCG/fish be administered to induce spermatogenesis in male A. australis, and, after a period of 4–5 weeks, weekly booster injections of ∼150 IU hCG/fish be administered in the day prior to sperm collection; both routes of administration (IM or IP) are equally effective. We contend that an interrupted treatment regime has notable benefits for induced maturation in male anguillids, as it reduces fish handling and manipulation and reduces the resources required to produce high quality sperm.

Granulocyte colony stimulating factor (GCSF) did not affect the fertility outcomes in women with unexplained infertility after intrauterine insemination: a randomised clinical trial

This study aimed to evaluate the effect of granulocyte colony stimulating factor (GCSF) on fertility outcomes in women with unexplained infertility after intrauterine insemination (IUI). This study is a randomised clinical trial that was conducted on unexplained infertile women referred to infertility Clinic in Yasuj, Iran. All participants were stimulated by letrozole, clomiphene and HMG during the cycle. When at least one follicle was greater than 18 mm, 5000 IU hCG intramuscularly was administered for ovulation induction and then IUI was performed 34–36 hours later. In GCSF group, 300 μg GCSF subcutaneously administrated in two days after IUI. Control group received routine IUI. The main outcome measures were biochemical pregnancy, clinical pregnancy, abortion and ongoing pregnancy rate. Our results showed no statistically significant difference in the biochemical pregnancy (16.3% vs. 12.2%; P=.56), clinical pregnancy (16.3% vs. 8.2%; P=.21), abortion (0 vs. 2.04%; P=.55) and ongoing pregnancy rates (8.2% vs. 14.2%; P=.32) between the control and the G-CSF groups. The results of the current study suggest that systemic administration of 300 μg GCSF in the two days after receiving IUI in patients with unexplained infertility does not offer any beneficial clinical related implantation, and pregnancy rates. IRCT registration number: IRCT20160524028038N4 Impact statement What is already known on this subject? Unexplained infertility means to the inability of couples in pregnancy after a year without obvious male and female infertility factors. Some patients have not been justified in endometrial function, which leads to the defect of the dialogue between the foetus and endometrium and may lead to implantation failure. Granulocyte colony stimulating factor is produced at the maternal-foetal interface during embryo implantation and is the main part of the uterine-cytokine network that is needed to create and maintain pregnancy. What do the results of this study add? The current study suggests that systemic administration of 300 μg granulocyte colony stimulating factor in the two days after receiving IUI in patients with unexplained infertility does not offer any beneficial clinical related implantation and pregnancy rates. What are the implications of these findings for clinical practice and/or further research? These results may not be generalised to all patients and more randomised controlled trials are needed for the comparison of granulocyte colony stimulating factor effects on women with thin and normal endometrial thickness, efficacy, side effects, and pregnancy outcomes of the intrauterine perfusion versus systemic subcutaneous administration of granulocyte colony stimulating factor in unexplained infertility treated with intrauterine insemination.

Hormonal profile in early luteal phase after triggering ovulation with gonadotropin-releasing hormone agonist in high-responder patients.

