Efficiency of OPU-IVEP-ET of Fresh and Vitrified Embryos in Buffaloes

The present study aims to report ovum pickup (OPU), in vitro embryo production (IVEP) and embryo transfer (ET) outcomes of fresh and vitrified buffalo embryos. For this purpose, 36 buffalo donors were submitted to 11 OPU sessions (n = 201). A total of 998 oocytes (5.0 +/- 0.5/donor/session) and 584 viable oocytes (2.9 +/- 0.3/donor/session) were recovered. Viable oocytes (grades 1, 2 and 3) were subjected to IVM, IVF (D0) and IVC. On D2, 54.5% of cleavage rate was obtained. Embryo yield on D7 was 44.9% (grade 1: 229 embryos, grade 2: 5 embryos and grade 3: 28 embryos). From this total, 115 fresh (grades 1 to 3) and 70 vitrified embryos (only grade 1) were transferred into recipients previously synchronized with fixed time embryo transfer (FTET) protocol. Vitrification was performed using the cryotop method. Pregnancy diagnosis in fresh and in vitrified groups were, respectively: 43.5% (50/115) and 37.1% (26/70) on 30 days after embryo transfer, and 41.7% (48/115) and 31.4% (22/70) on 60 days after embryo transfer. In conclusion, our results demonstrate the possibilities for commercial use of the techniques of OPU, IVEP and ET of fresh and vitrified embryos in buffaloes.

Influence of Parity and Season of the Year on Oocyte Quality and Number in Buffaloes

The aim of the present study was to evaluate the effects of season of the year (summer and winter) and parity (heifers and cows) on oocyte quality and number in buffaloes. For this purpose, 71 buffaloes had follicular wave emergence synchronized before OPU. OPU of all follicles >= 2mm was done 5 days after the beginning of the hormonal protocol, in 4 replicates (two for each season). Data were analyzed by ANOVA using PROC GLIMMIX, in a 2 x 2 factorial arrangement of treatments. No interactions were observed in following variables: number of follicles, number of total and viable oocytes, recovery rate, percentage of viable oocytes, grade I oocytes, grade II oocytes, grade III oocytes, denuded oocytes, expanded cumulus oocytes, and atretic/degenerated oocytes. Number of follicles visualized at OPU and recovery rate were not affected by parity or season. Relative to parity, number of total and viable oocytes were greater in heifers than in cows, respectively. Concerning season of the year, number of viable oocytes and viable oocyte rate were increased in winter. In conclusion, better oocyte quality can be obtained from heifers and during winter in buffaloes. However, the number of total oocytes seems to be more influenced by parity than by season of the year in this species.

Efeitos da redução ou substituição do soro fetal bovino por outros compostos na maturação in vitro de oócitos bovinos

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Dinâmica da vitelogênese durante a maturação ovariana em Artemesia longinaris

Pós-graduação em Aquicultura - FCAV

Population Dynamics and Growth Rates of Endosymbionts During Diaphorina citri (Hemiptera, Liviidae) Ontogeny

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Histopathological study of ovaries of Rhipicephalus sanguineus (Acari: Ixodidae) exposed to different thymol concentrations

Thymol is a monoterpene with proven acaricide action for several tick species. In addition to killing these ectoparasites, thymol can also reduce oviposition and egg hatch rate. However, the effects of thymol on the morphophysiology of tick ovaries are still unknown. Thus, the aim of this study was to evaluate the morphophysiological changes caused by this active principle in ovaries of Rhipicephalus sanguineus after a 6-day feeding period, through the application of morphohistochemical techniques. After the feeding period, a total of 50 females were divided into five groups and immersed in the following solutions: (I) distilled water (control), (II) 30 % ethanol (control), (III) 1.25 mg/mL thymol, (IV) 2.5 mg/mL thymol, and (V) 5.0 mg/mL thymol. The experimental groups were kept in a climatic chamber (27 +/- 1 A degrees C; RH 80 A +/- 10 %) for 5 days. After this period, morphological (hematoxylin/eosin) and histochemical (von Kossa) techniques were applied after remotion of the ovaries. The morphological results revealed large vacuoles in germ cells at different developmental stages and invaginations that represent deformations in the chorionic membrane. From the results obtained in this study, it was concluded that thymol interfered with the development of oocytes, which showed degeneration signs. The treatment containing 5.0 mg/mL thymol affected more accentuately the morphological development. Moreover, thymol also altered the calcium content of yolk granules, which generally showed an intense staining for this element.

