M. A. L. Oliveira

Effects of retinol on the in vitro development of Bos indicus embryos to blastocysts in two different culture systems.

The objective of this study was to evaluate the effect of retinol (RT) and retinoic acid (RA) on the in vitro development of pre-implantation goat embryos cultured in potassium simplex optimized medium or synthetic oviduct fluid or cocultured in oviductal cells monolayer either in potassium simplex optimized medium or synthetic oviduct fluid. A total of 2407 cumulus-oocyte complexes were aspirated from 2 to 6 mm ovarian follicles from slaughtered animals. Selected cumulus-oocyte complexes were subjected to in vitro maturation in TCM 199 for 24 h at 39 °C in an atmosphere of 5% (v/v) CO(2) in humidified air. In vitro fertilization was performed in modified defined medium. Eighteen hours after in vitro fertilization, cumulus cells were removed and presumptive zygotes were randomly distributed into experimental groups. In Experiment 1, presumptive zygotes were cultured in potassium simplex optimized medium, potassium simplex optimized medium + RT, potassium simplex optimized medium + retinoic acid, synthetic oviduct fluid, synthetic oviduct fluid + RT and synthetic oviduct fluid + RA at 39 °C in a humidified atmosphere of 5% (v/v) CO(2), 5% (v/v) O(2) and 90% (v/v) N(2). In Experiment 2, presumptive zygotes were cocultured in potassium simplex optimized medium + oviductal cells monolayer, potassium simplex optimized medium + RT + oviductal cells monolayer, potassium simplex optimized medium + RA + oviductal cells monolayer, synthetic oviduct fluid + oviductal cells monolayer, synthetic oviduct fluid + RT + oviductal cells monolayer and synthetic oviduct fluid + RA + oviductal cells monolayer in an atmosphere of 5% (v/v) CO(2) in humidified air. In both experiments, media were partially changed on day 2 after in vitro fertilization and unfertilized oocytes were excluded from the experiment. Embryos were cultured or cocultured for 8 days. In Experiment 1, there was no effect of RT or RA supplementation on the proportion of oocytes that reached the morula or blastocyst stages. By contrast, Experiment 2 demonstrated that the addition of 0.28 μg/ml RT and 0.5 μm RA to the embryo culture media stimulated (p < 0.05) development to the morula and blastocyst stages under the coculture conditions tested. In conclusion, retinoids play an important role in pre-implantation development of goat embryos and can be used to enhance in vitro embryo production.

Sexing of Dorper sheep fetuses derived from natural mating and embryo transfer by ultrasonography

In order to improve fetal sexing in the Dorper sheep breed, the objective of the present study was to determine, by repeated ultrasonographic examinations, the migration period of the genital tubercle (GT) in sheep fetuses derived from natural mating or embryo transfer and to compare the accuracy of a single examination with repeated examinations at short intervals. For this purpose, transrectal ultrasound was performed, using a double-frequency linear transducer (6.0 and 8.0 MHz) for monitoring 51 sheep fetuses distributed in three experimental groups (EI, EII and EIII). The fetuses in EI (n = 23) and EII (n = 18) derived, respectively, from natural mating and embryo transfer were monitored at 48-h intervals from the 30th to 60th day of gestation and sexed based on the final location of the GT. The fetuses in EIII (n = 10), which originated from embryo transfer, were examined only once on the 65th day of gestation and sexed taking into consideration the final position of the GT and/or by identification of anatomical structures of external genitalia. The accuracy of fetal sexing was 91.3% (21 fetuses sexed/23 quantified) in EI, 88.9% (16 sexed/18 quantified) in EII and 100% (10 sexed/10 quantified) in EIII, without significant difference (P > 0.05) between experiments. Migration of the GT occurred earlier (P < 0.05) in fetuses produced by natural mating (43.0 +/- 2.8 days) than in those derived from embryo transfer (46.1 +/- 4.7 days). The results show that fetal sexing can be done from the 50th day onward in fetuses produced by natural mating and from the 60th day onward in fetuses derived from frozen embryos. It can also be concluded that repeated ultrasonographic exams in short time intervals do not maximise the accuracy of fetal sexing. In addition, real-time ultrasonography is a reliable tool for fetal sex determination in sheep after Day 50 of gestation, taking into account both the location of the GT and the identification of external genital structures.