The major limitations associated with gonadotropin-releasing hormone agonist (GnRHa) triggering are inferior clinical outcomes in fresh embryo transfer cycles caused by luteal phase insufficiency following the GnRHa triggering. We included 153 high-risk patients in this study. In group I, the patients received gonadotropin-releasing hormone agonist (GnRHa) trigger + 1,500 IU human chorionic gonadotropin (hCG) support on the oocyte pick-up (OPU) day; in group II, the patients had a dual trigger (GnRHa + 1,500 IU hCG); and in group III (control), 10,000 IU hCG trigger was prescribed for the final oocyte maturation. The levels of LH, estradiol, and progesterone were evaluated in serum on the stimulation starting day, day 6 of stimulation, on the day of the trigger administration, OPU day, days 3 and 5 post-OPU, and day 14 post-ET, as well as in follicular fluid. Progesterone concentration was significantly lower in group I on OPU+5 compared to the hCG group (I vs. III, р = 0.0065). Progesterone levels were significantly lower in group II in serum on OPU+5 compared to groups I and III (I vs. II, р = 0.0068; II vs. III, р = 1.76 × 108). The progesterone levels were significantly higher in follicular fluid in group III compared to the study groups (I vs. III, р = 0.002; II vs. III, p = 0.009). However, no significant differences in clinical outcomes were found between the groups. Then, we divided all women into pregnant and non-pregnant groups and found that estradiol (p = 0.00009) and progesterone (p = 0.000036) on the day of the pregnancy test were significantly higher in the pregnant women group. Also, progesterone on OPU day was significantly higher in the non-pregnant group (p = 0.033). Two cases of moderate ovarian hyperstimulation syndrome (OHSS) late-onset occurred in group I (3.5%, 2/56), no case of moderate/severe OHSS late-onset in group II, and three cases of moderate late-onset in group III (5.7%, 3/53). The low-dose hCG supplementation improves the luteal phase insufficiency after GnRHa triggering, which is confirmed by the comparable pregnancy rates in fresh transfer cycles between the groups. However, low-dose hCG carries a similar risk of OHSS as the full dose of hCG in high-responder patients.

Superovulation of high-producing Holstein lactating dairy cows with human recombinant FSH and hMG

Superovulation of high-producing dairy cows is a challenging subject in dairy farms with respect to the cost, dose and type of gonadotropin. The objectives of this study were to compare three gonadotropin products: Folltropin-V® (highly purified FSH with porcine origin), Cinnal-f® (recombinant human FSH) and Menotropins® (hMG) for superovulation in high-producing Holstein lactating dairy cows and to investigate the pregnancy outcomes achieved following transferring embryos recovered from donors treated with different gonadotropins. Healthy high-producing Holstein lactating dairy cows (n = 30; milk production: 46.35 ± 8.78 kg; parity: 2–4; days in milk: 80–130 days) without any puerperal problems were selected as donors. On Day 10 after estrus (Day 0 of superovulation), donors (10 cows in each experimental groups) received Folltropin-V® (400 mg NIH, dissolved in 20 ml), Cinnal-f® (20 vials; each vial of 1 ml contains 75 IU Follitropin alfa) and Menotropins ® (20 ampules; each ampule of 1 ml contains 75 IU FSH and 75 IU LH), administered twice daily, in decreasing doses (4,4; 3,3; 2,2; 1,1 ml), over 4 days. On Day 2 of superovulation, donors received 3 doses of prostaglandin F2α analogue, 6 h apart. They were inseminated twice with a frozen semen at 12 and 24 h after standing estrus. Concurrent with the second insemination, donors received 2500 IU hCG (Karma Pharmatech GmbH, Germany). On Day 7 after standing estrus, superovulatory responses (number of CLs, total ova/embryos and transferable embryos) were recorded and Code 1 embryos, recovered from each treated donors, were transferred to synchronized heifers. Pregnancy was detected on Day 30 and 60 after AI. Gestation length, the number and weight of live births were recorded. Data were analyzed using Proc GLM, Proc Mixed and Proc Genmod of SAS. The respective number of corpora lutea, total number of ova/embryos and transferable embryos were not different among donors received Cinnal-f (25.5 ± 3.01, 11.2 ± 2.77, 5.1 ± 0.86), Menotropins (24.0 ± 3.21, 9.0 ± 2.04, 6.3 ± 1.74) and Folltropin-V (20.3 ± 3.21, 8.9 ± 1.90, 5.1 ± 1.16; P > 0.05). Pregnancy rates on Day 30 was similar among treatment groups (P > 0.05). However, pregnancy rates on Day 60 and the number of calves born healthy was less in heifers that received embryos from Cinnal-f treated donors (P < 0.05). In conclusion, Cinnal-f and Menotropins could provide similar superovulatory response to Folltropin-V for superovulation of high-producing Holstein lactating dairy cows.