GnRH agonist versus GnRH antagonist in poor ovarian responders: A meta-analysis

The aim of this meta-analysis was to compare the efficacy of gonadotrophin antagonist (GnRH-ant) versus GnRH agonist (GnRHa) as coadjuvant therapy for ovarian stimulation in poor ovarian responders in IVF/intracytoplasmic sperm injection cycles. Search strategies included on-line surveys of databases such as MEDLINE, EMBASE and others. A fixed effects model was used for odds ratio (OR) and effect size (weighted mean difference, WMD). Six trials fulfilled the inclusion criteria (randomized controlled trials). There was no difference between GnRH-ant and GnRHa (long and flare-up protocols) with respect to cycle cancellation rate, number of mature oocytes and clinical pregnancy rate per cycle initiated, per oocyte retrieval and per embryo transfer. When the mete-analysis was applied to the two trials that had used GnRH-ant versus long protocols of GnRHa, a significantly higher number of retrieved oocytes was observed in the GnRH-ant protocols [P = 0.018; WMD: 1.12 (0.18, 2.05)]. However, when the meta-analysis was applied to the four trials that had used GnRH-ant versus flare-up protocols, a significantly higher number of retrieved oocytes (P = 0.032; WMD: -0.51, 95% CI -0.99, -0.04) was observed in the GnRHa protocols. Nevertheless, additional randomized controlled trials with better planning are needed to confirm these results.

Utilização do primeiro ciclo ovulatório da estação reprodutiva para produção de embriões em éguas sob condições tropicais

The efficacy of the first ovulation of the breeding season was determined through the response of first pre-ovulatory follicle of the breeding season to hCG, embryo recovery rate, viability of recovered embryos, serum concentrations of progesterone, and response of first CL to PGF 2α. Thirteenth mares that were in vernal transition were accompanied until the first pre-ovulatory follicle was detected. At this moment, the ovulation was induced with 2,500 IU hCG (IV) and the mares were inseminated every other day until ovulation. Seven days after the ovulation, embryo recovery was performed and the progesterone concentration was determined. After detection of the first pre-ovulatory follicle of breeding season with ≥ 25mm, it took 14.92 ± 10.80 days for the follicle to reach the preovulatory size and 18.00 ± 11.08 days to ovulation. After administration of hCG, 11/13 mares ovulated in 48 hours. These follicles growth 2.19 ± 0.86 mm/day on average. Nine of 13 mares (69.2%) produced embryos and all were considered viable after morfological evaluation and fluorescence exams. The CL appeared competent producing 7.39 ± 2.11 ng/ml P 4 on average, and responding to PGF 2α. According to these results the first ovulatory cycle of the year can be utilized to produce viable embryos.

Recombinant LH supplementation to recombinant FSH during induced ovarian stimulation in the GnRH-antagonist protocol: A meta-analysis

This study aims to compare the efficacy of recombinant LH (rLH) supplementation for ovarian stimulation in gonadotrophin-releasing hormone-antagonist protocol for IVF/intracytoplasmic sperm injection cycles. Search strategies included online surveys of databases. The fixed effects model was used for odds ratio (OR) and effect size (weighted mean difference, WMD). Five trials fulfilled the inclusion criteria. When the meta-analysis was carried out, advantages were observed for the LH supplementation protocol with respect to higher serum oestradiol concentrations on the day of human chorionic gonadotrophin administration (P < 0.0001; WMD: 514, 95% CI 368, 660) and higher number of mature oocytes (P = 0.0098; WMD: 0.88, 95% CI 0.21, 1.54). However, these differences were not observed in the total amount of recombinant FSH (rFSH) administered, days of stimulation, number of oocytes retrieved, the clinical pregnancy rate per oocyte retrieval, the implantation rate and miscarriage rate. This result demonstrates that the association of rLH with rFSH may prevent any decrease in oestradiol after antagonist administration and that a significantly higher number of mature oocytes was available for laboratory work. Nevertheless, it failed to show any statistically significant difference in clinically significant end-points in IVF (implantation and pregnancy rates). Additional randomized controlled trials are needed to confirm these results further. © 2007 Published by Reproductive Healthcare Ltd.

