RESUMO
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 × 106 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.
Assuntos
Gonadotropina Coriônica , Sincronização do Estro , Hormônio Liberador de Gonadotropina , Inseminação Artificial , Animais , Feminino , Inseminação Artificial/veterinária , Hormônio Liberador de Gonadotropina/farmacologia , Hormônio Liberador de Gonadotropina/administração & dosagem , Ovinos/fisiologia , Gonadotropina Coriônica/farmacologia , Gonadotropina Coriônica/administração & dosagem , Gravidez , Sincronização do Estro/métodos , Progesterona/sangue , Progesterona/farmacologia , Progesterona/administração & dosagem , Estações do Ano , Acetato de Medroxiprogesterona/administração & dosagem , Acetato de Medroxiprogesterona/farmacologia , Ovulação/efeitos dos fármacos , Ovulação/fisiologia , Gonadotropinas Equinas/farmacologia , Gonadotropinas Equinas/administração & dosagemRESUMO
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.
Assuntos
Inseminação Artificial , Animais , Bovinos , Feminino , Inseminação Artificial/veterinária , Inseminação Artificial/métodos , Gravidez , Proteínas Recombinantes/farmacologia , Proteínas Recombinantes/administração & dosagem , Gonadotropina Coriônica/farmacologia , Gonadotropina Coriônica/administração & dosagem , Sincronização do Estro/métodos , Progesterona/farmacologia , Progesterona/administração & dosagem , Gonadotropinas Equinas/farmacologia , Gonadotropinas Equinas/administração & dosagem , Taxa de GravidezRESUMO
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.
Assuntos
Gonadotropinas Equinas , Inseminação Artificial , Ovulação , Animais , Feminino , Gravidez , Bovinos/fisiologia , Inseminação Artificial/veterinária , Ovulação/efeitos dos fármacos , Gonadotropinas Equinas/farmacologia , Gonadotropinas Equinas/administração & dosagem , Gonadotropina Coriônica/farmacologia , Gonadotropina Coriônica/administração & dosagem , Aborto Animal , Gravidez de Gêmeos , Progesterona/administração & dosagem , Progesterona/farmacologia , Taxa de GravidezRESUMO
Equine chorionic gonadotropin (eCG) is a glycoprotein hormone widely used in timed artificial ovulation (TAI) and superovulation protocols to improve the reproductive performance in livestock. Until recently, the only eCG products available in the market for veterinary use consisted in partially purified preparations of pregnant mare serum gonadotropin (PMSG). Here, a bioactive recombinant eCG (reCG) produced in suspension CHO-K1 cells was purified employing different chromatographic methods (hydrophobic interaction chromatography and reverse-phase (RP)-HPLC) and compared with a RP-HPLC-purified PMSG. To gain insight into the structural and functional characteristics of reCG, a bioinformatics analysis was performed. An exhaustive characterization comprising the determination of the purity degree, aggregates and nicked forms through SDS-PAGE, RP-HPLC and SEC-HPLC was performed. Higher order structures were studied by fluorescence spectroscopy and SEC-HPLC. Isoforms profile were analyzed by isoelectric focusing. Glycosylation analysis was performed through pulsed amperometric detection and PNGase F treatment following SDS-PAGE and weak anion exchange-HPLC. Slight differences between the purified recombinant hormones were found. However, recombinant molecules and PMSG exhibited variations in the glycosylation pattern. In fact, differences in sialic acid content between two commercial preparations of PMSG were also obtained, which could lead to differences in their biological potency. These results show the importance of having a standardized production process, as occurs in a recombinant protein bioprocess. Besides, our results reflect the importance of the glycan moieties on eCG conformation and hence in its biological activity, preventing denaturing processes such as aggregation.
Assuntos
Gonadotropina Coriônica , Gonadotropinas Equinas , Gravidez , Feminino , Animais , Cavalos , Glicosilação , Proteínas Recombinantes/química , Eletroforese em Gel de PoliacrilamidaRESUMO
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.
