Protocolos hormonais de indução de estro sincronizado em ovelhas leiteiras na contra estação reprodutiva.

[Protocols hormonal to estrus induction and synchronization and pregnancy rate in dairy ewes out-of season mating]

Cervical dilatation in Santa Ines ewes induced with misoprostol, oxytocin and estradiol for development of non-surgical method for embryo recovery.

The objective of this study to develop a pharmacological protocol cervical dilatation in ewes Santa Ines aimed at collecting embryos transcervical. Multiparous ewes were used (n=30), cyclic, aged 24 and 48 months, with an average weight of 50.7 ± 5.4 kg and body scores between 2.75 and 3.5 (1-5) in design experimental model in "crossover". The study proved the possibility of increasing the cervical transposition rate in ewes Santa Ines with the use of hormonal associations, enabling the collection of embryos transcervical in sheep, avoiding surgery.Proceedings of the 29th Annual Meeting of the Brazilian Embryo Technology Society (SBTE); Gramado, RS, Brazil, August 20th to 23rd, 2015, and 31st Meeting of the European Embryo Transfer Association (AETE); Ghent, Belgium, September 11th and 12th, 2015

Comportamento sexual de ovelhas superovuladas submetidas a dois métodos de acasalamento.

[Sexual behavior of superovulated sheep subjected to two methods of breeding]

Use of two doses of cloprostenol at different intervals for estrus synchronization in Santa Inês ewes.

Abstract: The objective of this study was to compare protocols for estrus synchronization, using two doses of cloprostenol in different intervals, in Martch, during the breeding season, in Santa Inês ewes, in the city of Cachoeiras de MacacuRJ. A total of 30 ewes (43.9±6.4 kg, 2.9±0.27 BCS and 3.4±1.6 years old) weaned at least for three months was equally allocated into three treatments, with intervals of: 11.5 days (G11.5: n=10), 9 days (G9: n=10) or 7 days (G7: n=10). The dose used for each administration was 37.5 µg cloprostenol (Estron®, Agener União, São Paulo, Brazil) intramuscularly. Transrectal ultrasound evaluations were performed (B-mode; SonoScape®, Shenzhen, China) for monitoring the follicular and luteal dynamic, daily from 5 d before the first dose, and every 12 h after both administrations, again for 5 d or until ovulation. For estrus detection, females were teased individually every 12 h after each cloprostenol administration for 5 d and for 5 min per animal. Normal quantitative variables were subjected to ANOVA followed by Tukey test (P0.05) among groups: G11.5 60% (6/10); G9 80% (8/10) and G7: 80% (8/10) as well as the duration of estrus (G11.5: 24.0±13.1 h; G9: 37.5±7.7h and G7: 28.5±11.0 h. After the second dose, estrus presentation rates and duration of estrus also did not differ (P>0.05) among groups, respectively: G11.5 90% (9/10) and 29.3±12.2 h; G9 ? 100% (10/10) and 36.0±10.4 h; and G7 80% (8/10) and 31.5±8.9 h. There was no statistical difference (P>0.05) in the intervals from the second dose to the onset of estrus, end of estrus, and from estrus to ovulation between G11.5 (48.5±8.9 h; 80.0±8.5 h; 35.0±20.1 h), G9 (50.3±13.1 h; 83.0±9.1 h; 25.5±12.2 h) and G7 (36.5±6.2 h; 68.0±14.0 h and 20.3±6.1 h). The interval from the administration of the second dose to ovulation differed (P<0.05) among groups G11.5 (78.7±9.4 h), G9 (75.5±8.3) h e G7 (56.8±6.2 h). The G7 anticipated ovulation and presented a lower standard deviation (P<0.05). In conclusion, it appears that animals in the G7 were close to follicular dominance period or were already with the dominant follicles while G11.5 and G9 might still be in the beginning of the follicular wave. The three protocols were effective for estrus synchronization in Santa Inês ewes.Proceedings of the 29th Annual Meeting of the Brazilian EmbryoTechnology Society (SBTE); Gramado, RS, Brazil, August 20th to 23rd, 2015, and 31st Meeting of the European Embryo Transfer Association (AETE); Ghent, Belgium, September 11th and 12th, 2015

The influence of different methods of frozen-thawed ovine spermatozoa selection on sperm capacitation and viability after incubation.