Oocyte maturation in the sloth's giant tick Amblyomma varium (Acari: Ixodidae) in an ecological context

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Moments of induced spawning and embryonic development of Brycon amazonicus (Teleostei, Characidae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Fecundity and fertility in a freshwater population of the neotropical amphidromous shrimp Macrobrachium acanthurus from the southeastern Atlantic

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Functional morphology of the reproductive system and sperm transfer in Stenopus hispidus (Crustacea: Decapoda: Stenopodidea), and their relation to the mating system

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Control of oocyte maturation

Oocyte maturation is a complex process involving nuclear and cytoplasmic maturation. The nuclear maturation is a chromosomal segregation and the cytoplasmic maturation involves the reorganization of the cytoplasmic organelles, mRNA transcription and storage of proteins to be used during fertilization and early embryo development. The mechanism of oocyte maturation in vivo and in vitro still are not totally understood. However it is generally accepted that the second messenger cyclic adenosine monophosphate (cAMP) plays a critical role in the maintenance of meiotic blockage of mammalian oocytes. A relative increase in the level of cAMP within the oocyte is essential for maintaining meiosis block, while a decrease in cAMP oocyte concentration allows the resumption of meiosis. The oocyte cAMP concentration is regulated by a balance of two types of enzymes: adenylate cyclase (AC) and phosphodiesterases (PDEs), which are responsible for the synthesis and degradation of cAMP, respectively. After being synthesized by AC in cumulus cells, cAMP are transferred to the oocyte through gap junctions. Thus, specific subtypes PDEs are able to inhibit or attenuate the spontaneous meiotic maturation of oocytes with PDE4 primarily involved in the metabolism of cAMP in granulosa cells and PDE3 in the oocyte. Although the immature oocytes can resume meiosis in vitro, after being removed from antral follicles, cytoplasmic maturation seems to occur asynchronously with nuclear maturation. Therefore, knowledge of the oocyte maturation process is fundamental for the development of methodologies to increase the success of in vitro embryo production and to develop treatments for various forms of infertility. This review will present current knowledge about the maintenance of the oocyte in prophase arrest, and the resumption of meiosis during oocyte maturation, focusing mainly on the changes that take place in the oocyte.

Biotecnologias aplicadas à reprodução de éguas velhas

The acceptance of biotechnology for the most equine breeders association had a significant effect in the horse industry, gaining popularity around the world, because the increasing on the genetic gain, allowing the use of sub fertile mares and stallions with high genetics value on reproduction. The embryos in vitro production of human and cattle has been used with success, however in vitro embryo production is not efficient in the horse, as oocyte transfer (OT) and intracytoplasmatic sperm injection (ICSI). The oocyte transfer has been used especially in subfertile old mares presenting reproductive pathologies as: endometrite, cervical and uterine adhesions, blocked oviduct, perineal laceration and ovulation failures. During oocyte recovery process, the oocytes must be collected from immature follicles that need be matured in vitro or in vivo matured oocytes from pre-ovulatory follicles through the transvaginal aspiration guided by ultrasound. The recovered oocyte is transferred to a previously inseminated recipient mare, through the flank laparotomy. The intracytoplasmatic sperm injection (ICSI) is a procedure of in vitro fertilization that needs only one sperm that is aspirated and injected inside the oocyte. The oocytes used, can be from mature and immature follicles. Fresh, cooled and frozen semen can be used, because the procedure not requires a functional sperm. The use of Piezo drill resulted in a breakthrough the pellucid zone, allowing the vibration per minute provided in the sperm injection pipette, a major result of cleaved oocytes, due to a better sperm injection in the oocyte. The embryo transfer can be straight inside the oviduct, as also transcervical transferred after embryo culture produced in vitro. In conclusion both procedures (OT and ICSI) are effective to be used on equine assisted reproduction, getting results even lower than expected, but satisfactory from animal genetically superior

