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Hormonal protocols based on progestogens and equine chorionic gonadotrophin (eCG) are efficient for estrus and ovulation synchronization in ewes. Although eCG is indispensable during seasonal anestrus, it may not be necessary during the breeding season. Thus, we tested the hypothesis that GnRH is effective in replacing eCG during the breeding season allowing satisfactory ovulation rate, luteal function and conception rates after timed artificial insemination (TAI). Ewes (n = 134) with a minimum body condition score of 2.5 (0-5 scale) were treated with intravaginal devices (IVD) containing 60 mg of medroxyprogesterone acetate (MPA) for seven days and received 0.26 mg of sodium cloprostenol at the time of IVD removal. In Exp. 1, at IVD removal, ewes (n = 29) were allocated to three groups: eCG (200 IU at IVD removal; n = 10); eCG+GnRH (200 IU eCG at IVD removal and 4 µg of buserelin 36 h later; n = 10); or GnRH (buserelin 36 h after IVD removal; n = 9). Blood samples were collected 2, 6 and 12 days after TAI moment (54 h after IVD removal), for progesterone (P4) analysis. In Exp 2, the ewes were allocated to eCG (n = 10) or GnRH (n = 10) groups, as above described, and ovulation moment was evaluated 54, 66 and 78 h after IVD removal. In Exp 3, TAI was performed in ewes from eCG (n = 45) and GnRH (n = 40) groups using 100 × 10 6 motile spermatozoa from a pool of semen collected from four rams. In Exp. 1, based on P4 levels, we confirmed that all the ewes ovulated (29/29) and there was no significant effect of group (P = 0.89) or group x day (P = 0.18) on P4 concentration, being observed a significant effect of day (P = 0.0001). In Exp. 2, the maximum DF diameter (P = 0.26) and ovulation moment (P = 0.69) did not differ between groups. In Exp. 3, pregnancy rate was significantly lower (P = 0.02) in GnRH (22.5 %; 9/40) compared to eCG (46.7 %; 21/45). The results indicate that, although ovulation and luteal function were not altered after eCG, eCG+GnRH or GnRH treatment, GnRH alone before TAI cannot be used to replace eCG treatment during the breeding season., Competing Interests: Declaration of competing interest The authors declare no conflicts of interest., (Copyright © 2024 Elsevier Inc. All rights reserved.)

This study determined the effects of administering a glycoprotein with equine chorionic gonadotropin (eCG)-like activity (eCG-like) on corpus luteum (CL) area, serum progesterone concentrations, incidence of multiple ovulations (MOV), estrus expression rate (EER), and pregnancy to timed AI (P/TAI) in Angus cattle synchronized with a 5-d Co-Synch protocol. On Day -8, cattle were body condition scored (BCS), and received a 1.0 g progesterone intravaginal device (IVD) and 100 μg GnRH. On Day -3, the IVDs were removed and 500 μg cloprostenol was administered intramuscularly (i.m.). Cattle were randomly assigned into one of two groups: eCG-like (heifers, n = 232, primiparous, n = 148, and multiparous cows = 485; 300 IU (heifers) and 400 IU (cows) eCG-like i.m. on Day -3), or Control (heifers, n = 240, primiparous, n = 151, and multiparous cows, n = 478; no eCG-like). On Day -2, cattle received a second dose of 500 μg cloprostenol, and on Day 0, 100 μg GnRH was given concurrently with TAI. Estrus expression rate was assessed by observing the tail paint rubbed off in a subset of heifers (n = 372) and all cows on Day 0. Transrectal ultrasonography was used to evaluate the presence of CL on Day -8 and to diagnose P/TAI on Day 30-35. In a subset of cattle (heifers = 194 and multiparous cows = 87), CL area, serum progesterone concentrations, and incidence of MOV were evaluated on Day 7. Heifers, primiparous, and multiparous cows were analyzed separately. Treatment with eCG-like did not affect (P > 0.1) EER in heifers. Estrus expression rate was increased (P ≤ 0.03) in primiparous (68.9 % vs 45.0 %) and multiparous (75.5 % vs. 68.8 %) cows treated with eCG-like compared with Controls. Pregnancy/TAI was increased (P < 0.01) in heifers (65.2 % vs 48.3 %) and primiparous cows (48.3 % vs. 35.1 %) treated with eCG-like than Controls. In multiparous cows with a BCS ≤4 P/TAI was increased (P = 0.03) in the eCG-like group (47.7 %) than the Control group (34.8 %) but was similar (P > 0.1) between treatment groups in multiparous cows with a BCS ≥4.5. The eCG-like treatment increased (P < 0.05) CL area in heifers and multiparous cows and tended (P = 0.10) to elevate serum progesterone concentrations only in heifers. However, it did not affect (P > 0.1) the incidence of MOV in heifers and multiparous cows. Glycoprotein eCG-like administration increased fertility in heifers and primiparous cows, but in multiparous the effect of eCG-like on fertility was associated with BCS., (Copyright © 2024 Elsevier Inc. All rights reserved.)

The aim of the current study was to evaluate the effect of a novel recombinant eCG (reCG) on pregnancy rates to AI (P/AI) in suckled beef cows of different breeds that were synchronized with an estradiol/progesterone (P4)-based protocol for fixed-time AI (TAI). In experiment 1, 1244 Bos taurus suckled cows were used. On Day 0 all cows received an intravaginal P4 device (600 mg P4) and 2 mg of estradiol benzoate. On Day 7, devices were removed, and all cows received 0.150 mg of D-cloprostenol plus 1 mg of estradiol cypionate and were randomly divided to receive 140 IU or 105 IU of reCG or no reCG treatment (controls) at that time. Cows were tail painted for estrus detection and those in estrus by 48 h after P4 device removal were inseminated; whereas those not showing estrus were also inseminated and received GnRH at the same time. In experiment 2, 818 Bos taurus x Bos indicus crossbred suckled cows received the same FTAI protocol used in Experiment 1. Cows were randomly divided at the time of P4 device removal into 4 groups to receive 140 IU, 105 IU or 84 IU of reCG or no reCG treatment. In experiment 3, 345 Bos indicus suckled cows were submitted to the same FTAI protocol as those in previous experiments and were randomly divided into three groups to receive 140 IU or 105 IU of reCG, or 300 IU of serum derived eCG (PMSG). In Experiment 1, estrus rate and P/AI was greater (P < 0.05) in cows treated with reCG (79.9 and 53.5 %, 76.9 and 52.3 % for the 105 UI and 140 UI reCG groups, respectively) than those in the control group (69.9 and 44.4 %, respectively). In Experiment 2, cows treated with reCG tended (P < 0.1) to achieve a greater P/AI than control cows (38.6 %, 37.1 %, 36.2 % and 28.2 % for those receiving 84 IU, 105 IU,140 IU of reCG, and those in the control group); but when P/AI of all cows treated with reCG was contrasted to that of control cows, the difference was significant (P < 0.01). In Experiment 3, P/AI in cows treated with 84 IU of reCG (54 %) did not differ from that of cows treated with serum derived eCG (59 %) but both were greater (P < 0.05) than cows treated with 105 UI of reCG (41 %). In conclusion, treatment with reCG improved fertility in suckled Bos taurus and Bos taurus x Bos indicus beef cows. In suckled Bos indicus cows, although treatment with reCG and serum derived eCG were comparable, the higher dosage of reCG was detrimental to their P/AI., Competing Interests: Declaration of competing interest The authors declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research reported., (Copyright © 2024 Elsevier Inc. All rights reserved.)

This study aimed to evaluate the effects of different doses of equine chorionic gonadotropin (eCG; 200 and 300 IU) administered at the end of a fixed-time artificial insemination (FTAI) treatment protocol on ovulation, pregnancy, and twin rates in Bos taurus beef heifers. In addition, pregnancy losses in heifers with singleton and twin pregnancies were determined. A total of 2382 Angus heifers treated with a 6-day estradiol/progesterone-based protocol for FTAI (J-Synch protocol) were randomly allocated to two experimental groups to receive 200 or 300 IU of eCG administered intramuscularly at the time of intravaginal progesterone device removal; FTAI was performed from 60 to 72 h after device removal. The pregnancy rate did not differ (P = 0.89) between the 200 and 300 IU eCG groups. The number of corpus luteum induced by both eCG doses was determined by ultrasonographic examination 14 days after insemination and those treated with 300 IU of eCG had a greater double ovulation rate (P < 0.05). In addition, 300 IU eCG treated heifers had a higher twinning rate on day 30 of gestation (P < 0.05) and parturition (P < 0.05). Pregnancy losses from 30 days of gestation to calving did not differ between heifers treated with 200 and 300 IU of eCG (P = 0.70). However, regardless of the experimental group, heifers bearing twins had greater pregnancy losses than heifers with singletons (P < 0.05). In conclusion, reducing the dose of eCG from 300 to 200 IU under FTAI treatment protocol decreases double ovulation and twinning rates, maintaining a similar pregnancy rate in heifers. Nulliparous cows carrying two fetuses suffer greater pregnancy losses than cows with singletons., Competing Interests: Declaration of competing interest The authors declare no conflicts of interest that could be perceived as influencing the impartiality of this study., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Background: Equine chorionic gonadotropin (eCG) induces super-ovulation in laboratory animals. Notwithstanding its extensive usage, limited information is available regarding the differences between the in vivo effects of natural eCG (N-eCG) and recombinant eCG (R-eCG). This study aimed to investigate the gene expression profiles of mouse ovaries upon stimulation with N-eCG and R-eCG produced from CHO-suspension (CHO-S) cells. R-eCG gene was constructed and transfected into CHO-S cells and quantified. Subsequently, we determined the metabolic clearance rate (MCR) of N-eCG and R-eCG up to 24 h after intravenous administration through the mice tail vein and identified differentially expressed genes in both ovarian tissues, via quantitative real-time PCR (qRT-PCR) and immunohistochemistry (IHC)., Results: R-eCG was markedly expressed initially after transfection and maintained until recovery on day 9. Glycan chains were substantially modified in R-eCG protein produced from CHO-S cells and eliminated through PNGase F treatment. The MCR was higher for R-eCG than for N-eCG, and no significant difference was observed after 60 min. Notwithstanding their low concentrations, R-eCG and N-eCG were detected in the blood at 24 h post-injection. Microarray analysis of ovarian tissue revealed that 20 of 12,816 genes assessed therein were significantly up-regulated and 43 genes were down-regulated by > 2-fold in the group that received R-eCG (63 [0.49%] differentially regulated genes in total). The microarray results were concurrent with and hence validated by those of RT-PCR, qRT-PCR, and IHC analyses., Conclusions: The present results indicate that R-eCG can be adequately produced through a cell-based expression system through post-translational modification of eCG and can induce ovulation in vivo. These results provide novel insights into the molecular mechanisms underlying the up- or down-regulation of specific ovarian genes and the production of R-eCG with enhanced biological activity in vivo.