The effect of type of vaginal insert and dose of pLH on embryo production, following fixed-time AI in a progestin-based superstimulatory protocol in Nelore cattle

The objective was to analyze and report field data focusing on the effect of type of progesterone-releasing vaginal insert and dose of pLH on embryo production, following a superstimulatory protocol involving fixed-time artificial insemination (FTAI) in Nelore cattle (Bos taurus indicus). Donor heifers and cows (n = 68; 136 superstimulations over 2 years) received an intravaginal, progesterone-releasing insert (CIDR® or DIB®, with 1.9 or 1.0 g progesterone, respectively) and 3-4 mg of estradiol benzoate (EB) i.m. at random stages of the estrous cycle. Five days later (designated Day 0), cattle were superstimulated with a total of 120-200 mg of pFSH (Folltropin-V®), given twice daily in decreasing doses from Days 0 to 3. All cattle received two luteolytic doses of PGF2α at 08:00 and 20:00 h on Day 2 and progesterone inserts were removed at 20:00 h on Day 3 (36 h after the first PGF2α injection). Ovulation was induced with pLH (Lutropin-V®, 12.5 or 25 mg, i.m.) at 08:00 h on Day 4 with FTAI 12, 24 and in several cases, 36 h later. Embryos were recovered on Days 11 or 12, graded and transferred to synchronous recipients. Overall, the mean (±S.E.M.) number of total ova/embryos (13.3 ± 0.8) and viable embryos (9.4 ± 0.6) and pregnancy rate (43.5%; 528/1213) did not differ among groups, but embryo viability rate (overall, 70.8%) was higher in donors with a DIB (72.3%) than a CIDR (68.3%, P = 0.007). In conclusion, the administration of pLH 12 h after progesterone removal in a progestin-based superstimulatory protocol facilitated fixed-time AI in Nelore donors, with embryo production, embryo viability and pregnancy rates after embryo transfer, comparable to published results where estrus detection and AI was done. Results suggested a possible alternative, which would eliminate the need for estrus detection in donors. © 2006 Elsevier Inc. All rights reserved.

Follicular dynamics and plasma FSH and progesterone concentrations during follicular deviation in the first post-ovulatory wave in Nelore (Bos indicus) heifers

The objective of the present study was to characterize ovarian follicular dynamics and hormone concentrations during follicular deviation in the first wave after ovulation in Nelore (Bos indicus) heifers. Ultrasonographic exams were performed and blood samples were collected every 12 h from the day of estrus until 120-144 h after ovulation in seven females. Deviation was defined as the point at which the growth rate of the dominant follicle became greater than the growth rate of the largest subordinate follicle. Deviation occurred approximately 65 h after ovulation. Growth rate of the dominant follicle increased (P < 0.05) after deviation, while growth rate of the subordinate follicle decreased (P < 0.05). Diameter of the dominant follicle did not differ from the subordinate follicle at deviation (approximately 5.4 mm). The dominant follicle (7.6 mm) was larger (P < 0.05) than the subordinate follicle (5.3 mm) 96 h after ovulation or 24 h after deviation. Plasma FSH concentrations did not change significantly during the post-ovulatory period. The first significant increase in mean plasma progesterone concentration occurred on the day of follicular deviation. In conclusion, the interval from ovulation to follicular deviation (2.7 days) was similar to that previously reported in B. taurus females, but follicles were smaller. Diameters of the dominant follicle and subordinate follicle did not differ before deviation and deviation was characterized by an increase in dominant follicle and decrease in subordinate follicle growth rate. Variations in FSH concentrations within 12-h intervals were not involved in follicular deviation in Nelore heifers. © 2006 Elsevier B.V. All rights reserved.

The reproductive biology and cycle of Rioraja agassizi (Müller and Henle, 1841) (Chondrichthyes: Rajidae) in southeastern Brazil, SW Atlantic Ocean

Specimens of Rioraja agassizi were collected at Guarujá pier, São Paulo State, Brazil, from March 2005 to March 2006. A total of 275 males were captured. Based on the analysis of the clasper length, gonad weight, clasper gland length, alar thorns and lobule diameter, the size-at-maturity was estimated to be 32 cm. There were 1049 females in the record. Size-at-maturity was calculated in 40 cm. Sexual resting females were observed. Gonadosomatic and hepatosomatic indexes varied significantly in both sexes throughout the year. However, ovulation, egg-laying and presence of sperm in the seminal vesicle were observed all year round. For this reason, an annual cycle with at least one peak in the sexual activity is proposed for this species.