The use of steroid hormones in superovulation of Nelore donors at different stages of estrous cycle.

The objective of the present study was to evaluate the superovulatory response and ova/embryo recovery from Nelore donors following treatment with a controlled internal drug releasing device and estradiol benzoate (CIDR-B program) at different stages of the estrous cycle. The control group (TI; n=40) received a standard superovulation protocol with females of this group being between days 9 and 12 of the estrous cycle (estrus = day 0). The donors that received a CIDR-B program containing 1.9 g progesterone and an intramuscular injection of estradiol benzoate (2 mg) were at day 0 (TII; n=30), between days 2 and 6 (TIII; n=30), days 7 and 12 (TIV; n=30), days 13 and 16 (TV; n=30) and days 17 and 20 (TVI; n=30) of the estrous cycle. Superovulation was induced with 400 IU of p-FSH, divided into eight decreasing doses (80/80; 60/60; 40/40; 20/20) at intervals of 12h. The donors received PGF2alpha (Cloprostenol) 48 h after beginning the treatment and CIDRs were removed 12h later. Artificial inseminations (AI) were performed 12 and 22 h after the initiation of estrus and embryos were collected 7 days after AI. The mean numbers (+/-S.E.M.) of total ova and embryos, viable (transferable) and degenerated embryos were 14.2+/-11.3, 7.4+/-6.9 and 3.2+/-3.5 (TI), 13.3+/-10.4, 7.1+/-6.2 and 3.3+/-4.3 (TII), 13.5+/-7.0, 8.1+/-6.7 and 2.3+/-3.0 (TIII), 17.4+/-9.9, 9.4+/-6.9 and 4.0+/-4.4 (TIV), 16.9+/-8.8, 9.8+/-8.1 and 2.7+/-2.5 (TV) and 13.0+/-7.8, 7.2+/-6.9 and 2.3+/-2.5 (TVI), with no significant differences (P>/=0.05) among groups. Pregnancy rates of 67.1% (TI; n=86/128), 60.8% (TII; n=59/97), 62.5% (TIII; n=73/115), 64.1% (TIV; n=84/131), 72.3% (TV; n=81/112) and 60.6% (TVI; n=63/104) were obtained with embryos transferred from these collections and did not differ significantly (P>/=0.05) among groups. The results of the present study allow us to conclude that a combination of steroid hormones may be used prior to superovulation in Nelore donors, at any stage of the estrous cycle without affecting the efficiency of embryo transfer programs.

Reliability of ultrasound for early sexing of goat fetuses derived from natural mating and from fresh, frozen and vitrified embryo transfer.

The aim of the present study was to identify the migration period of the genital tubercle and its later differentiation into external genital structures in fetuses derived from natural mating and fetuses from fresh, frozen and vitrified embryo transfer. A transrectal ultrasound with a double-frequency linear transducer (6.0 and 8.0 MHz) was used to monitor 123 goat fetuses, which were allocated to one of four groups: fetuses originating from controlled natural mating (G1, n = 32) and fetuses derived from fresh (G2, n = 34), frozen (G3, n = 30) and vitrified (G4, n = 27) embryo transfer. The transferable embryos were collected 7 days after mating by laparoscopy. Migration of the genital tubercle occurred significantly earlier (P < 0.05) in G1 than in G2, G3 and G4. The visualisation of the scrotum, prepuce and vulva occurred significantly earlier (P < 0.05) in G1 than in G2, G3 and G4. Our results show that fetal sexing is feasible after 55 days for fetuses from natural mating and after 60 days in fetuses from fresh and cryopreserved embryos. Thus, real-time ultrasonography is a reliable tool for fetal sex determination in goats after Day 50 of pregnancy, taking into account both the location of the genital tubercle and the identification of external genital structures.

Pulmonary Thromboembolism and Thrombus in Patent Foramen Ovale

Patent Oval Foramen (FOP) is the persistence of the opening between the atrial primum and atrial secundum in the location of the oval fossa present in about 25% of the adult population1. In generally, it is a casual finding and without hemodynamic repercussions. However, the paradoxical embolism as a cause of ischemic events due to the patency of this foramen is documented since the nineteenth century. Recent studies have shown the association between FOP and various medical conditions, some potentially serious, such as ischemic stroke (AVCi) and pulmonary embolism (TEP).