Assuntos
Gonadotropina Coriônica , Transferência Embrionária , Gravidez , Humanos , Feminino , Bovinos , Cavalos , Animais , Gonadotropina Coriônica/farmacologia , Transferência Embrionária/veterinária , Gonadotropinas Equinas/farmacologia , Progesterona , Corpo Lúteo , Estradiol/farmacologia , Inseminação Artificial/veterinária , Inseminação Artificial/métodos , Sincronização do Estro/métodos , Ensaios Clínicos Veterinários como AssuntoRESUMO
We aimed to evaluate the morphological ovarian response to equine chorionic gonadotropin (eCG) prior to ovum pick-up (OPU) and its effects on the molecular phenotype of immature cumulus-oocyte complexes (COCs) from Nelore cow (Bos indicus) donors. To this end, 20 Nelore cows were distributed randomly into the synchronized-OPU (Sync-OPU) and synchronized plus stimulated-OPU (Sync + eCG-OPU) groups using a cross-over experimental design, as each cow was used in both treatments. On a random day of the estrus cycle (Day 0), all cows received an intravaginal implant with 1.0 g of progesterone and 2 mg IM of estradiol benzoate. On the morning of Day 3, only the Sync + eCG-OPU group received 400 IU of eCG IM. On the morning of Day 5, the P4 device was removed and OPU was conducted in both groups. Before OPU management, ultrasonography was used to identify and measure the follicles. The aspirated COCs were morphologically classified based on their cumulus cells (CC) layers and the texture of the ooplasm. The COCs classified as Grade 1, Grade 2, and Grade 3 were considered viable and used for the assessment of quality markers. Oocytes and CC were mechanically separated from pools of 25 immature COCs of the Sync-OPU and Sync + eCG-OPU groups immediately after the follicular aspiration and stored at -80 °C until RNA extraction. Relative quantification of several markers for oocyte quality was assessed by RT-qPCR. The eCG treatment increased the number of follicles sized 3.0-5.0 mm and >5.0 mm compared to that in Sync-OPU group. Moreover, the protocol with eCG improved the total number of oocytes and the number of viable oocytes, which is related to a high number of oocytes in Grade 3. Regarding the impact on transcriptional regulation in immature oocytes, the mRNA encoding BMP15, SMAD1, SMAD2, SMAD3, ACACA, and CPT1A was upregulated in Sync + eCG-OPU compared with the Sync-OPU group. Moreover, the relative mRNA abundance of CTSZ, a member of the cathepsins family functionally related to reduced oocyte competence, was lower in the Sync + eCG-OPU group than in the Sync-OPU group. In addition, CC CTSB, CTSS, and CTSK mRNA abundances were lower in the Sync + eCG-OPU group than in the Sync-OPU group. However, the relative abundance of AREG and EREG mRNA was higher in CC recovered from cows stimulated with eCG. In conclusion, the eCG approach addressing follicular stimulation in Nelore cows had a positive impact on early antral follicle development, followed by a positive morphological and molecular phenotype in bovine COCs.
Assuntos
Gonadotropinas Equinas , Recuperação de Oócitos , Animais , Bovinos , Gonadotropina Coriônica/farmacologia , Feminino , Expressão Gênica , Gonadotropinas Equinas/farmacologia , Cavalos , Recuperação de Oócitos/veterinária , OócitosRESUMO
Equine chorionic gonadotropin (eCG) is a heterodimeric glycoprotein hormone produced by pregnant mares that has been used to improve reproductive performance in different domestic species. Several strategies to produce the hormone in a recombinant way have been reported; nevertheless, no approach has been able to produce a recombinant eCG (reCG) with significant in vivo bioactivity or in sufficient quantities for commercial purposes. For this reason, the only current product available on the market consists of partially purified preparations from serum of pregnant mares (PMSG). Herein, we describe a highly efficient process based on third-generation lentiviral vectors as delivery method for the production of reCG in suspension CHO-K1 cells, with productivities above 20 IU 106 cell-1.d-1 and 70% purification yields after one purification step. Importantly, reCG demonstrated biological activity in cattle, since around 30 µg of reCG were needed to exert the same biologic effect of 400 IU of PMSG in an ovulation synchronization protocol. The results obtained demonstrate that the developed strategy represents an attractive option for the production of reCG and constitutes an auspicious alternative for the replacement of animals as a source of PMSG.
Assuntos
Gonadotropina Coriônica , Gonadotropinas Equinas , Animais , Células CHO , Bovinos , Gonadotropina Coriônica/farmacologia , Cricetinae , Cricetulus , Feminino , Gonadotropinas Equinas/farmacologia , Cavalos , Ovulação , GravidezRESUMO
The aim of this study was to evaluate the effect of equine chorionic gonadotropin (eCG) at the end of progesterone (P4) treatment on follicular and luteal characteristics during transition period (TP) and reproductive breeding season (RP). A total of 13 crossbred mares were distributed in two experimental groups in the spring and summer (n = 26). The animals received intravaginal P4 (1.9 g) releasing device from D0 to D10. On removal of P4 device, the mares received 400 IU of eCG (eCG group) or saline solution (control group). Human chorionic gonadotropin (hCG; 1.750 IU) was administered (DhCG) as soon as ovulatory follicle (OF) ≥35 mm was detected. Ovarian ultrasonography was performed from D0 until 15 days after ovulation. Blood samples were collected on D0, D5, D10, DhCG, 9 days after ovulation (CL9D), and 13 days after ovulation (CL13D). P4 and estradiol concentrations were assessed by chemiluminescence. Data were compared by Tukey test at P < .05. Ovulation rate was similar (P = .096) between seasons (RP = 100%; TP = 70%) but occurred earlier (P = .015) in RP (34.8 ± 10.1 hours) compared with TP (42.0 ± 10.4 hours). Interactions between season and treatment were observed for OF diameter (mm) (RP/control = 36.2 ± 1.8ab; RP/eCG = 32.9 ± 2.8 b; TP/control = 32.2 ± 1.2 b; TP/eCG = 37.2 ± 1.9a; P = .004) and for corpus luteum (CL) diameter (mm) on CL13D (RP/control = 25.4 ± 3.5a; RP/eCG = 22.5 ± 1.8ab; TP/control = 21.6 ± 4.9 b; TP/eCG = 27.4 ± 4.3a; P = .023), although no differences were observed for serum P4 on CL13D (RP/control = 6.0 ± 3.1 ng/mL; RP/eCG = 5.8 ± 0.9 ng/mL; TP/control = 3.6 ± 2.7 ng/mL; TP/eCG = 5.1 ± 2.3 ng/mL; P = .429) or for day of structural CL regression (RP/control = 12.8 ± 1.9; RP/eCG = 12.1 ± 1.1; TP/control = 11.0 ± 1.7; TP/eCG = 13.2 ± 2.0; P = .102). The application of eCG at the moment of P4 implant removal seemed to increase the capacity of luteal maintenance during spring TP. However, eCG treatment was worthless during the breeding season.