The present study was conducted to determine if increases in IGF-1 concentration, associated with treatment of ewes with melatonin, has beneficial effects on pregnancy rates when there is induction of estrus in anestrous ewes. A total of 120 multiparous lactating ewes were assigned to three groups (n = 10/group). Ewes of Group 1 were treated with a melatonin implant for 42 days followed by insertion of a controlled internal drug release (CIDR) device for 14 days with administration of equine chorionic gonadotropin (eCG) at day of CIDR removal. The ewes of Group 2 were treated with a CIDR and eCG at the same times as ewes of Group 1 and ewes of Group 3 were assigned to an be untreated control group. Melatonin implantation resulted in an increase in IGF-1 concentrations and lesser estradiol (E 2 ) and triiodothyronine (T 3 ) concentrations. Ewes of Groups 1 and 2 had the greatest progesterone (P 4 ) concentration compared ewes of Group 3. The E 2 :P 4 ratio was less in ewes of Group 1 compared Group 3. Melatonin implantation of ewes resulted in a greater pregnancy rate compared to treatment with the CIDR and eCG which, in turn, had a greater rate than ewes of the control group. In conclusion, melatonin implantation modulates the hormonal milieu including P 4 , E 2 , T 3 and IGF-1 in seasonally anestrous ewes. Increased IGF-1concentrations, as a result of melatonin treatment, are associated with a greater percentage pregnancy rate when there is treatment of anestrous ewes to induce onset of estrus., Competing Interests: Declaration of Competing Interest The authors have no conflicts of interest to declare., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Gonadotropin-releasing hormone agonists (GnRHa) are used as an alternative to human chorionic gonadotropin (hCG) to trigger ovulation and decrease the risk of ovarian hyperstimulation syndrome. GnRHa is less potent at inducing ovarian vascular endothelial growth factor (VEGF), but may also affect endometrial angiogenesis and early placental development. In this study, we explore the effect of superovulation on endometrial angiogenesis during critical periods of gestation in a mouse model. We assigned female mice to three groups: natural mating or mating following injection with equine chorionic gonadotropin and trigger with GnRHa or hCG trigger. Females were killed prior to implantation (E3.5), post-implantation (E7.5), and at midgestation (E10.5), and maternal serum, uterus, and ovaries were collected. During peri-implantation, endometrial Vegfr1 and Vegfr2 mRNA were significantly increased in the GnRHa trigger group (P < 0.02) relative to the hCG group. Vegfr1 is highly expressed in the endometrial lining and secretory glands immediately prior to implantation. At E7.5, the ectoplacental cone expression of Vegfa and its receptor, Vegfr2, was significantly higher in the hCG trigger group compared to the GnRHa group (P < 0.05). Soluble VEGFR1 and free VEGFA were much higher in the serum of mice exposed to the hCG trigger compared to GnRHa group. At midgestation, there was significantly more local Vegfa expression in the placenta of mice triggered with hCG. GnRHa and hCG triggers differentially disrupt the endometrial expression of key angiogenic factors during critical periods of mouse gestation. These results may have significant implications for placental development and neonatal outcomes following human in vitro fertilization., (© The Author(s) 2020. Published by Oxford University Press on behalf of Society for the Study of Reproduction. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

We investigated the capacity for pregnant mare serum gonadotrophin (PMSG) and human chorionic gonadotrophin (hCG) to induce synchronous oestrus and ovulation in the tammar wallaby (Notamacropus eugenii) after follicular suppression with Lucrin® Depot, a one-month GnRH agonist. On Day 0 pouch young were removed (RPY) to reactivate a normal oestrous cycle and then two groups were treated with vehicle (Control; n = 5 and Superovulation (SOvn); n = 5) and two groups were treated with 7.5 mg of Lucrin Depot (Lucrin; n = 6; Lucrin+SOvn, n = 6). On Day 20 RPY the SOvn and Lucrin+SOvn Group received 20 IU of PMSG, which was followed on Day 23 RPY with 500 IU of hCG. The Lucrin+SOvn females underwent a more synchronous oestrus with 5 of 6 mating on Day 26 RPY while the SOvn (n = 5/5) and Control (n = 5/5) Groups copulated over two days, between Day 25-27 RPY and Day 27-29 RPY respectively. Mating plugs were not detected in any of the females in the Lucrin Group by Day 31 RPY. Autopsy on Day 31 RPY confirmed all females in each treatment group had undergone a reactivated cycle as evidenced by the presence of a large corpus luteum (CL) in one ovary. At autopsy the females in the Lucrin+SOvn Group had highly stimulated reproductive tracts, and their ovaries contained many follicles >3 mm; 14 ± 2.1 and 15.3 ± 2.1 follicles >3 mm in the CL-bearing ovary and contralateral ovary respectively. Similarly, females in the SOvn Group had 11.4 ± 2.4 and 17.4 ± 1.9 follicles >3 mm in each respective ovary. Uterine flushing and ovarian histology confirmed that females in Lucrin+SOvn and SOvn Groups had not ovulated, but normal oocytes were present in the follicles. By comparison, the Control Group had ovulated with a single embryo being recovered from the uterus of 4 of 5 females. In contrast to all groups, females in the Lucrin Group showed follicular suppression (all follicles <1.5 mm) and an unstimulated reproductive tract. We conclude that a suppression plus stimulation regimen using Lucrin Depot followed by PMSG and hCG has the capacity to synchronise oestrus, and that 20 IU of PMSG stimulates the development of antral follicles >3 mm in both ovaries. However, a single 500 IU treatment of hCG on Day 23 RPY was not able to induce ovulation in the tammar wallaby., (Copyright © 2020 Elsevier Inc. All rights reserved.)

The present study supports that 5-day short-term CIDR treatments without administration of eCG are equally effective for inducing oestrus behaviour, preovulatory LH discharge and ovulation in sheep than classical protocols based on 14-day treatments plus eCG at CIDR withdrawal. However, the implementation of a 5-day protocol without eCG for fixed-time artificial insemination would be adapted to a later timing of ovulation (p < .05)., (© 2019 Blackwell Verlag GmbH.)

The efficacy of eight combinations of fluorogestone acetate (FGA, 20 or 40 mg as intravaginal device during 11 days), equine chorionic gonadotropin (eCG, 300 or 500 UI injected 48 hr before FGA removal) and prostaglandin F 2α (cloprostenol, 0 or 50 μg injected 48 hr before FGA removal) aiming at induction and synchronization of oestrus and ovulation was evaluated during the anoestrus season in spring and during the breeding season in autumn in adult Beni Arouss goats. Oestrous behaviour was recorded between 12 and 60 hr after FGA removal. Blood samplings allowing to assess onset of the pre-ovulatory LH surge and increase of progesterone as sign of an active corpus luteum were performed, respectively, between 20 and 60 hr and 3, 5, 8 and 15 days after FGA removal. No season-related differences (spring vs. autumn) were observed for oestrous response (95% vs. 93%), pre-ovulatory LH surge (94% vs. 84%) and luteal response after 3-8 and 11-15 days post-treatment (respectively 92% vs. 66% and 92% vs. 98%). The onset of oestrus (21 [13-53] vs. 32 [12-54] hr) and LH surge (26 [20-60] vs. 38 [22-60] hr) occurred significantly later in autumn. FGA (40 vs. 20 mg) in autumn significantly delayed the onset of oestrus (36 [16-54] vs. 23 [12-47] hr) and LH surge (44 [26-58] vs. 33 [22-60] hr). Significant treatment-related differences were recorded for onset of LH surge (earliest for 20 mg FGA, 300 IU eCG, 50 μg PGF 2α ) and onset of luteal phase (latest for 40 mg FGA, 300 IU eCG, 50 μg PGF 2α ). In conclusion, the hormone combinations tested appeared equally effective in terms of oestrous and ovulation rates. Season has influenced significantly the onset of oestrus and LH surge, and the high dose regimen of FGA delayed the ovarian response in autumn., (© 2019 Blackwell Verlag GmbH.)

This study aimed to evaluate the effects of coasting, cabergoline and clarithromycin in a rat ovarian hyperstimulation syndrome (OHSS) model. The 42 female Wistar rats were divided into seven groups: control, OHSS (was given 10 IU of pregnant mare serum gonadotropin for 4 consecutive days from day 22 and 30 IU hCG on the fifth day to induce OHSS ), coasting (hCG was applied on the 27th day after gonadotropin injections and the rats were decapitated on the 28th day), Cabergoline (100 mg/kg/d) and clarithromycin (100 mg/kg/d) were given (on the 26th day) with a short-term supplementation (on the 26th day) and long-term supplementation (from the 22nd to the 26th day) groups. The rats were decapitated on the 27th day. Cabergoline and clarithromycin significantly lowered VEGF-2 levels. Clarithromycin significantly reduced IL-1b and TNF-a and significantly increased IL-10 levels. Clarithromycin may be an effective drug for the treatment of OHSS. Impact statement What is already known on this subject? Ovarian hyper-stimulation syndrome (OHSS) is a self-limited disease, in which vascular endothelial growth factor (VEGF) plays the most important role and has a large clinical spectrum related with increased capillary permeability and fluid retention. Some treatment methods that block VEGF over-expression are used in treatment of OHSS. Clarithromycin is known to suppress the production of some pro-inflammatory molecules such as VEGF, IL-8, IL-1, IL-6 and TNF-a. In our study, we compared the efficacy of coasting, short- and long-term supplementation of clarithromycin and cabergoline on correcting OHSS parameters in an experimental study. What do the results of this study add? As a result of our study, we found that OHSS parameters improved better in early prophylactic treatment regimens. We have shown that clarithromycin may be a more effective treatment agent than coasting and cabergoline. What are the implications of these findings for clinical practice and/or further research? Although our study is important in that it is the first pilot study to show that clarithromycin is effective in the treatment of OHSS, there is a need for larger clinical trials.