Avaliação da liberação de lh e da concentração de progesterona em protocolos ovsynch e heatsynch em bubalinos (bubalus bubalis)

The objective of this study was to evaluate the LH surge after last hormonal injection of synchronization of ovulation protocols in buffalo. Fifteen multiparous buffaloes received 25 mg of Lecirelin in Day 0, and 150 mg of D-Cloprostenol on Day 7. On Day 8, estradiol benzoate was injected in Group 1 (0.5 mg, n = 5) and Group 2 (1.0 mg, n = 5). On Day 9, five buffaloes received 25 mg of Lecirelin (Control). Blood samples were collected for measure the LH concentrations on Day 7 and then every 3 hours until 72 hours after the PGF 2a injection. For evaluation of LH surge were compared the interval between PGF 2a injection to LH surge, duration, amplitude and area under the LH peak. The LH surge occurred 51.0 + 0.0 hours, 47.3 + 2.7 hours and 47.0 + 3.8 hours after PGF 2a injection for Control, Group 1 and Group 2, respectively (P > 0.05). The duration of LH peak in Control (7.8 + 1.5 hours) was shorter than Groups 1 and 2 (10.5 + 1.5 hours vs. 10.8 + 2.4 hours, respectively; P < 0.05). The amplitudes of LH peak were 4.5 + 0.4 ng/mL, 4.0 + 0.4 ng/mL and 4.3 + 0.8 ng/mL for Control, Group 1 and Group 2, respectively (P > 0.05). The area under LH peak for Control (4.8 ± 0.7) was smaller than the areas of the Groups 1 and 2 (8.8 ± 2.5 vs. 8.7 2.2, respectively; P < 0.05). In summary, the estradiol benzoate injection provided higher duration and area of LH peak than GnRH injection in Ovsynch protocol in buffalo.

Perfil de FSH e LH na divergência folicular em novilhas Nelore (Bos indicus)

Present study aimed to evaluate gonadotropins profiles in 12 Nelore heifers, in order to test the hypothesis that FSH concentrations decrease and LH presents a transient increase during follicle selection. Blood samples from jugular vein were harvested twice daily starting at the time of ovulation (D0) until D5. Plasma samples were assayed for FSH and LH by double antibody radioimmunoassay method. LH and FSH assay sensitivity was 0,02ng/ml and 0,005ng/ml, respectively. The intraassay coefficient of variation was 13,6% and 18,8%, respectively. Data (mean±SEM) were normalized to follicle deviation and analyzed by ANOVA and by linear, cubic, and quadratic regressions. Comparisons between higher and lower FSH values were also performed by T-test. There was no effect of time in plasmatic FSH and LH circulating levels when variance analysis or regression analysis were performed. However, by T-test, FSH concentrationsreached the lowest plasmatic levels 36 (0,40±0,05ng/ml) and 60 hours (0,42±0,04ng/ml) after follicular deviation, comparatively to 36 hours before deviation, when the concentrations were maximal (0,63±0,08ng/ml). In conclusion, there is a FSH decrease, although a transient LH elevation has not been confirmed encompassing follicle deviation in Nelore females.

LH surge in Nelore cows (Bos indicus), after induced estrus or after ovarian superestimulation

Considering that there is limited information about the preovulatory LH surge in Zebu cattle (Bos indicus), the purpose of the present work was to assess the LH surge in Nelore cows during the estrous cycle and after ovarian superestimulation of ovarian follicular development with FSH. This information is particularly important to improve superovulatory protocols associated with fixed-time artificial insemination. Nelore cows (n = 12) had their estrus synchronized with an intravaginal device containing progesterone (CIDR-B ®) associated with estradiol benzoate administration (EB, 2.5 mg, i.m., Day 0). Eight days later all animals were treated with PGF2α (Day 8) in the morning (8:00 h) and at night, when CIDR devices were removed (20:00 h). Starting 38 h after the first PGF2α injection, blood sampling and ovarian ultrasonography took place every 4 h, during 37 consecutive hours. Frequent handling may have resulted in a stress-induced suppression of LH secretion resulting in only 3 of 12 cows having ovulations at 46.7 ± 4.9 and 72.3 ± 3.8 h, respectively, after removal of CIDR-B. Thirty days later, the same animals received the described hormonal treatment associated with FSH (Folltropin ®, total dose = 200 mg) administered twice a day, during 4 consecutive days, starting on Day 5. Thirty-six hours after the first injection of PGF2α, to minimize stress, only seven blood samples were collected at 4 h interval each, and ultrasonography was performed every 12 h until ovulation. In 11 of 12 cows (92%) the LH surge and ovulation were observed 34.6 ± 1.6 and 59.5 ± 1.9 h, respectively, after removal of progesterone source. The maximum values for LH in those animals were 19.0 ± 2.6 ng/ml (mean ± S.E.M.). It is concluded that, in Nelore cows submitted to a ovarian superstimulation protocol, the LH surge occurs approximately 35 h after removal of intravaginal device containing progesterone, and approximately 12 h before the LH surge observed after an induced estrus without ovarian superstimulation. © 2008 Elsevier B.V. All rights reserved.