Assuntos
Gonadotropinas Equinas , Inseminação Artificial , Animais , Gonadotropina Coriônica/farmacologia , Corpo Lúteo , Feminino , Gonadotropinas Equinas/farmacologia , Cavalos , Inseminação Artificial/veterinária , Indução da Ovulação/veterináriaRESUMO
The present study evaluated the effect of the aqueous extract from leaves of E. speciosa on some physiological and biochemical parameters of reproduction and the onset of puberty in pregnant mare serum gonadotropin (PMSG)-primed immature female rats. High pressure liquid chromatography (HPLC) was used to quantify the phenolic compounds in the methanol/methylene chloride (1:1) extract, the ethanolic and ethyl acetate fractions and the aqueous residue of E. speciosa. E. speciosa (0, 8, 32 or 64 mg/kg) were administered for 15 days to 24 non-PMSG-primed and 24 primed rats with 0.01 IU of PMSG. At the end of the treatment period, animal were sacrificed and their body, ovarian, uterine weight, ovarian protein or cholesterol level, as well as data on puberty onset were recorded. Of the 16 polyphenolic compounds quantitatively revealed in the extracts and fractions of E. speciosa after HPLC analysis, quercetin, rutin, apigenin and eugenol were the most abundant. Non-primed rats showed a significant increase (P < 0.05) in the uterine relative weight at the dose of 8 mg/kg when compared with the other treatments. The uterine proteins and the ovarian cholesterol (P < 0.05), respectively, showed a reduction at doses of 64 mg/kg and 32 mg/kg in non-primed rats. However in PMSG-primed rats, a significant decrease (P < 0.05) was observed in ovarian cholesterol at 64 mg/kg. In conclusion, E. speciosa potentializes the PMSG-inducing effect on folliculogenesis in PMSG-primed rats.
Assuntos
Maturidade Sexual , Animais , Feminino , Gonadotropinas Equinas , Cavalos , Ovário , Gravidez , Ratos , Reprodução , ÚteroRESUMO
We tested FSHp, eCG and FSHp + eCG to establish ovum pick-up (OPU) and in vitro maturation method in spotted paca. Eight healthy adult females were subjected to each of four treatments to stimulate ovarian follicular growth. All females were subjected to a hormonal protocol using a single dose of 45 mg of injectable progesterone and single intramuscular injection of 0.075 mg d-cloprostenol on day 6. Ovarian stimulation was carried out as follows: in Group TFE (FSHp and eCG), animals were treated with a single dose of 80 mg of FSHp and 200 IU of eCG intramuscularly on day 6 after the application of progesterone; in Group TF (FSHp), they were treated with a single dose of 80 mg of FSHp intramuscularly on day 6 after application of progesterone; in Group treatment eCG, they were treated with 200 IU of eCG intramuscularly on day 6 after application of progesterone; and in Group TC (saline solution), 1 ml of saline solution was administered to control does. The OPU was performed between 22 and 26 hr after gonadotropin treatments. All recovered oocytes were placed into maturation media and incubated for 24 hr. There were no differences among the mean number of observed follicles, aspirated follicles and oocytes recovered per treatment. Oocyte maturation rates did not differ among groups, except, TF and treatment eCG oocytes had greater maturation rates than TC oocytes. In this study, gonadotropin administration failed to superovulate treated does and increase oocyte retrieval efficiency. Despite the feasibility of the procedure, further studies are needed to develop and refine hormonal protocols for oocyte recovery and in vitro maturation in this species.