The aim of this study was to evaluate the effect of a co-treatment of follicle-stimulating hormone (FSH) plus equine chorionic gonadotrophin (eCG) on serum insulin and insulin-like growth factor 1 (IGF-1) concentrations, superovulatory response, ovulatory rate, and number and production of embryos in Katahdin breed ewes during the non-breeding season. Twenty Katahdin ewes were synchronized with progestagens (CIDR) and assigned to two superovulation treatments (n = 10): (1): ewes treated with 200 mg ewe -1 of FSH from day 5 to 8 after CIDR insertion at decreasing doses every 12 h (FSH group) and (2) ewes treated as FSH group plus 300 IU of eCG on day 5 after CIDR insertion (FSH + eCG group). Estrous behavior was monitored and direct mating was performed. On days - 7 (CIDR insertion), 0 (CIDR withdrawal), and 7 (embryo recovery), blood samples were collected to determine serum hormone concentrations. Co-treatment with eCG (FSH group) did not affect (P > 0.05) serum hormone levels. Superovulation response, ovulation rate, recovery rate, fertilization, and number of embryos were also similar (P > 0.05) between treatments. Compared with FSH group, FSH + eCG ewes had lower (P < 0.05) number of transferable embryos and higher (P < 0.05) number of oocyte and a tendency to increase the number of degenerated embryos (P = 0.07). Overall results suggest that the administration of eCG is not beneficial either to improve the ovulatory response or the amount of transferable embryos in Katahdin ewes superovulated with a protocol using progesterone and FSH at decreasing doses.

This study aimed to elucidate the effects of repeated pregnant mare serum gonadotropin (PMSG) treatment for oestrous synchronization (ES) on ovarian gene expression and reproductive parameters in Xinong Saanen dairy goats, the dominant breed of dairy goat in China. The experiment was carried out at the Research Station of Northwest A&F University (NWAFU), China (34°16'N, 108°4'E). Forty-one does were randomly assigned to groups receiving ES treatments thrice every fortnight (3-PMSG group; n = 19), or ES treatment only once simultaneously with the third ES treatment in the 3-PMSG group (1-PMSG group; n = 22) during middle of the breeding season from late July (14 hr light) until late September (12 hr light). ES treatment was performed via intravaginal insertion of a controlled internal drug release (CIDR) device impregnated with 300 mg progesterone (P4), followed by 300 IU PMSG injections 48 hr before CIDR withdrawal. Oestrus was monitored using vasectomized bucks. Ovaries of three goats in oestrus from both groups were harvested for morphological examination and RNA sequencing (RNA-Seq). Then, all the oestrous goats in the 1-PMSG (n = 21) and 3-PMSG (n = 11) groups were artificially inseminated twice. The 3-PMSG group showed reduced oestrous rate (57.89%), pregnancy rate (31.58%) and litter size (1.17) compared, respectively, with 95.45%, 68.18% and 1.67 for 1-PMSG group (p < 0.05). However, no differences were found in the ovarian morphology between the 1-PMSG and 3-PMSG groups (p > 0.05). RNA-Seq revealed 114 differentially expressed genes (DEGs) in the ovaries of the 3-PMSG group, among which GCG, FSTL3, TET3 and AQP3 were deemed novel and promising candidate genes for regulating fertility. The present study indicates that the three-time PMSG treatment dysregulated several ovarian genes, thereby reducing reproductive performance., (© 2019 Blackwell Verlag GmbH.)

Equine Chorionic Gonadotropin (eCG) previously known as Pregnant Mare Serum Gonadotropin (PMSG) has been used for decades in regulating reproduction in various domestic animal species. Its use necessitates a good measurement of its bioactivity in commercial preparations. The EUROPEAN PHARMACOPEIA (EP 7.0) recommends 5-6 subcutaneous injections in immature female rats for the in vivo bioassay for eCG as in the case for measurement of FSH bioactivity in the Steelman & Pohley assay (1953). This recommendation is in marked contrast with the classical and long-time used PMSG assay of Cole & Erway (1941) that includes only one subcutaneous injection, 3 days before measurement of ovarian weight. As this difference introduces much confusion in the determination of eCG/PMSG bioactivity in commercial sources, we have performed parallel assays of several PMSG preparations, with both protocols. The single-injection protocol takes into account the long half-life of eCG in bloodstream and provokes an ovarian stimulation at lower concentrations than the multiple-injection protocol. As the single-injection protocol also mimicks the protocol used in cattle, it is preferable to the multiple-injection protocol of the current EP., (Copyright © 2019 Elsevier Inc. All rights reserved.)

In most goat breeds, testosterone serum concentration and semen quality decrease during the nonbreeding season. However, bucks reproductive activity may be stimulated with the administration of equine chorionic gonadotropin (eCG). Therefore, the aim of this study was to determine whether the repeated administration of eCG stimulates the reproductive status of bucks during the nonbreeding season. The study was performed with 19 bucks that were assigned to a group that was treated with eCG (GeCG) and an untreated control group (GCon). The GeCG bucks received an initial dose of 800 IU of eCG (Day 0), followed by four doses of 500 IU administered every 5 days beginning on Day 5. Serum testosterone and anti-eCG antibody concentrations, testicular and seminal traits were determined until Day 60. Testosterone concentration (from Day 3 to 21: p < 0.0001), anti-eCG titre (from Day 12 to 44: p ≤ 0.01), percentage of motile spermatozoa (Day 6: p = 0.006 and 14: p = 0.001) and of spermatozoa with progressive motility (Day 6: p = 0.01 and 14: p = 0.002) and the percentage of spermatozoa with functional membrane (Day 6: p = 0.02 and 22: p = 0.008) were higher in GeCG than in GCon bucks. Also in frozen-thawed samples, the percentage of motile spermatozoa tended to be higher in GeCG than that of GCon bucks (p = 0.07). In conclusion, the administration of eCG during the nonbreeding season stimulated the secretion of testosterone and improved fresh and possibly frozen-thawed semen quality. However, it also resulted in an increase in anti-eCG antibody titre., (© 2018 Blackwell Verlag GmbH.)

Two experiments were performed to evaluate the reproductive performance of zebu beef cows treated with different doses of eCG at the end of a progesterone (P4)/estrogen based protocol for timed artificial insemination (TAI). In Experiment 1, suckling Bos indicus Nelore cows (n = 261) received, on day 0, a progesterone (P4) intravaginal device (PD) and an injection of 1 mg estradiol benzoate (EB). On day 8, the PD was removed, 500 μg of cloprostenol was injected, and cows were assigned to one of the following groups: Control (no treatment), 300 (300 IU of eCG), 600 (600 IU of eCG), and 900 (900 IU of eCG). On day 9, all cows received 1 mg EB and TAI performed 54-56 h after cloprostenol injection. A pregnancy diagnosis was done by ultrasound scanning 40 days after TAI, and the number of fetuses and calves was recorded at pregnancy diagnosis and at birth. More cows treated with eCG displayed estrus within 48 h after removal of the PD (42.3% vs. 11.6%, P < 0.01), and ovulated more than one follicle (42%, 58/138 vs. 1.8%, 1/54; P < 0.01). This effect on ovulation rate was dose dependent (P < 0.05). The pregnancy rate was affected only by cow parity (primiparous, 25.3% vs. multiparous, 48.9%; P < 0.01). Twin pregnancy was higher (P < 0.01) in cows treated with eCG (42%, 58/138) than controls (0%, 0/54). However, few cows (33.3%) were able to keep both fetuses intact until birth. For evaluation of ovarian characteristics by B-mode and Doppler ultrasonography, 43 Nelore cows were submitted In Experiment 2 to the same four groups described in Experiment 1. Although no difference (P > 0.1) was observed for size and blood perfusion in the pre-ovulatory follicles, corpus luteum was larger and with greater blood perfusion (P < 0.05) in eCG-treated cows. In conclusion, eCG increased the number of double/multiple ovulations in a dose-dependent manner, induced larger and more vascularized corpora lutea, but did not affect the fertility of cyclic or anestrous cows. Although eCG results in twin pregnancies, most of cows underwet embryo/fetus loss and birth a single calf., (Copyright © 2018 Elsevier Inc. All rights reserved.)

The objective of this study was to evaluate the effect of equine chorionic gonadotropin (eCG) on the estrous response, ovulatory follicle development, ovulation rate, corpus luteum (CL) diameter, concentrations of progesterone (P4) and pregnancy per AI (P/AI) in CIDR-EB treated lactating anovular Nili-Ravi buffalo during the breeding season. Buffalo (n = 87), 60 to 200 Days postpartum were treated with CIDR at a random stage of the estrous cycle (Day 0) and assigned to two groups; with eCG (+eCG; n = 44), or without eCG (-eCG; n = 43). In both groups, each animal received 2 and 1 mg estradiol benzoate (EB) on Day 0 and then on Day 8, respectively. On Day 6, eCG and PGF 2α were administered. On Day 7, CIDR were removed and FTAI was performed at 48 and 60 h later and pregnancy was diagnosed 35 Days post AI. The results revealed that eCG treatment increased (P <  0.05) the CL diameter on 15, 18, and 21 after the ovulation that resulted from imposing the estrous synchronization protocol than when eCG was not administered. Similarly, treatment with eCG resulted in an increased (P <  0.05) mean concentration of P4 as compared with animals not treated with eCG on Days 18 and 21 after the ovulation that occurred subsequent to imposing the estrous synchronization protocol. The ovulation rate was greater in eCG-treated buffalo as compared with those not treated with eCG (93% compared with 74%, respectively; P < 0.05). The P/AI was greater in buffalo treated with eCG than those not treated with eCG (56% compared with 31%, respectively; P < 0.05). In conclusion, the addition of eCG along with EB and P4 based treatments enhanced the estrous response, follicular and luteal function, ovulation rate, concentrations of P4, and P/AI in lactating anovular Nili-Ravi buffalo., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Objective: The aim of the present study was to investigate effects of resveratrol (RSV) over ovarian hyperstimulation syndrome (OHSS) in rat model., Materials and Methods: 24 female Wistar rats (22 days old) were divided into four groups. Group 1 (control group; n = 6) received 0.1 ml intraperitoneal (IP) saline from days 22-26; group 2 (mild-stimulated group; n = 6) received 10 IU pregnant mare serum gonadotropin (PMSG) on day 24 and 10 IU of hCG 48 h later (day 26); group 3 (OHSS group; n = 6) was given 10 IU of PMSG for 4 consecutive days from day 22 and 30 IU hCG on the fifth day to induce OHSS; group 4 (OHSS + RSV group; n = 6) was treated the same as group 3, but received 60 mg/kg RSV 2 h before PMSG injection for 4 consecutive days and 2 h before the hCG injection on the fifth day., Results: Weight gain was highest in the OHSS group. Ovarian weights were lower in the treatment group than OHSS group. Peritoneal fluid VEGF levels were lower for RSV group compared to group 2 and 3. Total VEGF immunoreactivity was higher in OHSS group than group 1, 2 and 4., Conclusion: These results indicate that RSV is beneficial for prevention of OHSS by reducing the increases in body and ovarian weight and VEGF activity. These effects may be mediated by anti-inflammatory, anti-oxidant and anti-angiogenic capacity of RSV., (Copyright © 2018. Published by Elsevier B.V.)