Influência da adição de fator de crescimento fibroblástico 10 (FGF10), durante a maturação oocitária, na produção in vitro de embriões bovinos

The Brazilian livestock stands out for having the world largest commercial herd of cattle and leads meat exportation and production of bovine embryos. The in vitro production (IVP) of embryos is considered an effective option to overcome problems such as infertility in cows with high economic value and also for genetic improvement of cattle. The in vitro oocyte maturation is an essential step to the success of IVP, but is still considered poor when compared to in vivo maturation. Recent studies have suggeested an important role of Fibroblast Growth Factor 10 (FGF10) on the in vitro maturation of oocytes, which favored the expression of genes related to oocyte maturation and cumulus cell expansion. Aware that maturity stage influences the final production of blastocysts, we aimed study to verify if the addition of FGF10 into the maturation medium is able to affect positively the IVP of bovine embryos. Hence, FGF10 was added to maturation in five different concentrations: 0.5 ng/mL (group 0.5), 2.5 ng/mL (group 2.5), 5 ng/mL (group 5), 10 ng/mL (group 10) and 50 ng/mL (group 50). Additionally, two other maturation groups were used, group BSA (Bovine Serum Albumin, 4 mg/mL) and group FCS (Fetal Calf Serum, 10%). The rates of cleavage, morula and blastocyst were analyzed by Analysis of Variance (ANOVA), differences of P<0.05 were considered significant. Cleavage rates did not differ between the seven groups. On the other hand, morula rate on FCS group was higher than groups BSA, 0.5, 10 and 50 (P<0.05), but did not differ among groups treated with intermediate doses of FGF10 (2.5 and 5). FCS group presented higher blastocyst rate compared to all other groups that were well below the FCS group (P<0.0001). Therefore, the use of FGF10 during oocyte maturation did not affect positively embryo development on the IVP of bovine embryos

Estresse término na reprodução de vacas leiteiras

Climate change in tropical countries, like Brazil, causes major problems in dairy production due to an increase of heat stress effects. In recent years, milk production in Brazil increased 36.07%. The Southeast region remains a leader in production with herds of high producing Holstein cattle (mostly), which is more susceptible to heat stress. Thermal stress decreases fertility in direct and indirect ways. Conception rates are reduced of 40-60% during cooler months of the year and 10-20% in the warmer months. Negative effects of heat stress involve changes in reproductive hormones, follicular development, oocytes, and embryos, and decreased dry matter intake. Several studies discuss change in reproductive hormones, such as reduction in plasma concentration of GnRH, LH, and oestradiol, which lead to decreased detection of estrus and ovulation. Various methods are being studied to bypass these negative effects and increase the fertility of dairy cows under heat stress. Cooling systems are the most advantageous and can be associated with technologies such as ET and TAI

Criopreservação de embriões bovino produzidos in vitro: estado da arte

In the last years, the embryo in vitro production for every domestic species and mainly for bovine has attained a notorius status. This reproductive biotechnical procedure associate with ultrasound-guided ovum pick up (OPU) has been more and more incorporated and spread in our cattle herds, ranking up Brazil already at the top of the list in number of in vitro embryo produced. Some significant advantages provided, such as the possibility of using the premature or pregnant animals oocytes, without necessarily requiring the use of hormonal treatment, to make it possible to generate pregnancy at a shorter period of time, the rationalization in the use of semen and optimization in the use of sexed semen were determinant factors for OPU/IVP to reach this outstanding position. Nevertheless, right now the possibility of IVP embryo cryopreservation, just now is the biggest impediment for maximizing the use of this biotechnology, due to both lack of efficient methods and low laboratory produced embryo cryotolerance. Nowadays, the most used methods of IVP embryo cryopreservation are: slow freezing and vitrification. Traditionally, slow freezing is still the most used methods for in vivo and in vitro produced embryo cryopreservation. However, more recently vitrification - although still not commercially used in large scale - has been presenting satisfactory results in IVP embryo cryopreservation, according to searches