Assuntos
Cuniculidae/fisiologia , Técnicas de Maturação in Vitro de Oócitos/veterinária , Recuperação de Oócitos/veterinária , Animais , Cloprostenol/farmacologia , Feminino , Hormônio Foliculoestimulante/farmacologia , Gonadotropinas Equinas/farmacologia , Técnicas de Maturação in Vitro de Oócitos/métodos , Recuperação de Oócitos/métodos , Folículo Ovariano/efeitos dos fármacos , Indução da Ovulação/métodos , Indução da Ovulação/veterinária , Progesterona/farmacologiaRESUMO
Background: Lacaune is an important sheep breed and shows reproductive seasonality which affects dairy product supply. The most common hormonal protocols are based on progesterone-releasing devices in order to reach reproductive activity. Due to international consumer demand for natural, green, and clean methods, such issues encourage the minimal or absent use of synthetic hormones in livestock. In this sense, the present study tested the efficiency of induction and estrus synchronization protocols in lacaune ewes, considering: application interval of synthetic prostaglandin at the breeding season; length of synthetic progesterone exposure; and eCG dose at the outbreeding season. Materials, Methods & Results: In study 1: 52 ewes were submitted to estrus synchronization protocol using two doses of d-cloprostenol 7 or 9 days apart. In study 2: 47 ewes were submitted to estrus induction protocol using an intravaginal device containing 60 mg of medroxyprogesterone acetate maintained for 6, 9, or 12 days. In study 3: 46 ewes were submitted to synchronized estrus induction protocol using intravaginal progesterone device for six days. On the fifth day, d-cloprostenol and 300 IU or 400 IU of eCG were administered. For all studies, signs of estrus beginning/ending were recorded and natural mating was performed. After 30 days, transrectal ultrasound was performed for pregnancy diagnosis. In study 1, groups presented similar rate of estrus behavior (78.8%) and pregnancy (average 75.6%). Meanwhile, compared to G9days (37.8 ± 7.2 h), interval between the second dose of prostaglandin and beginning of estrus was smaller in G7days (31.5 ± 7.8 h). No difference was found among variables studied in study 2 (P > 0.05), nevertheless the SD for the estrus duration was shorter (P < 0.05) in G6days and G12 days when compared with the G9days . Equally, the SD...
Assuntos
Feminino , Animais , Gonadotropinas Equinas/administração & dosagem , Ovinos/fisiologia , Progesterona/administração & dosagem , Prostaglandinas Sintéticas/administração & dosagem , Sincronização do Estro/métodos , Endogamia , Técnicas de Reprodução Assistida/veterináriaRESUMO
Equine chorionic gonadotrophin (eCG) is a hormone having FSH/LH effects. It can be used to enhance sperm quality in male goats (bucks) during the non-breeding season. In a previous study carried out during the non-breeding season, we treated ten bucks with eCG (leaving nine untreated animals as control). Over a 20-day period, the treated bucks received an initial dose of 800 IU of eCG, followed by four doses of 500 IU. We found eCG enhanced semen quality, however, as also happens in female goats (does), eCG also induced a high titer of anti-eCG antibodies. In does, this lowers fertility. The aim of the present study was to determine if the eCG treatment carried out on bucks during the non-breeding season had any negative effects on their reproductive status during the following breeding season. We measured serum concentration of testosterone and anti-eCG antibody, as well as key testicular and seminal characteristics. This study commenced 91 days after the final dose of eCG in the previous study. The anti-eCG titer was higher in the treatment bucks than in untreated ones (181.7 ± 61.3 ng/µL vs 31.1 ± 10.7 ng/µL; P < 0.05). However, there were no significant differences between treated and untreated bucks in testosterone concentration, scrotal circumference, testes pixel intensity, fresh and thawed semen characteristics, or sperm cryoresistance. So, although the eCG-treated bucks had greater titers of anti-eCG antibodies, their reproductive pattern was unaffected.
Assuntos
Cruzamento , Cabras/fisiologia , Gonadotropinas Equinas/farmacologia , Reprodução , Espermatozoides/efeitos dos fármacos , Animais , Masculino , Estações do Ano , Análise do Sêmen/veterinária , Testosterona/sangueRESUMO