The combined effect of six consecutive timed artificial inseminations (TAIs) on pregnancy rates, following two different synchronization protocols on buffalo heifers, over a period of seven months typically characterized by low breeding performances, were investigated in this study. A total of 2189 TAIs were performed on 1463 buffalo heifers within a large buffalo farm in the south of Italy. Individual animals were allowed to undergo synchronization protocol (either a slightly modified Ovsynch or Progesterone treatment) and TAI until establishment of pregnancy or else for not more than six consecutive times. Semen of seven proven bulls was used throughout the study, which was carried out from March to September of the same year. Therefore, other than the effect given by consecutive TAIs over time, a monthly and a seasonal effect could also be tested, once the entire period was split into a Low Breeding Season (LBS) from March to June, and a Transition to Breeding Season (TBS) from July to September. From the data recorded in this study and the statistical analysis performed, it can be stated that the two protocols for the synchronization of ovulation were similar in efficiency in determining pregnancies with an overall fertility rate of 89.4% when the comparison was run both on a monthly basis or when months were grouped into two different seasons. In addition, an average of 1.83 AI/pregnancy was reported, slightly higher for the Ovsynch when compared to the Progesterone protocol: 1.91 vs 1.70, respectively. Finally, when considering the number of progressive synchronization treatments implemented over time as covariate, neither Ovsynch nor Progesterone treatment significantly affected pregnancy rates following the first of the six synchronization sessions. However, repeating the synchronization procedure, the progesterone based protocol resulted in significantly higher probability of success in terms of established pregnancies during the second and third re-synchronization sessions., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Synchronization programs using progesterone-releasing intravaginal devices that allow for fixed time artificial insemination are still finding increasing application in bovine reproduction. This practice is useful for rationalizing livestock management because an increased number of cows can be inseminated in one session without the need for estrus detection. Although much of the innovation related to the design and development of intravaginal devices for use in cattle took place in the previous century, progress in understanding the physiology of the bovine estrous cycle resulted in shorter treatment durations, a trend which is still continuing. In this competitive market, with little functional differentiation between the existing devices, the shorter treatment duration prompted for optimization of the progesterone content in the device, as the cost of the drug significantly contributes to the price per unit. For CIDR ® a reduction of the progesterone content of about 30 per cent was realized. Price reduction remained an important target for further device development. Next to reduction of progesterone content, cheaper and easier to process materials like polyethylene vinyl acetate (EVA) copolymers have been explored to replace the commonly used silicone elastomers. The reengineering effort of CIDR ® demonstrated that knowledge of release kinetics and insight into gradual depletion patterns in the device is critical for optimization of drug content without compromising performance (blood levels). More recent publications related to the use of alternative polymers like EVA and polyisoprene (IP) indicated encouraging results regarding further reduction of progesterone content. The use of EVA seems most promising, because it is in principle a low-cost polymer available in many grades and this thermoplastic polymer can be processed easily by means of commonly used techniques like injection molding and extrusion. The use of thermoplastic polymers, however, requires insight into the physical-chemical phenomena related to drug dissolution and re-crystallization taking place in the polymer during processing at high temperatures. These aspects, which may critically affect product stability, are often overlooked and are prompting to cover some of the background in this review. Finally, two different innovative approaches are discussed, one related to programable electronic devices for tailored simultaneous drug release and the other is a flexible drug-loaded helix, which is retained very well in several species without causing the usual inflammatory response., (Copyright © 2017 Elsevier Inc. All rights reserved.)

This study aimed to evaluate the effectiveness of hormonal treatments on ovarian activity and reproductive performance in Barki and Rahmani ewes during non-breeding season. Forty-eight multiparous ewes, 24 Barki and 24 Rahmani ewes were divided into two groups, 12 lactating and 12 dry ewes for each breed. Controlled internal drug release (CIDR) device was inserted in all ewes for 14 days in conjunction with intramuscular 500 IU equine chronic gonadotrophin (eCG) at day of CIDR removal. Data were analysed using PROC MIXED of SAS for repeated measures. Breed, physiological status and days were used as fixed effects and individual ewes as random effects. Barki ewes recorded higher (p < .05) total number of follicles, number of large follicles, serum estradiol concentration and estradiol: progesterone (E 2 :P 4 ) ratio compared to Rahmani ewes. Lactating ewes recorded higher (p < .05) number of small follicles and lower concentration of total antioxidant capacity (TAC) compared to dry ewes. Number and diameter of large follicles recorded the highest (p < .05) values accompanied with disappearance of corpora lutea at day of mating. Serum progesterone concentration recorded lower (p < .05) value at day of mating and the highest (p < .05) value at day 35 after mating. CIDR-eCG protocol induced 100% oestrous behaviour in both breeds, but Rahmani ewes recorded longer (p < .05) oestrous duration compared to Barki. Conception failure was higher (p < .05) in Barki compared to Rahmani ewes. In conclusion, CIDR-eCG protocol was more potent in improving ovarian activity in Barki compared to Rahmani ewes, but this protocol seems to induce hormonal imbalance in Barki ewes that resulted in increasing conception failure compared to Rahmani ewes., (© 2017 Blackwell Verlag GmbH.)

This paper reviews that state of our knowledge concerning follicular wave dynamics, monitoring and manipulation. All camelids have overlapping follicular waves in absence of ovulation which is induced by a seminal plasma factor (βNGF). The interval between follicular waves varies. The size of the ovulatory follicle varies between 11 and 25 mm in camels and between in 6 and 13 mm in South American Camelids. The interval between induction of ovulation and next ovulatory follicle is 15 ± 1 day for all camelids. Follicular activity is best monitored by transrectal ultrasonography. Progesterone therapy for 7-15 days seems to suppress follicular dominance but does not completely inhibit follicular recruitment. Combination of estradiol and progesterone seems to provide better control of follicular activity. Both methods have provided variable results in the synchronization of follicular waves. Combination of induction of ovulation with GnRH and luteolysis at predetermined times shows some promise in synchronization of follicular dominance. These synchronization protocols require further investigation in order to provide practical approaches for fixed-time breeding. Ovarian superstimulation with FSH and eCG alone or in combination is somewhat successful. The best results are obtained when treatment is initiated at the emergence of a new follicular wave after induction of ovulation or following treatment with progesterone for 7-14 days. However, response remains extremely variable particularly in terms of ovulation rate and number of recovered embryos. Sources of this variability need to be studied including the effects of season, nutrition, doses and frequency of administration of gonadotropin., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Purpose: DNA methylation is an epigenetic mechanism that plays critical roles during mammalian oocyte and preimplantation embryo development. It is achieved by adding a methyl group to the fifth carbon atom of cytosine residues within cytosine-phosphate-guanine (CpG) and non-CpG dinucleotide sites using DNA methyltransferase (DNMT) enzymes for de novo and maintenance methylation processes. DNMT1, DNMT3A, and DNMT3B play important roles in establishing methylation of developmentally related genes in oocytes and early embryos. The purpose of this study is to identify the effect of superovulation on the expression and subcellular localizations of these three DNMT enzymes in the mouse oocytes and early embryos., Methods: Three groups composed of control, normal dose [5 IU pregnant mare serum gonadotropin (PMSG) and 5 IU human chorionic gonadotropin (hCG)], and high dose [7.5 IU PMSG and 7.5 IU hCG] were created from 4-5-week-old female BALB/c mice. The relative expression and subcellular localizations of the DNMT proteins in the control and experiment groups have been characterized by using immunofluorescence staining subsequently analyzed in detailed., Results: DNMT1, DNMT3A, and DNMT3B protein expression in the germinal vesicle and metaphase II oocytes and in one-cell and two-cell embryos differed significantly when some of the normal- and high-dose groups were compared with the control counterparts., Conclusion: This study has demonstrated for the first time that superovulation alters expression levels of the DNMT proteins, a finding that indicates that certain developmental defects in superovulated oocytes and early embryos may result from impaired DNA methylation processes.

Two experiments were conducted with the aim of determining the effect of equine chorionic gonadotropin (eCG) administration on day 14 after insemination on ovarian response and pregnancy establishment in postpartum anestrous beef cows. In both experiments, cows were subjected to a progesterone- and estradiol-based treatment for fixed-time artificial insemination (FTAI) and were randomly allocated into 4 groups to receive or not receive eCG (400 IU) at the time of device removal and/or at 14 d after FTAI. In experiment 1, from day 14 to 22, daily ultrasonographic determinations were performed to monitor ovarian dynamics, and blood was collected to determine hormone concentrations in 60 cows. In experiment 2, confirmation of pregnancy was performed at 30 and 60 d after FTAI in 1,060 anestrous cows assigned to the same experimental design. Cows that received eCG on day 14 after FTAI showed increases in corpus luteum area (P < 0.01), follicle diameter (P < 0.05), serum progesterone concentrations (P < 0.01), and estradiol-17β concentrations (P < 0.01), compared with cows that did not receive eCG on day 14. Pregnancy rate on day 30 was greater in those cows that received both eCG treatments (ie, at device removal and 14 d after insemination) than in those that did not receive eCG treatment (P < 0.05). In conclusion, eCG administered on day 14 after FTAI increases serum progesterone concentrations during the critical period of pregnancy in anestrous cows, and this second eCG treatment seems to have a positive effect on achieving pregnancy., (Copyright © 2017 Elsevier Inc. All rights reserved.)

The magnitude of ovulation rate (OR) after hormonal induction in sheep should be considered when prolific genotypes are used. We investigated for the first time the effect of the Vacaria allele and its combined effect with the Booroola prolificacy mutation on OR after hormonal treatment during breeding and anoestrous season. A hundred forty-nine Ile de France crossbred ewes, raised in natural pastures in South Brazil, were used to evaluate the OR after treatment with progestagen (MAP) followed or not by equine chorionic gonadotrophin (eCG) treatment (MAP + eCG). During the breeding season, 96% MAP-treated ewes ovulated in comparison to 97% of MAP + eCG-treated females. The double heterozygous carriers (BNVN) presented the higher OR, followed by the single Vacaria (NNVN) and Booroola (BNNN) heterozygous females and least the wild-type (NNNN) ewes. During anoestrus, 96% eCG-treated ewes ovulated, in contrast to 6% treated with MAP alone. The OR of the gonadotrophin-treated females was higher in BNVN and BNNN than NNVN and NNNN ewes. An additive effect in the OR of the two mutations was observed since OR in double heterozygous ewes was similar to the sum of the effects of the alleles of the single heterozygous carrier ewes.