Avaliaçãeo ultrassonográfica da dinâmica folicular ovariana de jumenta (equus asinus) durante o ciclo estral

The ovarian activity was accessed by ultrasound throughout 10 estrous cycles from Jennies (Marchador Brasileira). Daily ultrasound examinations were performed and the dada was retrospectively studied based on daily identification of each follicle detected. Blood samples were collected every 24 hours from ovulation (D0) until the next identified ovulation. The follicles measuring 11mm were detected and their mean diameter was registered daily using an ovarian map and permitting a retrospective evaluation of the dada, which represented the follicular growth dynamics. One and two major follicular waves were detected in six (60%) and four (40%) cycles respectively from ten estrous cycles in the present study. The primary wave emergency and follicle deviation from the cycles observed during the present study with one major follicular wave occurred at day 10.2 ± 0.75 and at day14.1 ± 0.81 and for cycles with two major waves, those events occurred at 9.0 ± 2.0 and 14.0 ± 1.15 days after ovulation. The maximum diameter of the dominant follicle at the cycles with one and two follicular waves were 37.2 ± 3.35 mm and 37.3 ± 1.1, respectively. The mean intervals from two ovulations were 23.0 ± 1.79 and 22.3 ± 1.26 days when observed in cycles with one and two major follicular waves. The maximum diameter of the dominant follicle was slightly smaller when double ovulations were observed if compared with the single ovulations (P > 0.05). It can be concluded that wave emergency period is observed around 9.6 days of the estrous cycle and the occurrence of deviation can be observed on day 14 of the estrous cycle.

Sincronização do estro e da ovulação utilizando protocolos de curta duração durante a pré- estação reprodutiva em ovelhas Suffolk

This study was carried out with the objective of examining the effect of the short-term estrus synchronization protocol. Ewes were divided in four groups: Control Group (MAP sponges for 12 days, and eCG at withdrawal); Groups I, II and III used the sponge for four days, and 100 μg of PGF was applied at withdrawal; and additionally, Group I (0.1 mg of Estradiol benzoate - EB, in the sponge placement, and in the withdrawn 400 UI of eCG and 50 μg of GnRH 48h later); Group II (35 mg of injectable progesterone and 0.1 mg of EB in the sponge placement, and 400 UI of eCG at withdrawal, and 50 μg of GnRH 48h after); Group III (35 mg of injectable progesterone and 0.2 mg of EB in the sponge placement, and 400 UI of eCG at withdrawal, and 50 ?g of GnRH 56h after). Exams were accomplished for ultrasound and determine the plasmatic concentrations of progesterone and observations of the beginning the estrus and the ovulation. The lack of eCG in Group I caused this protocol to be less efficacious in induction and synchronization of estrus and ovulation. The Control Group had a greater synchronization of estrus and ovulation.

Fatores preditivos do sucesso da inseminação intrauterina: A morfologia espermática de acordo com o novo manual de laboratório da Organização Mundial de Saúde

Objective: The aim of our study was to assess the likelihood of IUI success as a function of the previously described predictive factors, including sperm morphology according to the new reference values defined by WHO. Material and Methods: This retrospective study enrolled 300 couples which underwent IUI. Regression analyses were used to correlate maternal age, number of preovulatory follicles on the day of hCG administration, number of inseminated motile sperm, and normal sperm morphology with clinical pregnancy. Results are expressed as odds ratio (OR) with 95% of confidence intervals (CI). Results: Women older than 35 years showed a lower pregnancy rate (6.5% vs 18.2%, p=0.017). Logistic regression models confirmed the lower chance of pregnancy occurrence for older women (OR: 0.39; CI: 0.16-0.96; p=0.040). The presence of two or more preovulatory follicles on the day of hCG administration resulted in higher pregnancy rate when compared to cases in which only one preovulatory follicle was present (18.6% vs 8.2%, p=0.011). The regression model showed a more than two fold increase on probability of pregnancy when two or more preovulatory follicles were detected (OR: 2.58; CI: 1.22-5.46, p=0.013). The number of inseminated motile sperm positively influenced pregnancy occurrence (OR: 1.47; CI: 0.88-3.14, p=0.027). Similar pregnancy rates were observed when semen samples were classified as having normal or abnormal morphology (10.6% vs 10.2%, p=0.936). Conclusion: Our results demonstrate that sperm morphological normalcy, according to the new reference value, has no predictive value on IUI outcomes. © Todos os direitos reservados a SBRA - Sociedade Brasileira de Reprodução Assistida.