This study evaluated the efficiency application of low dose of equine chorionic gonadotropin (eCG) using Hou Hai acupoint for fixed-term artificial insemination (FTAI) in beef cows. Seventy cows received intravaginal devices with progesterone, and 3 mg of estradiol benzoate on day zero (D0) of FTAI. On D9, the devices were removed, and 150 μg of prostaglandin F2α was applied; the animals were then randomly distributed to three treatment groups: T1 (n = 23), 300 IU of eCG intramuscularly (IM); T2 (n = 23), 90 IU of eCG at acupoint Hou Hai; and T3 (n = 25), 90 IU of eCG at false acupoint IM. On D10, the animals received 1 mg of gonadotropin-releasing hormone IM, and FTAI was performed, 52 h after the devices were removed. The quantitative variables with normal distribution were assessed using analysis of variance at 5% probability. For variables that did not show normal distribution, Kruskal-Wallis test at 5% probability was used. No significant difference was noted among the groups (P > 0.05) for the interval between device withdrawal at ovulation (58.2 ± 12.2 h), diameter of the largest follicle on D9 (9.9 ± 2.2 mm), diameters of the ovulatory follicle (12.2 ± 3.0 mm) and the second largest follicle (6.7 ± 2.1 mm), follicular growth rate (0.8 ± 0.3 mm/d), ovulation rate (82%), corpus luteum size (2.32 ± 0.35 cm2), and pregnancy rate (58.67%). The protocol cost per animal was US$ 10.67 (T1) and US$ 8.50 (T2 and T3). The use of 90 IU of eCG applied at Hou Hai acupoint or at false acupoint caused satisfactory synchronization of estrus in beef cows. In addition, this procedure was cost-effective.(AU)
Avaliou-se a eficiência da aplicação de subdose gonadotrofina coriônica equina (eCG) utulizando como via de administração o acuponto Hou Hai em protocolos de inseminação artificial em tempo fixo (IATF) para vacas de corte. Setenta vacas receberam no dia zero (D0) do protocolo dispositivos intravaginais com progesterona, e 3mg de benzoato de estradiol. No D9 foram retirados os dispositivos, aplicado 150µg de prostaglandina F2α e distribuídos os animais aleatoriamente em três tratamentos: T1 (n=23) - 300UI de eCG por via intra muscular (IM); T2 (n=23) - 90UI de eCG no acuponto Hou Hai e T3 (n=25) - 90UI de eCG em falso acuponto, IM. No D10 os animais receberam 1mg de GnRH por via IM e foi realizada a IATF, 52 horas após a retirada dos dispositivos. Para as variáveis quantitativas com distribuição normal foi utilizada ANOVA, a 5% de probabilidade. Para as variáveis que não apresentaram distribuição normal, utilizou-se o teste de Kruskal-Wallis, a 5% de probabilidade. Não houve diferença (P >0,05) para o intervalo entre a retirada do dispositivo à ovulação (58,2 ± 12,2h), diâmetro do maior folículo no D9 (9,9 ± 2,2mm), diâmetros do folículo ovulatório (12,2 ± 3,0mm) e do segundo maior folículo (6,7 ± 2,1mm), taxa de crescimento folicular (0,8 ± 0,3mm/d), taxa de ovulação (82%), área de corpo lúteo (2,32 ± 0,35 cm2) e taxa de prenhez (58,67%). O custo do protocolo por animal foi de US$ 10,67 (T1) e US$ 8,50 (T2 e T3). Sugere-se que a utilização de 90UI de eCG aplicada no acuponto Hou Hai ou em falso acuponto foram satisfatórios na sincronização de estro de vacas de corte. Além disso, possibilita a redução do custo do protocolo.(AU)
Assuntos
Animais , Feminino , Bovinos , Gonadotropina Coriônica/administração & dosagem , Gonadotropinas Equinas/administração & dosagem , Inseminação Artificial/veterinária , Sincronização do EstroRESUMO
Avaliaram-se diferentes momentos de aplicação da gonadotrofina coriônica equina (eCG) em protocolos de inseminação artificial em tempo fixo (IATF) para vacas de leite. Foram utilizadas 76 fêmeas, as quais receberam, no dia zero (D0) do protocolo, dispositivos intravaginais de progesterona, sendo esses retirados no D9, e os animais foram, então, distribuídos aleatoriamente em três tratamentos: T1 - aplicação de eCG no momento da retirada dos dispositivos; T2 e T3 - aplicação de eCG 48h e 24h antes da retirada dos dispositivos, respectivamente. No D10 os animais receberam 1mg de GnRH, e a IATF foi realizada 52 horas após a retirada do implante. Não houve diferença (P>0,05) para intervalo entre a retirada do implante à ovulação (72,56±3,92h), o diâmetro do maior folículo no D9 (10,88±1,49mm), o diâmetro do folículo ovulatório (15,15±1,16mm) e do segundo maior folículo (7,49±0,52mm), a taxa de crescimento folicular (1,38±0,04mm/dia), a taxa de ovulação (96,67%), o intervalo entre diâmetro final e inicial do folículo dominante (73,49±3,84h), a área de corpo lúteo (2,27±0,43cm²), a porcentagem de CL no ovário direito (53,00%) e no esquerdo (26,33%) e a taxa de gestação (33,33%). O momento da aplicação da eCG não influenciou na eficiência do protocolo. Recomenda-se a utilização da eCG no momento da retirada do implante por otimização do manejo.(AU)