The objective of this study was to compare the efficiency of hormonal treatments on ovarian activity and reproductive performance in anestrous Anglo-Nubian does during the nonbreeding season (February to May). A total of 48 multiparous does were divided into 2 groups (24 lactating does and 24 dry does). In each group, animals were allocated randomly into 2 equal subgroups (12 does each). In the first subgroup, does received a single 18-mg melatonin implant for 42 d followed by a controlled internal drug release (CIDR) device for 19 d in conjunction with 500 IU of equine chorionic gonadotropin (eCG) i.m. on the day of CIDR device removal. The second subgroup received CIDR combined with eCG in parallel with the first subgroup. Melatonin implantation induced a luteotrophic effect, expressed as an increasing number of corpora lutea, increased serum progesterone concentration, and reduced estradiol concentration. Regardless of treatment, dry does showed greater value of progesterone concentration. With the advancement of day of treatment, number of total follicles, small follicles, and medium follicles tended to increase to the greatest values at the day of CIDR device insertion. Furthermore, at day of mating, the numbers of large follicles reached the greatest value, which was associated with the lowest value of the number of corpora lutea. At day of mating, serum progesterone concentration achieved the lowest value, which increased until d 56 of pregnancy. The estradiol:progesterone ratio showed the opposite trend. The detrimental effect of reproductive seasonality, expressed as cessation of estrus behavior and fertile mating during the nonbreeding season, was successfully alleviated by the CIDR-eCG protocol. Furthermore, melatonin implantation in conjunction with the CIDR-eCG protocol enhanced conception rate and fecundity at d 28 of pregnancy and prolificacy at d 56 of pregnancy compared with does that were not implanted. Interestingly, does that failed to conceive did not come to heat again. In conclusion, the beneficial effect of melatonin implantation in conjunction with the CIDR-eCG protocol on the luteotrophic effect was reflected in the increasing number of corpora lutea, increasing progesterone concentration, and decreasing estradiol concentration. Furthermore, conception rate, prolificacy, and fecundity were improved compared with does that were not implanted during the nonbreeding season., (Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The objective of this study was to evaluate the effect of two sources of commercial porcine pituitary-derived follicle-stimulating hormone (pFSH) and pFSH-porcine Luteinizing Hormone (pLH), including equine chorionic gonadotropin (eCG), in ovulatory and embryonic response in Pelibuey sheep. Twenty-four Pelibuey sheep were used and were assigned randomly to four treatments (n = 6): (T1; 200 mg pFSH-Folltropin ® ); (T2; 200 mg pFSH + 300 UI eCG-Folligon ® ); (T3; 250 UI pFSH/pLH-Pluset ® ) and (T4; 250 UI pFSH/pLH + 300 UI eCG). The interval of hours from withdrawal of the device to the beginning of oestrus (BO) was lower (p < .05) in sheep treated with eCG (T2 = 8.0 ± 1.4 and T4 = 10.0 ± 2.8) than in those without eCG (T1 = 12.6 ± 0.6 and T3 = 20.6 ± 2.4). The ovulatory rate (OR) was higher (p < .05) in T1 = 15.5 ± 2.8 and T2 = 15.6 ± 1.4, compared to T3 = 8.1 ± 3.2 and T4 = 11.8 ± 2.8; a significant difference was not shown between them (T1 vs. T2 and T3 vs. T4) when including eCG. The number of non-fertilized oocytes (NFO) was lower (p ˂ .05) in T1 = 0.8 ± 0.4 and T3 = 1.8 ± 1.8, compared to those that included eCG (T2 = 6.3 ± 2.4 and T4 = 2.1 ± 1.2). The number of transferable embryos (TE) was higher (p < .05) when FSH was applied (T1 = 5.8 ± 1.1), compared with (T2 = 2.6 ± 1.1, T3 = 2.3 ± 1.4 and T4 = 2.8 ± 1.5). The commercial treatments (pFSH or pFSH-pLH) in combination with eCG did not improve OR, NFO and TE. However, the exclusive pFSH (Folltropin) treatment presented a higher OR, lower number of NFO and higher number of TE., (© 2017 Blackwell Verlag GmbH.)

Recently, it has been demonstrated that administration of equine chorionic gonadotropin (eCG) in the postpartum period in dairy cows can enhance follicle growth, reduce the interval from calving to first ovulation and increase plasma estradiol concentrations, and, thus, could enhance reproductive performance in a dairy herd when administered on day 6 postpartum. The aim of this study was to investigate the effects of a single dose of eCG between days 9 and 15 postpartum on parameters of reproductive performance in dairy cows. German Holstein cows (n = 1937; primiparous cows: n = 748; pluriparous cows: n = 1189) in a commercial dairy farm were randomly assigned to three experimental groups. Animals within the group eCG received a single dose of 600 IU eCG intramuscularly (i.m.) between days 9 and 15 postpartum followed by an i.m. administration of 500 μg cloprostenol after 14 days. Those of treatment group PG received cloprostenol only between days 23 and 29 postpartum. Cows of the control group remained untreated. Starting on day 49 postpartum, cows were subjected to a Presynch-Ovsynch protocol and inseminated artificially. The impact of application time (days postpartum) of eCG on the intervals calving to first service and calving to conception was statistically not significant. Outcomes of reproductive performance (i.e. first service conception rate, proportion of pregnant cows until 100 and 150 days in milk [DIM], number of inseminations until 150 DIM, calving to first service interval and calving to conception interval) did not differ significantly between treatment group eCG and group PG compared to control group. Regarding postpartum eCG administration, significant interactions between treatment and parity, season, milk yield, and early puerperal disorders, respectively, could not be shown. In conclusion, an eCG treatment of dairy cows between days 9 and 15 postpartum to increase reproductive performance cannot be recommended under the given circumstances., (© 2017 Blackwell Verlag GmbH.)

The effect and underlying mechanism of Bu-Shen-An-Tai recipe on ovarian apoptosis in mice with controlled ovarian hyperstimulation (COH) implantation dysfunction were studied. The COH implantation dysfunction model in mice was established by intraperitoneal injection of 7.5 IU pregnant mare's serum gonadotrophin (PMSG), followed by 7.5 IU human chorionic gonadotrophin (HCG) 48 h later. Then the female mice were mated with male at a ratio of 2:1 in the same cage at 6:00 p.m. The female mice from normal group were injected intraperitoneally with normal saline and mated at the corresponding time. Day 1 of pregnancy was recorded by examining its vaginal smears at 8:00 a.m. of the next day. Fifty successfully pregnant mice were equally randomly divided into 5 groups: normal control pregnant group (NC), COH implantation dysfunction model group (COH), low dosage of Bu-Shen-An-Tai recipe group (LOW), middle dosage of Bu-Shen-An-Tai recipe group (MID) and high dosage of Bu-Shen-An-Tai recipe group (HIGH). Then from day 1, the mice in different groups were respectively intragastrically given corresponding treatments at 9:00 a.m. for 5 consecutive days. The concentrations of 17β-estradiol (E 2 ) and progesterone (P 4 ) were determined by radioimmunoassay (RIA). The ultrastructural changes of ovarian tissues were observed by transmission electron microscope (TEM). The histopathological changes of ovarian tissues were observed by HE staining. The number of atretic follicles and pregnant corpus luteum were also recorded. TUNEL was applied to measure apoptotic cells of ovarian tissues. Western blotting was used to detect the protein expression of apoptosis- related factors like Bax, Bcl-2 and cleaved-caspase-3 in ovarian tissue of mice. The results showed that ovarian weight, the concentrations of E2 and P4, the number of atretic follicles and pregnant corpus luteum, as well as the apoptosis of granulosa cells were significantly increased in the COH group. The ultrastructures of ovarian tissues in the COH group showed that chromatin in granulosa cells was increased, agglutinated, aggregated or crescent-shaped. The focal cavitation and the typical apoptotic bodies could be seen in granulosa cells in the late stage of apoptosis. After the treatment with different doses of Bu-Shen-An-Tai recipe, the ultrastructural changes of ovarian granulosa cells apoptosis were dramatically improved and even disappeared under TEM. Visible mitochondria and mitochondrial cristae were increased and vacuoles were significantly reduced. The lipid dropltes were shown in a circluar or oval shape. The protein expression levels of Bax and cleaved-caspase-3 were decreased, and the expression of Bcl-2 protein was increased after treatment. It was concluded that Bu-Shen-An-Tai recipe can inhibit the apoptosis of ovarian granulosa cells, probably by up-regulating the protein expression of Bcl-2 and down-regulating Bax and cleaved-caspase-3, which contributes to the formation and maintenance of ovarian corpus luteum. It's helpful to promote the embryonic implantation, to reduce embryo loss and ultimately to improve the success rate of pregnancy.

Substantive evidence has indicated that the sympathetic innervation contributes to the regulation and development of ovarian functions. Norepinephrine (NE) is one of the major neurotransmitters contained in the extrinsic ovarian sympathetic nerves and is thought to be a potent moderator of ovarian functions such as steroidogenesis and granulosa cell proliferation or apoptosis. However, the mechanisms of NE regulation of granulosa cell apoptosis in the rat ovary are rare. Real-time PCR and Western blot results show that NE regulates the expression of miR-21 in primary granulosa cells in a dose-dependent manner. Additionally, we found that miR-21 is involved in NE-mediated rat granulosa cells apoptosis and blocks granulosa cell apoptosis by targeting Smad7 , a transforming growth factor-beta-inducible mediator of apoptosis in granulosa cells. In primary granulosa cells, a combined treatment of NE and TGF-β increased apoptosis and decreased miR-21 expression, but increased SMAD7 protein levels. We also demonstrated that NE regulates miR-21 by coupling to α 1A -adrenergic receptor (α 1A -AR). This is the first demonstration that NE controls the reproductive functions by modulating the expression of miR-21 and promoting TGF-β-induced granulosa cell apoptosis. Such NE-mediated effects could be potentially used for regulating the reproductive processes and for treating reproductive disorders., (© 2017 Society for Endocrinology.)