Use of LH or eCG in the last day of superestimulatory treatment

In the last decades several hormonal treatments to induce multiple ovulation and embryo transfer (MOET) have been developed. Tight control of the time of ovulation allowed the use of fixed-time artificial insemination (FTAI) in embryos donors, facilitating animal management. Although, protocols that allow FTAI have evolved and yield as much embryo as conventional protocols that requires estrus detection, substantial increase in viable embryo production has not been observed in superestimulated bovine cattle. The present mini-review put emphasis on superstimulatory protocols in which the last two doses of pFSH are replaced by eCG or LH. Recent results indicate that an extra LH stimulus (using eCG or LH), on the last day of P-36 superestimulatory treatment, seems to improve transferable embryo yield in both Bos taurus and Bos indicus cattle.

Reproductive technology in domestic carnivorous

Background: The delay in development of artificial reproduction techniques on carnivorous could be due to countless reasons, but the lack of commercial interest is probably the most important one. The majority of canines are small structures, canidae are extremely fertile and a great number of species are adapted to domestication or captivity. Finally, the canine gamete physiology presents a difficult adaptation of technology knowledge obtained from other species. Furthermore, domestic felines are animals of company and there is no interest in reproducing them in a large scale, as it has been observed in other domestic animals, however, besides of being a valuable model for the development of in vitro techniques, the domestic cat is also used as an embryo receptor for different species of small wild felines due to physiological similarities among them, in vitro embrionary development, Review: It was reviewed the main insights about the reproductive physiology in female dogs, in vitro oocytary maturation (IVM), pregnancy and conception rate with dogs' frozen/unfrozen semen and PIV in domestic cats. The majority of mammal oocytes restart meiosis spontaneously after ovulation and reaches MII in artificial environment; in an in vitro maturation system in bovines, around 90% of oocytes complete their maturation, although its development capacity can be reduced subsequently. The success of IVM in canidae have been limited, with maturation rate varying from 0 to 58%, usually around 20%. The greatest difficulties include oocyte quality, hormonal environment, protein supplementation, cumulus / oocyte cell interaction, donor breed and age, culture systems, oxygen tension, amino acids, growth factor and sequential means. The freezing process reduces the quality of the semen, firstly because it reduces the number of living sperms and secondly because freezing produces cell modifications that could alter the sperm motility, longevity, integrity of membranes and its fertilizing capacity. Conclusion: Nowadays, several researches are being performed with the aim of increasing viability after dogs' and cats' semen is unfrozen, using extenders, cryoprotectors, freezing and unfreezing curves, addition of antioxidant substances. The aim of this text is to inform about the improvements obtained on the artificial reproduction techniques, emphasizing the oocytary maturation in female dogs, semen cryopreservation and artificial insemination in domestic dogs and cats.

Locus no cromossomo 19 influencia positivamente o número de oócitos recuperados em ciclos estimulados

Objective: To evaluate if identified loci associated with normal age of menopause variation and early menopause can account for the poor response to controlled ovarian stimulation. Methods: A total of 71 patients, with age ≤ 35 years old, undergoing intracytoplasmic sperm injection were tested for three of the four newly identified genetic variants associated with normal variation in menopausal age and early menopause. Patients were divided into two groups: poor responder group (PR group, n=21) and normoresponder group (NR group, n=50). The influence of risk allele frequency on the response to controlled ovarian stimulation (COS) was evaluated. Logistic regression models were used. Results: There was no significant difference in the incidence of the genetic variants between NR and PR group. The risk allele for chromosome 19 variant (rs4806660) demonstrated a protective effect. The presence of a risk allele, either in homozygosis or in heterozygosis, was associated with an increased response to COS, resulting in an elevated number of follicles (Coef: 2.54, P= 0.041) and retrieved oocytes (Coef: 1.41, P= 0.041). Conclusions: Genetic variants rs244715, rs9379896 and rs4806660 are not risk factors for poor ovarian response. Instead, rs4806660 is associated with higher number of follicles and retrieved oocytes. It could be hypothesized that rs4806660 is associated with an increased response to gonadotrophin stimulus. © Todos os direitos reservados a SBRA - Sociedade Brasileira de Reprodução Assistida.