Different times of application of equine chorionic gonadotropin (eCG) in Fixed-Time Artificial Insemination (FTAI) protocols for dairy cows were evaluated. A total of 76 females were used, which received intravaginal progesterone devices on day zero (D0) of the protocol, which were withdrawn on D9, and the animals were then randomly distributed in three treatments: T1- application of eCG at the time of device withdrawal; T2 and T3 - application of eCG 48h and 24h before withdrawal of the devices, respectively. On D10 the animals received 1mg of GnRH and the FTAI was carried out 52 hours after the removal of the implant. There was no difference (P>0.05) for interval from implant removal to ovulation (72.56±3.92h), diameter of the largest follicle of D9 (10.88±1.49mm), diameters of ovulatory follicle (15.15±1.16mm) and of the second largest follicle (7.49±0.52mm), follicle growth rate (1.38±0.04mm/day), ovulation rate (96.67%), interval between final diameter and initial diameter of dominant follicle (73.49±3.84h), corpus luteum area (2.27±0.43cm²), percentage of CL in right ovary (53.00%) and in left ovary (26.33%) and pregnancy rate (33.33%). The time of the application of eCG did not influence the efficiency of the protocol. We recommend the use of eCG at the time of the removal of the implant for management optimization.(AU)
Assuntos
Animais , Feminino , Bovinos , Ovulação , Inseminação Artificial/veterinária , Gonadotropinas Equinas/administração & dosagemRESUMO
This study evaluated the efficiency application of low dose of equine chorionic gonadotropin (eCG) using Hou Hai acupoint for fixed-term artificial insemination (FTAI) in beef cows. Seventy cows received intravaginal devices with progesterone, and 3 mg of estradiol benzoate on day zero (D0) of FTAI. On D9, the devices were removed, and 150 μg of prostaglandin F2α was applied; the animals were then randomly distributed to three treatment groups: T1 (n = 23), 300 IU of eCG intramuscularly (IM); T2 (n = 23), 90 IU of eCG at acupoint Hou Hai; and T3 (n = 25), 90 IU of eCG at false acupoint IM. On D10, the animals received 1 mg of gonadotropin-releasing hormone IM, and FTAI was performed, 52 h after the devices were removed. The quantitative variables with normal distribution were assessed using analysis of variance at 5% probability. For variables that did not show normal distribution, Kruskal-Wallis test at 5% probability was used. No significant difference was noted among the groups (P > 0.05) for the interval between device withdrawal at ovulation (58.2 ± 12.2 h), diameter of the largest follicle on D9 (9.9 ± 2.2 mm), diameters of the ovulatory follicle (12.2 ± 3.0 mm) and the second largest follicle (6.7 ± 2.1 mm), follicular growth rate (0.8 ± 0.3 mm/d), ovulation rate (82%), corpus luteum size (2.32 ± 0.35 cm2), and pregnancy rate (58.67%). The protocol cost per animal was US$ 10.67 (T1) and US$ 8.50 (T2 and T3). The use of 90 IU of eCG applied at Hou Hai acupoint or at false acupoint caused satisfactory synchronization of estrus in beef cows. In addition, this procedure was cost-effective.
Avaliou-se a eficiência da aplicação de subdose gonadotrofina coriônica equina (eCG) utulizando como via de administração o acuponto Hou Hai em protocolos de inseminação artificial em tempo fixo (IATF) para vacas de corte. Setenta vacas receberam no dia zero (D0) do protocolo dispositivos intravaginais com progesterona, e 3mg de benzoato de estradiol. No D9 foram retirados os dispositivos, aplicado 150µg de prostaglandina F2α e distribuídos os animais aleatoriamente em três tratamentos: T1 (n=23) - 300UI de eCG por via intra muscular (IM); T2 (n=23) - 90UI de eCG no acuponto Hou Hai e T3 (n=25) - 90UI de eCG em falso acuponto, IM. No D10 os animais receberam 1mg de GnRH por via IM e foi realizada a IATF, 52 horas após a retirada dos dispositivos. Para as variáveis quantitativas com distribuição normal foi utilizada ANOVA, a 5% de probabilidade. Para as variáveis que não apresentaram distribuição normal, utilizou-se o teste de Kruskal-Wallis, a 5% de probabilidade. Não houve diferença (P >0,05) para o intervalo entre a retirada do dispositivo à ovulação (58,2 ± 12,2h), diâmetro do maior folículo no D9 (9,9 ± 2,2mm), diâmetros do folículo ovulatório (12,2 ± 3,0mm) e do segundo maior folículo (6,7 ± 2,1mm), taxa de crescimento folicular (0,8 ± 0,3mm/d), taxa de ovulação (82%), área de corpo lúteo (2,32 ± 0,35 cm2) e taxa de prenhez (58,67%). O custo do protocolo por animal foi de US$ 10,67 (T1) e US$ 8,50 (T2 e T3). Sugere-se que a utilização de 90UI de eCG aplicada no acuponto Hou Hai ou em falso acuponto foram satisfatórios na sincronização de estro de vacas de corte. Além disso, possibilita a redução do custo do protocolo.