Following the performance of a superovulation protocol, multiple nodules were observed bilaterally in the uterine horns of 31 of 276 (11.2%) C57BL/6 J female mice aged 8.5 ± 0.6 (mean and standard error of mean) weeks. These lesions prevented embryo collection, and the uterine decidual reaction was suspected. Samples of pathological uteri (n = 20) and the normal genital tracts of donors treated with a similar superovulation protocol (control group, n = 10) were collected. Immunohistochemistry was performed to evaluate pancytokeratin, desmin, vimentin, progesterone receptor (PR), estrogen receptor α (ERα), Ki-67, cyclin D3 and c-Myc expression, as well as quantitative polymerase chain reaction to assess cyclin D3, Hoxa-10 and heparin-binding epidermal growth factor-like growth factor (HB-EGF) mRNA expression. The uterine decidual reaction presented a high degree of structural organization and specifically affected the antimesometrial region of the endometrium. The abnormal decidual cells were large polygonal cells that were frequently polyploid or binucleated and strongly positive for desmin. Immunohistochemistry showed higher Ki-67 proliferation index and higher expression of PR and cyclin D3 in decidual cells in the antimesometrial aspect of the endometrium, compared to nondecidualized endometrial stromal cells in the mesometrial aspect of affected uteri, and compared to endometrial stromal cells in healthy uteri. High expression of cyclin D3 and Hoxa-10 mRNA was also observed in uteri affected by the decidual reaction. These results suggest that PR overexpression in endometrial stromal cells, likely due to high progesterone levels, triggers cyclin D3 and Hoxa-10 overexpression, which may be involved in the pathological mechanisms of the mouse uterine decidual reaction.

The aim of the present study was to determine the effects of repeated superovulation on oocyte quality and embryo developmental potential. Female Swiss albino mice were injected with 5IU pregnant mare's serum gonadotropin followed 48h by 10IU human chorionic gonadotropin. Mice were superovulated up to four times with a gap of 7 days between each superovulation cycle. Ovarian weight increased significantly with an increasing number of superovulation cycles. Although the first stimulation cycle resulted in a threefold increase in the number of oocytes, the number of oocytes decreased gradually after subsequent stimulations. Increased cytoplasmic fragmentation, abnormal mitochondrial distribution, aggregation of Golgi apparatus, spindle damage, increased intracellular oxidative stress and a decrease in expression of octamer-binding transcription factor 4 (Oct4) expression were observed in these oocytes. Further, embryos derived from mice subjected to multiple stimulation cycles exhibited a low blastocyst rate, decreased hatching rate and increased apoptosis in blastocysts. In conclusion, the present study demonstrates that repeated superovulation adversely affects mouse oocyte quality by altering the distribution of cytoplasmic organelles, increasing oxidative stress and decreasing Oct4 expression, resulting in poor developmental potential of the embryos.

It is possible to control follicular development in gilts and sows with the use of hormones including the progestogen altrenogest, GnRH, eCG, hCG, and porcine luteinizing hormone (pLH). These hormones can be used to develop protocols for control of estrus with artificial insemination (AI) timed to estrus detection (timed artificial insemination; TAI) or to control estrus and ovulation with insemination at a fixed time without the requirement of estrus detection (fixed-timed artificial insemination; FTAI). In cyclic gilts, double TAI after protocols based on altrenogest and eCG plus hCG administration can achieve a 70% of farrowing rate. Valuable results can be obtained in weaned sows by the utilization of protocols based on eCG administration at weaning and then GnRH or pLH at estrus onset followed by single or double TAI. In cyclic gilts, single or double FTAI regardless of estrus expression can be applied after protocols based on altrenogest administration followed by eCG and then GnRH, hCG, or pLH some hours later; farrowing rates are similar to control animals inseminated at estrus detection. With sows, a protocol based on eCG administration at weaning and hCG, GnRH, or pLH some hours later followed by single or double FTAI can give fertility rates comparable to control animal inseminated at estrus. Most recently, injection or vaginal deposition of GnRH 96 hours after weaning followed by a single FTAI 24 to 30 hours later is resulting in reproductive performance not different to animals subject to multiple inseminations after natural estrus. It is possible to apply FTAI in lactating sows. The protocols are based on eCG during lactation followed by hCG and FTAI. These protocols will induce ovulation during lactation, but pregnancy rates are reduced. However, in the future, a better knowledge on the effect of hormone administration on follicular dynamics during lactation may allow the development of more effective protocols., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Japanese field vole (Microtus montebelli) is a wild-derived rodent and have unique characteristic. Thus, these species have been expected as model animal. This study was performed to develop novel superovulation procedure for Japanese field vole. First, when 30 IU pregnant mare's serum gonadotropin (PMSG) and 30 IU human chorionic gonadotropin (hCG) were administrated 48 hours apart, females showed higher response to hCG compared with three concentrations of PMSG. Second, to effectively induce ovulation on females after vaginal opening, they were mated with vasectomized male instead of hCG administration. Average number of ovulated oocytes using PMSG mating (13.9 ± 1.9 oocytes) was higher than PMSG-hCG (control; 6.9 ± 2.3 oocytes) or PMSG-hCG mating (6.8 ± 0.8 oocytes). Finally, we attempted superovulation using GnRH agonist (GnRHa). With this treatment, we speculated that GnRHa might induce endogenous luteinizing hormone releasing to cause ovulation. Such superovulation was performed with 30 IU PMSG and different concentration of 20% polyvinylpyrrolidone-GnRHa (15, 30, 45, and 60 μg/kg). As results, average number of ovulated oocytes was highest with 30 μg/kg GnRHa (14.5 ± 4.1 oocytes). The numbers of ovulated oocytes of other concentrations were 5.0 ± 1.4 (15 μg/kg), 12.8 ± 2.7 (45 μg/kg), and 8.8 ± 3.7 oocytes (60 μg/kg). Nuclear status of most collected oocytes was the second meiotic division (range, 94.3%-100%). These superovulation procedures will be useful for development of in vitro culture systems and assisted reproductive technologies for not only Japanese field vole but also other voles., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Adult goats (n = 32) were randomly assigned to one of four treatments (n = 8, each): (i) progesterone (P4 ) + equine chorionic gonadotropin (eCG), treated with 25 mg progesterone intramuscularly (i.m.) + 250 IU eCG 24 h later; (ii) cronolone + eCG, treated with vaginal sponges - 20 mg cronolone × 7 days + 250 IU eCG at pessary removal; (ii) P4 + estradiol (E2 ), treated with 25 mg progesterone i.m. + 1 mg estradiol 24 h later; (iv) cronolone + E2 , treated with vaginal sponges - 20 mg cronolone × 7 days + 1 mg of estradiol i.m. at pessary removal. Goats were tested for estrus throughout the presence of a buck. Seven days prior and after treatment, an ovarian ultrasonographic scanning was performed to determine ovarian function and structures. An ultrasonographic pregnancy diagnosis was performed on day 30 post-service. In all groups, 100% estrus response was observed within 96 h post-treatment. While ovulation occurred in 100% of P4 + eCG and cronolone + eCG treated goats, the other groups only depicted 50% ovulatory activity (P < 0.05). Pregnancy rate was higher (P <0.05) in the P4 + eCG and cronolone + eCG groups (88 and 100%, respectively), compared with 38% in P4 + E2 and cronolone + E2 groups. The best treatments were those in which eCG was applied. The P4 + eCG treatment was a pessary-free, cheaper and effective protocol to induce ovulation in goats during the seasonal anovulatory period., (© 2015 Japanese Society of Animal Science.)

The aim of the study was to evaluate the feasibility of pre-selection of high or low responder does prior to the superovulatory protocols. Twenty Saanen does received 800 IU of equine chorionic gonadotropin (eCG) at the end of long-term progestogen treatment. Fourteen days later, a second progestogen protocol associated with a multiple-dose follicle stimulation hormone (FSH) treatment (5 IU/kg of FSH, in six decreasing doses between days 4 to 6 of the protocol) was administered. Transrectal ultrasound was used to assess the follicular status at the beginning of superovulatory treatments, at the oestrous onset and on the seventh day of the oestrous cycle for counting corpora lutea (CL). A significant lower number of CL was obtained in eCG-treated in comparision with FSH-treated does (p < 0.05). A quartic regression was able to explain the relationship between the number of CL in response to both treatments (r(2) =0.50; p < 0.05). Seventy per cent (14 of 20) of does maintained the same ovulatory response (high or low) after treatments. The Kappa (κ = 0.40; p < 0.05) and Spearman (rs = 0.39; p = 0.08) coefficients were able to show a relationship between treatments. Regarding the follicular status, there is a significant relationship between the number of small follicles (r = 0.71; r(2) =0.47; p < 0.01) and total follicles (r = 0.60; p < 0.01) at eCG and first FSH dose with the number of CL. Moreover, it was found a negative relationship between the presence of large follicles and the number of CL in response to eCG treatment (r = -0.44; p < 0.05), but not from FSH (p > 0.05). In conclusion, the screening test with eCG has the potential to identify Saanen does that will better respond to the superovulatory protocol with FSH. In addition, it highlighted the importance of an ultrasound evaluation prior to the beginning of superovulatory treatments with FSH to characterize the follicular status and identify the potential donors of high ovulatory response in MOET programmes in goats., (© 2016 Blackwell Verlag GmbH.)

The current investigation aimed to establish a fixed-time induction of ovulation/ insemination protocol in camels superovulated by different equine chorionic gonadotropin (eCG) regimens during the transition period in Egypt (mid-October to mid-November). Seventeen pluriparous camels, Camelus dromedarius, were used. All females retained controlled intra-vaginal drug releasers (CIDRs) for 13 consecutive days, and at CIDR withdrawal, the camels were randomly divided into three groups. The control group (n = 5) received 1 ml saline intra-muscularly (i.m.), whereas remaining camels were superovulated by 2500 IU eCG either in a single shot (SS, n = 6) or in serial decreasing doses over 3 days (DD, n = 6). Ovarian dynamics were monitored by transrectal ultrasonography at 2-day intervals, and ovulation was induced by 5000 IU hCG i.m. The changes in reproductive hormones throughout the period of the study were determined. The results showed that mean values of total no. of follicles and size of dominant follicles remained low (P < 0.05) in all groups until day of CIDR removal. Thereafter, total follicle no. increased (P < 0.05) in both superovulated groups compared to the control, where the dominant follicles attained the highest (P < 0.05) diameter 12 days after the eCG treatment. Double-ovulation rate was higher (P < 0.05) in SS (50%) and DD (66.6%) groups compared to that of control (0.0%). However, 33.3% of the SS group developed large anovulatory follicles (ø > 25 mm), which did not respond to induction to ovulation. These results elucidate that eCG administration in serial decreasing doses generates a reliable superovulatory response in camels, and ovulation can be blindly induced 12 days after the gonadotropin treatment. This fixed-time hormonal protocol represents a sufficient alternative to conventional day-to-day ultrasonography and would have profound implication for enhanced fertility in dromedary camels by facilitating infield application of embryo transfer technique.