Assuntos
Feminino , Animais , Bovinos , Gonadotropina Coriônica/administração & dosagem , Gonadotropinas Equinas/administração & dosagem , Inseminação Artificial/veterinária , Sincronização do EstroRESUMO
Avaliaram-se diferentes momentos de aplicação da gonadotrofina coriônica equina (eCG) em protocolos de inseminação artificial em tempo fixo (IATF) para vacas de leite. Foram utilizadas 76 fêmeas, as quais receberam, no dia zero (D0) do protocolo, dispositivos intravaginais de progesterona, sendo esses retirados no D9, e os animais foram, então, distribuídos aleatoriamente em três tratamentos: T1 - aplicação de eCG no momento da retirada dos dispositivos; T2 e T3 - aplicação de eCG 48h e 24h antes da retirada dos dispositivos, respectivamente. No D10 os animais receberam 1mg de GnRH, e a IATF foi realizada 52 horas após a retirada do implante. Não houve diferença (P>0,05) para intervalo entre a retirada do implante à ovulação (72,56±3,92h), o diâmetro do maior folículo no D9 (10,88±1,49mm), o diâmetro do folículo ovulatório (15,15±1,16mm) e do segundo maior folículo (7,49±0,52mm), a taxa de crescimento folicular (1,38±0,04mm/dia), a taxa de ovulação (96,67%), o intervalo entre diâmetro final e inicial do folículo dominante (73,49±3,84h), a área de corpo lúteo (2,27±0,43cm²), a porcentagem de CL no ovário direito (53,00%) e no esquerdo (26,33%) e a taxa de gestação (33,33%). O momento da aplicação da eCG não influenciou na eficiência do protocolo. Recomenda-se a utilização da eCG no momento da retirada do implante por otimização do manejo.(AU)
Different times of application of equine chorionic gonadotropin (eCG) in Fixed-Time Artificial Insemination (FTAI) protocols for dairy cows were evaluated. A total of 76 females were used, which received intravaginal progesterone devices on day zero (D0) of the protocol, which were withdrawn on D9, and the animals were then randomly distributed in three treatments: T1- application of eCG at the time of device withdrawal; T2 and T3 - application of eCG 48h and 24h before withdrawal of the devices, respectively. On D10 the animals received 1mg of GnRH and the FTAI was carried out 52 hours after the removal of the implant. There was no difference (P>0.05) for interval from implant removal to ovulation (72.56±3.92h), diameter of the largest follicle of D9 (10.88±1.49mm), diameters of ovulatory follicle (15.15±1.16mm) and of the second largest follicle (7.49±0.52mm), follicle growth rate (1.38±0.04mm/day), ovulation rate (96.67%), interval between final diameter and initial diameter of dominant follicle (73.49±3.84h), corpus luteum area (2.27±0.43cm²), percentage of CL in right ovary (53.00%) and in left ovary (26.33%) and pregnancy rate (33.33%). The time of the application of eCG did not influence the efficiency of the protocol. We recommend the use of eCG at the time of the removal of the implant for management optimization.(AU)
Assuntos
Animais , Feminino , Bovinos , Ovulação , Inseminação Artificial/veterinária , Gonadotropinas Equinas/administração & dosagemRESUMO
The objective of this study was to determine the effect of breed and equine chorionic gonadotropin (eCG) on ovarian response and in vitro embryo production from young goats. Thirty-one (12 Alpine, 10 Nubian, and 9 Saanen) were randomly assigned into three treatments of eCG (T1, 0 IU; T2, 500 IU; and T3, 1000 IU). Alpine goats showed the highest amount and largest size of follicles (P = 0.003). The effect of eCG dose 24 h post application was significant (P < 0.05), and was superior in goats undergoing T2. The aspiration rate of cumulus-oocyte complexes (COC) was 34% (P > 0.05), except for percentage of denuded oocytes, which obtained the highest number (P = 0.003) in the Saanen goats. The same difference was found (P = 0.02) in oocytes grade III in T2 and T3, with 42.5 and 37.9% respectively. In vitro embryo production was 80.0% of IVF/cleavage in the Alpine goats (P = 0.003). Embryo production was the greatest for T2 (69.2%; P = 0.004). T3 goats had higher percentage of morula stage (66.6%; P = 0.030). It is concluded that the application of eCG has a significant effect on the ovarian status, and quality and quantity of embryos with a differential response depending on the breed.
Assuntos
Técnicas de Cultura Embrionária/veterinária , Cabras/fisiologia , Gonadotropinas Equinas/farmacologia , Oócitos/fisiologia , Estações do Ano , Animais , Feminino , Fertilização in vitro , Cabras/embriologia , OvárioRESUMO
Avaliou-se a eficiência da administração de subdoses de eCG nos acupontos Bai Hui e Hou Hai em protocolos de sincronização de estro em cabras. Na primeira etapa, 57 cabras foram distribuídas aleatoriamente em quatro tratamentos: T1- 300UI de eCG intramuscular (IM); T2- 60UI de eCG no acuponto Hou Hai; T3- 60UI de eCG no acuponto Bai Hui e T4- 60UI de eCG IM; e na segunda etapa, 28 cabras foram distribuídas aleatoriamente em três tratamentos: T1- 300UI de eCG IM; T2- 30UI de eCG no acuponto Bai Hui e T3- 30UI de eCG IM. Ao final do tratamento hormonal, as cabras foram monitoradas para detecção do estro, realização das coberturas e avaliação do comportamento reprodutivo. Os dados foram submetidos à análise de normalidade, seguida dos testes estatísticos adequados para cada variável. Na primeira etapa experimental, obteve-se maior duração de estro nas cabras do T1 (P=0,009). Na segunda etapa experimental, obteve-se maior número de animais em estros no T1 (P=0,03). As demais variáveis para ambas as etapas não sofreram influência dos tratamentos (P>0,05), demonstrando que a administração de subdoses de eCG nos acupontos Bai Hui e Hou Hai foi eficiente para sincronizar o estro.(AU)