This study evaluated the effect of different doses of eCG (control, 300 or 400 IU) administered at progesterone (P4) device removal in suckled Bos taurus beef cows undergoing a timed artificial insemination (TAI) protocol. A total of 966 cows received a P4 insert and 2.0 mg intramuscular estradiol benzoate at the onset of the synchronization. After 9 days, P4 insert was removed, and 12.5 mg of dinoprost tromethamine and 1 mg of estradiol cypionate were administered, followed by TAI 48 hours later. Then, the cows received one of three treatments as follows: control (n = 323), 300 (n = 326), or 400 IU of eCG (n = 317). A subset (n = 435) of cows in anestrus had their ovaries evaluated using ultrasound at the time of P4 removal and at TAI. Data were analyzed by orthogonal contrasts (C): C1 (eCG effect) and C2 (eCG dose effect). Estrous occurrence (control = 53.7%, 300 IU = 70.6%, and 400 IU = 77.0%) and pregnancy per artificial insemination (control = 29.7%, 300 IU = 44.8%, and 400 IU = 47.6%) were improved by eCG treatment (C1; P = 0.0004 and P < 0.0001, respectively). Furthermore, the cows receiving eCG presented larger follicles at TAI (control = 13.5 ± 0.3 mm, 300 IU = 14.0 ± 0.2 mm, and 400 IU = 15.1 ± 0.3 mm; P < 0.0001; C1). However, there was no effect of eCG dose on any response variables studied (C2; P > 0.15). In conclusion, the eCG treatment administered at the time of P4 removal increased the occurrence of estrus, the larger follicles at TAI, and pregnancy per artificial insemination of suckled B taurus beef cows. Despite the greater occurrence of estrus in noncyclic cows receiving 400 IU of eCG, both eCG doses (300 and 400 IU) were equally efficient to improve pregnancy to artificial insemination., (Copyright © 2016 Elsevier Inc. All rights reserved.)

The aim was to examine the effect of a single intramuscular (i.m.) injection of equine chorionic gonadotrophin (eCG) on Day 3 after oestrus on corpus luteum (CL) development, circulating progesterone and conceptus development in cross-bred beef heifers. In Experiment 1, heifers received: (1) saline, or a single i.m. injection of eCG on Day 3 at (2) 250IU (3) 500IU (4) 750IU or (5) 1000IU. Administration of eCG resulted in increased luteal tissue area and progesterone and oestradiol concentrations compared with controls. In Experiment 2, heifers received (1) a progesterone-releasing intravaginal device (PRID Delta) from Day 3 to 5 or (2) a PRID Delta from Day 3 to 5 plus a single injection of 750IU eCG on Day 3. In vitro-produced blastocysts (n=10 per recipient) were transferred on Day 7 and heifers were slaughtered on Day 14 to assess conceptus development. Administration of eCG reduced the number of short cycles (6.3% vs 31.3%) and increased mean luteal tissue weight (P=0.02). Insertion of a PRID Delta on Day 3 resulted in an elevation (P<0.05) in serum progesterone until removal on Day 5. Administration of eCG at the time of PRID Delta insertion resulted in higher progesterone levels (P<0.05) from Day 10 onwards. Conceptus dimensions were not affected. In conclusion, a single injection of eCG on Day 3 increased CL size and progesterone concentrations and, when given in conjunction with a progesterone-releasing device, appeared to reduce the number of short cycles, presumably due to its luteotrophic nature. The implications of the elevated oestradiol concentrations for embryo quality require further study.

The objective of this study was to validate the use of exogenous gonadotropin (PG600) treatment for stimulating estrus in noncyclic gilts and to compare lifetime productivity of gilts recorded as having natural (NAT) versus PG600-induced (PG600) first estrus in a commercial setting. Prepubertal Camborough gilts ( = 4,489) were delivered to a gilt development unit (GDU) with the goal of delivering known cyclic breeding-eligible females to the sow farm (SF). A boar exposure area (BEAR) was designed to facilitate stimulation and detection of puberty by providing fence line and direct contact (15 min daily) with mature boars over an intensive 28-d period, starting at approximately d 160 (d 0). At d 14, nonpubertal gilts were mixed in new pen groups. At d 23, noncyclic "opportunity" gilts with no record of vulval development and required to meet breeding targets, were eligible for treatment with PG600 to induce puberty. Overall, 77.6% ( = 3,475) of gilts exhibited standing estrus (NAT = 2,654; PG600 = 821) and were eligible for shipping to the SF at approximately 35 d, and 76.6% of gilts that were administered PG600 exhibited the standing reflex within 13 d of treatment. Ultimately, 72.0% of gilts entering the GDU were delivered to the SF as breeding-eligible females. Considering the gilts delivered, a greater proportion of NAT than PG600 gilts were successfully bred ( < 0.001) and had better farrowing rates to first service, and overall farrowing rates (including gilts that returned to estrus and were rebred) were greater for NAT compared to PG600 gilts ( < 0.001) . Farrowing rates at second and third parity were similar between NAT and PG600 gilts; however, at fourth parity, a greater proportion of NAT gilts farrowed. In comparison, considering only gilts served, there was no difference ( > 0.05) in the proportion of NAT and PG600 gilts farrowing a third litter, but a greater proportion of NAT than PG600 gilts farrowed their fourth litter ( < 0.001). There was no difference between NAT and PG600 gilts for litter size at parity 1 through 4 or total pigs born over 4 parities ( > 0.05). A negative correlation ( < 0.0001) was detected between age at puberty and lifetime growth rate at puberty, and growth rate classification affected age and weight at puberty. However, retention rates and total sow productivity to parity 4 were not affected by growth rate classification at puberty.

Superovulation induced by exogenous gonadotropin treatment (PMSG/hCG) increases the number of available oocytes in humans and animals. However, Superovulatory PMSG/hCG treatment is known to affect maternal environment, and these effects may result from PMSG/hCG treatment-induced oxidative stress. 2-Cys peroxiredoxins (2-Cys Prxs) act as antioxidant enzymes that protect cells from oxidative stress induced by various exogenous stimuli. Therefore, the objective of this study was to test the hypothesis that repeated PMSG/hCG treatment induces 2-Cys Prx expression and overoxidation in the reproductive tracts of female mice. Immunohistochemistry and western blotting analyses further demonstrated that, after PMSG/hCG treatment, the protein expression levels of 2-Cys Prxs increased most significantly in the ovaries, while that of Prx1 was most affected by PMSG/hCG stimulation in all tissues of the female reproductive tract. Repeated PMSG/hCG treatment eventually leads to 2-Cys Prxs overoxidation in all reproductive organs of female mice, and the abundance of the 2-Cys Prxs-SO2/3 proteins reported here supports the hypothesis that repeated superovulation induces strong oxidative stress and damage to the female reproductive tract. Our data suggest that excessive oxidative stress caused by repeated PMSG/hCG stimulation increases 2-Cys Prxs expression and overoxidation in the female reproductive organs. Intracellular 2-Cys Prx therefore plays an important role in maintaining the reproductive organ environment of female mice upon exogenous gonadotropin treatment.

The aim of this study was to assess the dose (300 to 600 IU) effects of equine chorionic gonadotropin (eCG) on the preovulatory follicle diameter, growth rate and time of ovulation characterized by echography. The eCG was injected at the end (D0) of the 7-day treatment with a controlled internal device release (CIDR®) and a PGF2α being injected 2 days before the removal of the CIDR® (d-2). The 120 N'Dama female were distributed into five experimental groups. The control group (n = 26) was treated with physiological saline at the removal of the CIDR®, while the animals in the four treated groups received, respectively, 300 IU (n = 25), 400 IU (n = 24), 500 IU (n = 22) and 600 IU (n = 23) of eCG. The diameter of the preovulatory follicle was significantly higher (P < 0.05) in the animals treated with 300 IU (10.1 ± 1.4 mm) than in untreated animals (9.3 ± 1.2 mm). Follicle growth rate was significantly (P < 0.05) higher in treated animals (1.0 ± 0.4 mm/day) than in the control group (0.9 ± 0.4 mm/day). The average interval between the time of eCG injection and ovulation was similar in the non-treated (83.7 ± 14.4 h) and treated animals (79.7 ± 11.9). Treated animals showed a significant increase in the percentage of ovulation (94.7 % compared to 73.1 %) (P < 0.01). Use of eCG contributed towards synchronising the time of ovulation between 72 to 96 h, which would facilitate the use of systematic insemination.

The effects of equine chorionic gonadotropin (eCG) on follicular development and ovulation in cyclic guinea pigs were investigated by histological and immunohistochemical analyses. Three groups of guinea pigs (n=12) were administrated subcutaneously with saline, 20 or 50 IU of eCG, respectively, on cyclic Day 12 (Day 1=vaginal openings). Ovaries were collected at 4 and 8 d after administration (6 animals per group each time). The eCG administration induced significant and distinct morphological changes in the ovaries, as it promoted the luteinization of granulosa cells, but not follicular development. In addition, proliferating cell nuclear antigen (PCNA) and steroidogenic acute regulatory protein (StAR) were immunolocalized specifically in luteinized follicles. Our experiments together indicate that eCG administration can induce follicular luteinization but not superovulation in guinea pigs. The eCG in cyclic guinea pigs functions similar to that of luteinizing hormone (LH), but not follicle-stimulating hormone (FSH).

This study aimed to evaluate the morphological changes in microvascular density and corpus luteum (CL) vascularization in cows treated with eCG during stimulatory and superovulatory protocols. Sixteen cows were synchronized and divided into three groups: control (n = 6), stimulated (n = 4) and superovulated (n =6), one was submitted to estrous synchronization (ES) and received no eCG (control), and those that were submitted to ES and received eCG before or after follicular deviation (superovulation and stimulation of the dominant follicle, respectively). Ovulation was synchronized using a progesterone device-based protocol. After six days of ovulation, the cows were slaughtered and the ovaries and CL were collected. The CLs were processed and photomicrographs were taken under light microscopy to assess the vascular volume density (Vv) by stereology, and scanning electron microscopy (SEM) was used to perform ultrastructural analysis of the microvasculature. The Vv in stimulated and superovulated cows significantly increased (P ≤ 0.0001) when compared to control, indicating that the eCG is able to induce angiogenic activity in bovine CL. However, no significant differences were observed between stimulated and superovulated cows. The SEM demonstrated ratings indicative of angiogenesis, marked by several button-shaped projections in the capillaries, and the presence of more dilated capillaries in CL treated with eCG. These morphological findings are evidence of an angiogenic effect of the eCG treatment in CL of cows., (© 2015 Wiley Periodicals, Inc.)