The efficiency of administration of subdoses of eCG in the Bai Hui and Hou Hai acupoints in oestrus synchronization protocols in goats was evaluated. In the first stage, 57 goats were randomly assigned to four treatments: T1- 300UI of intramuscular eCG (IM); T2- 60UI of eCG in acupoint Hou Hai; T3- 60UI of eCG in the Bai Hui acupoint and T4- 60UI of eCG IM; and in the second stage, 28 goats were randomly assigned to three treatments: T1-300UI of eCG IM; T2-30UI of eCG in the Bai Hui acupoint and T3- 30UI of eCG IM. At the end of the hormonal treatment the goats were monitored for estrus detection, and evaluation of reproductive behavior. The data were submitted to normality analysis, followed by appropriate statistical tests for each variable. In the first experimental stage, a longer duration of estrus in the T1 goats (P= 0.009) was obtained. In the second experimental stage, a greater number of animals were obtained in estrus at T1 (P= 0.03). The other variables for both experiments were not influenced by the treatments (P> 0.05), demonstrating that administration of eCG subdoses in the Bai Hui and Hou Hai acupoints was efficient to synchronize the estrus.(AU)
Assuntos
Animais , Feminino , Cabras , Terapia por Acupuntura/veterinária , Sincronização do Estro/métodos , Gonadotropinas Equinas/administração & dosagem , Ciclo EstralRESUMO
Avaliou-se a eficiência da administração de subdoses de eCG nos acupontos Bai Hui e Hou Hai em protocolos de sincronização de estro em cabras. Na primeira etapa, 57 cabras foram distribuídas aleatoriamente em quatro tratamentos: T1- 300UI de eCG intramuscular (IM); T2- 60UI de eCG no acuponto Hou Hai; T3- 60UI de eCG no acuponto Bai Hui e T4- 60UI de eCG IM; e na segunda etapa, 28 cabras foram distribuídas aleatoriamente em três tratamentos: T1- 300UI de eCG IM; T2- 30UI de eCG no acuponto Bai Hui e T3- 30UI de eCG IM. Ao final do tratamento hormonal, as cabras foram monitoradas para detecção do estro, realização das coberturas e avaliação do comportamento reprodutivo. Os dados foram submetidos à análise de normalidade, seguida dos testes estatísticos adequados para cada variável. Na primeira etapa experimental, obteve-se maior duração de estro nas cabras do T1 (P=0,009). Na segunda etapa experimental, obteve-se maior número de animais em estros no T1 (P=0,03). As demais variáveis para ambas as etapas não sofreram influência dos tratamentos (P>0,05), demonstrando que a administração de subdoses de eCG nos acupontos Bai Hui e Hou Hai foi eficiente para sincronizar o estro.(AU)
The efficiency of administration of subdoses of eCG in the Bai Hui and Hou Hai acupoints in oestrus synchronization protocols in goats was evaluated. In the first stage, 57 goats were randomly assigned to four treatments: T1- 300UI of intramuscular eCG (IM); T2- 60UI of eCG in acupoint Hou Hai; T3- 60UI of eCG in the Bai Hui acupoint and T4- 60UI of eCG IM; and in the second stage, 28 goats were randomly assigned to three treatments: T1-300UI of eCG IM; T2-30UI of eCG in the Bai Hui acupoint and T3- 30UI of eCG IM. At the end of the hormonal treatment the goats were monitored for estrus detection, and evaluation of reproductive behavior. The data were submitted to normality analysis, followed by appropriate statistical tests for each variable. In the first experimental stage, a longer duration of estrus in the T1 goats (P= 0.009) was obtained. In the second experimental stage, a greater number of animals were obtained in estrus at T1 (P= 0.03). The other variables for both experiments were not influenced by the treatments (P> 0.05), demonstrating that administration of eCG subdoses in the Bai Hui and Hou Hai acupoints was efficient to synchronize the estrus.(AU)
Assuntos
Animais , Feminino , Cabras , Terapia por Acupuntura/veterinária , Sincronização do Estro/métodos , Gonadotropinas Equinas/administração & dosagem , Ciclo EstralRESUMO
The objective of this study was to evaluate effects of eCG on vascularization and development of feline ovarian tissue xenografted to immunosuppressed mice. Feline ovarian fragments (â¼1â¯mm3) were transplanted under the renal capsule of 20 adult, ovariectomized, C57BL/6 SCID female mice. At 45â¯d after transplantation, 10 mice (controls) were euthanized and the remainder given 10 IU of eCG (and sacrificed 48â¯h later). Transplants were recovered immediately after death, fixed, sectioned, and stained with periodic acid-Schiff (PAS). Fragment volume (Cavallieri principle) and vascularization were assessed. Mean xenotransplant volume for control and treatment groups was 0.17⯱â¯0.03 and 0.37⯱â¯0.13â¯mm3, respectively (Pâ¯=â¯0.0952); vascular volume density, 30.3⯱â¯11.3 and 49.1⯱â¯8.9% (Pâ¯=â¯0.0281); surface density, 4.1⯱â¯2.4 and 6.2⯱â¯1.7⯵m-1â¯(Pâ¯=â¯0.2222); and vessel total surface, 0.63⯱â¯0.24⯵m2 and 2.28⯱â¯1.05⯵m2â¯(Pâ¯=â¯0.0079). In conclusion, eCG significantly increased vascular volume density of xenotransplanted ovarian tissue and improved its development.