The objective of the experiment was to compare the use of a PGF2α analogue (Cloprostenol) IM, with an intravaginal progestagen sponge, flurogestone acetate (FGA), and equine chorionic gonadotropin (eCG) IM application protocol. A total of 30 cyclical hair ewes (54.07 ± 0.5 kg live weight, body condition score 3.5 ± 0.5, and age 3 ± 1 years) were used. For the control group ewes (n = 15), intravaginal sponges (IS) impregnated with 20 mg of FGA were inserted for 12 days with 500 IU of eCG IM at sponges withdrawal. For the PG group ewes (Treatment group n = 15), two injections of Cloprostenol (75 mcg) were given 12 days apart. The presence of estrus was detected using two rams with 8 h interval beginning at the end of the treatment. Progesterone concentrations in blood were measured by solid phase radioimmunoassay. A student's t test was performed to analyze the duration of estrus and the interval between the ends of the treatment and the onset of estrus (ET-OE) presentation. Progesterone levels were compared with two-way ANOVA, with treatment, and day of menstrual cycle as fixed factors. Treatment costs ratio was calculated by dividing the total costs of FGA IS application between total costs of Cloprostenol application. Significant differences (P < 0.05) were found in the (ET-OE) interval and estrus duration. For the control group, estrus was presented at 30 + 8.2 h; in treatment group, at 44 h after the last application, duration of estrus was 54.9 + 8.34 h, and 41 + 1.83 h for the control and treatment group, pregnancy rates were 53.3 and 60.0 %, respectively. Significant differences (P < 0.001) were found from days 9 to 13 on Progesterone levels in both treatments. Treatment costs of Cloprostenol protocol were 2.63 cheaper than FGA including disposable material, biological products, and labor. It was concluded that Cloprostenol could be an effective tool in estrus synchronization in hair sheep in tropical areas.

Controlled ovarian hyperstimulation is commonly used in fertility treatment. Evidence suggests that this could alter the endometrial environment and influence implantation rate. However, the mechanisms underlying this disruption are unknown. A recently developed rat ovarian hyperstimulation (OH) model found alterations in the localisation and expression of several molecules associated with implantation, as well as an increase in luminal fluid at the time of implantation. The present study investigated the effects of OH in rats on the expression of fluid-transporting molecules aquaporin 5 (AQP5) and claudin 4. The expression of these proteins was investigated in uterine luminal epithelial cells of rats undergoing OH and compared with normal pregnancy. There was a significant increase in AQP5 protein in OH rats at the time of implantation, along with a loss of the mesometrial staining gradient, which is thought to contribute to implantation position. At the same time, there was a significant decrease in claudin 4 protein. These results suggest that OH in rats causes a dysregulation in uterine fluid dynamics through modifications to fluid-transporting molecules, resulting in an unfavourable implantation environment for the blastocyst.

The uterus plays an essential role in mammalian reproduction and is a target of several hormonal protocols used to improve fertility in cattle. Many studies highlighted the importance of eCG treatment following fixed-time artificial insemination in improving follicular growth, ovulation and pregnancy rates in cattle. Moreover, eCG has been implicated in angiogenesis, leading to important changes in uterine blood flow and vascularisation. However, there is still a lack of information regarding the specific alterations induced by eCG upon glandular and vascular characteristics of bovine uterus. To investigate the influence of eCG on: uterine thickness and area; uterine artery diameter and area; uterine vascular and gland density; and the expression of the VEGFA-system, the uteri of crossbred beef cows were collected. All cows were submitted to follicular wave emergence synchronization. On day four of protocol, cows submitted to superovulation (n = 6) received 2000 IU eCG, on day eight, after expected follicular deviation, cows submitted to stimulatory treatment (n = 5) received 400 IU eCG. Control cows (n = 5) did not receive eCG. On day five po cows were subjected to ultrassonographic evaluation and slaughtered for uterine tissue sampling on day six po. Uterine vessels and glands were quantified by the counting point stereological method. The VEGFA-system was localized in different cellular types, showing no qualitative or quantitative differences in the site of expression or the intensity of the positive signal among the groups. Vascular density was decreased in the endometrium of stimulated and myometrium of superovulated cows compared with the control ones, which showed higher vascular density in the myometrium and endometrium of the ipsilateral uterine horn. The uterine gland density was higher in superovulated compared with stimulated and control cows. Thus, we can infer that stimulatory or superovulatory treatments with eCG influence the vascular density in the endometrium and myometrium in cattle., (© 2014 Blackwell Verlag GmbH.)

This study aims to develop at different seasons, for local North African Maure goats, synchronizing protocols simultaneously to the standard 'S' protocol using progestagens in association with prostaglandins and gonadotropin. In late May, 40 goats were assigned to either the 'S' protocol or to a protocol where oestrus and ovulation were induced by the buck effect in single-injection progesterone-treated goats and provoking early luteolysis using prostaglandin 9 days after exposure to bucks 'B'. During the 72 h after the treatments ended, 15 and 5 goats expressed oestrus in the 'S' and 'B' protocols (p < 0.01). Mean time to oestrus was shorter for 'S' than for 'B' goats. Ovulation rate averaged 2.1 ± 0.22 and 1.60 ± 0.35 for, respectively, 'S' and 'B' goats (p > 0.05). During mid-September, 60 goats were assigned to either 'S' treatment, 'PGF' treatment where oestrus and ovulation were synchronized using two injections of prostaglandin 11 days apart or to 'GnRH' treatment where the goats had their oestrus and ovulation synchronized with a GnRH (day 0)-prostaglandin (day 6)-GnRH (day 9) sequence. More 'S' goats were detected in oestrus over the 96-h period after the end of the treatments (88.8, 73.7 and 55% in 'S', 'PGF' and 'GnRH' treatments, respectively; p < 0.05). Mean ovulation rates were 2.3 ± 0.27, 1.33 ± 0.27 and 1.33 ± 0.27 for, respectively, 'S', 'PGF' and 'GnRH' goats (p < 0.001). Despite a similar ovulatory response to 'S' protocol, efficiency of prostaglandin and GnRH-based treatments should be tested in mid-breeding season., (© 2014 Blackwell Verlag GmbH.)

The objective of this study was to investigate the effects of eCG and temporary calf removal (TCR) associated with progesterone (P4) treatment on the dynamics of follicular growth, CL size, and P4 concentrations in cyclic (n = 36) and anestrous (n = 30) Nelore cows. Cyclic (C) and anestrous (A) cows were divided into three groups. The control group received 2 mg of estradiol benzoate via intramuscular (IM) injection and an intravaginal device containing 1.9 g of P4 on Day 0. On Day 8, the device was removed, and the animals received 12.5 mg of dinoprost tromethamine IM. After 24 hours, the animals received 1 mg of estradiol benzoate IM. In the eCG group, cows received the same treatment described for the control group but also received 400 UI of eCG at the time of device removal. In the TCR group, calves were separated from the cows for 56 hours after device removal. Ultrasound exams were performed every 24 hours after device removal until the time of ovulation and 12 days after ovulation to measure the size of the CL. On the same day as the CL measurement, blood was collected to determine the plasma P4 level. Statistical analyses were performed with a significance level of P ≤ 0.05. In cyclic cows, the presence of the CL at the beginning of protocol resulted in a smaller follicle diameter at the time of device removal (7.4 ± 0.3 mm in cows with CL vs. 8.9 ± 0.4 mm in cows without CL; P = 0.03). All cows ovulated within 72 hours after device removal. Anestrous cows treated with eCG or TCR showed follicle diameter at fixed-timed artificial insemination (A-eCG 10.2 ± 0.3 and A-TCR 10.3 ± 0.5 mm) and follicular growth rate (A-eCG 1.5 ± 0.2 and A-TCR 1.3 ± 0.1 mm/day) similar to cyclic cows (C-eCG 11.0 ± 0.6 and C-TCR 12.0 ± 0.5 mm) and (C-eCG 1.4 ± 0.2 and C-TCR 1.6 ± 0.2 mm/day, respectively; P ≤ 0.05). Despite the similarities in CL size, the average P4 concentration was higher in the A-TCR (9.6 ± 1.4 ng/mL) than in the A-control (4.0 ± 1.0 ng/mL) and C-TCR (4.4 ± 1.0 ng/mL) groups (P < 0.05). From these results, we conclude that eCG treatment and TCR improved the fertility of anestrous cows by providing follicular growth rates and size of dominant follicles similar to cyclic cows. Additionally, TCR increases the plasma concentrations of P4 in anestrous cows., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Objective: To investigate the effects of V1A receptor antagonist through inhibition of vasopressin-induced VEGF secretion in an experimental model., Study Design: Thirty rats were randomly divided into five groups. Four groups were given 10IU pregnant mare serum gonadotropin/day (sc) at 8:00-8:30am on days 22-25 of life. They were administered 30IU hCG at 8:00-8:30am on day 26 of life. On days 26 and 27 of life at 8:00am and 4:00pm, (ip) per animal, 50μg/kg/day GnRH antagonist in the GnRH antagonist group, 0.3mg relcovaptan in the high dose relcovaptan group, and 0.15mg relcovaptan in the low dose relcovaptan group were administered. The control group was given the same dosage of 0.9% saline solution (ip) on days 22-26 day of life. The main outcomes were weight gain, ovarian weights, peritoneal fluid VEGF values, corpus luteum count, and atretic follicle count., Results: Weight gain was highest in the OHSS group; it was almost twice as much in the OHSS group than it was in the control group. Ovarian weights were significantly lower in all treatment groups (p=0.03). There was no statistically significant difference in ovarian weights between the GnRH antagonist and relcovaptan groups (p=0.176). The evaluation of peritoneal fluid VEGF-A levels revealed statistically significant differences between levels in the treatment groups and in the OHSS group (p=0.005). Atretic follicle count in the OHSS group was significantly lower (p=0.048). In all treatment groups, CL counts were prominently lower than they were in the OHSS group (p=0.002)., Conclusion: Relcovaptan may be a novel strategy for decreasing risk of OHSS by inhibition of vasopressin-induced VEGF secretion through V1A receptor antagonist., (Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.)