Abstract: Sperm capacitation is an essential event for fertilization; however, it decreases the sperm lifespan and viability. The aim of this study was to evaluate the effects of four sperm selection techniques on sperm capacitation and viability after incubation. A pool of frozen-thawed sperm from 10 Santa Inês rams was used. The samples were submitted to one of the following sperm selection techniques: sperm washing, Percoll gradient, mini-Percoll gradient, Swim-up and control group. At mini-Percoll technique, was used 400 microliters of 90% and 45% gradients and a centrifugation at 500 xg for 5 minutes. In Percoll, was used 1 mL of each gradient and a centrifugation at 700 xg for 10 minutos. During Swim-up, the sperm was incubated in 1 ml of SPERM-TALP for 45 minutos in humidified atmosphere at 37.5oC. Finaly, at sperm washing the sample suffered centrifugation at 300 xg for 8 minutes, using SPERM-TALP. At the end of each treatment, the selected spermatozoa were incubated at 37oC for 1 h, 2 h, and 3 h. Viability was assessed using acridine orange-propidium iodide combination by computer-assisted sperm analysis. Capacitation status was evaluated using chlortetracycline staining and observed under epifluorescence microscopy. Data were analyzed by ANOVA, followed by Tukey test (P0.05) among Percoll (36%), mini-Percoll (34%) and Swim-up (30%), and were lower (P0,05) entre Percoll (36%), mini-Percoll (34%) e Swim-up (30%), as quais foram inferiores (P<0,05) ao grupo controle (47%) e lavagem por centrifugação (41%). A taxa de espermatozoides não capacitados foi superior (P<0,05) no momento 0 h (12%) e diminuiu após 3 h (1,5%), independentemente do método. Houve interação (P<0,05) entre intervalo de incubação e método na taxa de espermatozoides reagidos. O índice de reagidos foi menor (P<0,05) em 0 h (50%) e 1 h (53%) e maior após 3 h (64%). Percoll (60%) e mini-Percoll (61%) apresentaram maiores valores para reagidos (P<0,05) enquanto o controle apresentou o menor (49%). Observou-se interação (P<0,05) entre intervalo de incubação e método na vitalidade espermática. A taxa de íntegros em 0 h foi a maior (17,5%; P<0,05), após os diferentes métodos. O Swim-up apresentou maior taxa de íntegros (17,4%; P<0,05), independentemente do intervalo de incubação. Em conclusão, o intervalo de incubação interfere nos padrões de capacitação e vitalidade de espermatozoides ovinos congelados. A seleção espermática aumenta a taxa de células reagidas e o Swim-up permite maior vitalidade durante a incubação.Proceedings of the 29th Annual Meeting of the Brazilian Embryo Technology Society (SBTE); Gramado, RS, Brazil, August 20 to 23, 2015. Abstracts

GnRH potential to synchronize follicular emergence and ovulation prior to superovulatory day 0 protocol in sheep.

The role of GnRH to synchronize ovulation and follicular emergence previous to superovulatory protocol, started on the first day of the estrous cycle (Day 0), was assessed in Santa Inês ewes. For estrus synchronization, 60 mg medroxyprogesterone acetate sponges were used for 6 d plus 37.5 ?g d-cloprostenol and 300 IU eCG at fifth day. After sponge removal, ewes were assigned to three treatments: GControl ? saline at 12 h (n = 10); G24h ? GnRH at 24 h (n = 10); or G36h ? GnRH at 36 h (n = 9). In conclusion, considering the beneficial effects of G36h in synchronizing ovulation and to promote the absence of dominant follicles in the first days of estrous cycle. According to data obtained, the best time to start the superovulatory treatment, known as "Day 0", could be 80 h after sponge removal (56 h for the occurrence of ovulation plus 24 h to reset dominant follicles), in the induction of synchronized estrus, for Santa Inês ewes.Proceedings of the 29th Annual Meeting of the Brazilian Embryo Technology Society (SBTE); Gramado, RS, Brazil, August 20th to 23rd, 2015, and 31st Meeting of the European Embryo Transfer Association (AETE); Ghent, Belgium, September 11th and 12th, 2015

GnRH use in different times on estrus synchronization and ovulation in Santa Inês ewes.

Proceedings of the 28th Annual Meeting of the Brazilian Embryo Technology Society (SBTE), August, 2014, Natal, RN, Brazil. Abstracts

Use of D-Cloprostenol at different intervals of administration for estrous synchronization in cyclic Saanen goats.

Proceedings of the 28th Annual Meeting of the Brazilian Embryo Technology Society (SBTE), August, 2014, Natal, RN, Brazil. Abstracts

Ultrasound diagnosis and macroscopic post-mortem evaluation of hydrosalpinx in dairy goats: a case report.

The oviduct plays a key role in the reproduction providing an ideal environment for the oocyte maturation, sperm capacitation, fertilization, and gamete and embryo transport. Hydrosalpinx is a pathology that can occur uni or bilaterally in the oviduct, affecting the reproductive function. In goats, it is considered of low frequency. No malformation or segmental aplasia of the organ was detected. Adhesions were verified in both ovaries. It was concluded that the transrectal US exam associated with reproductive history represents an efficient tool in diagnosing hydrosalpinx in dairy goats.Abstracts on VI International Symposium on Animal Biology of Reproduction, November 6-9, 2016, Campos do Jordão, SP, Brazil

Comparison of different sperm selection techniques in ram frozen-thawed sperm.

Resumo: Background: The success of fertilization is directly associated with semen quality and the sperm preparation. Considering the common use of cryopreserved spermatozoa, there is a need to develop strategies for sperm preparation in order to achieve a sperm sample of high quality through a rigorous selection of sperm. Thus, sperm cells are being more extensively investigate. This study aimed evaluating the influence of different sperm selection techniques on ram sperm parameters in semen preparation. Materials, Methods & Results: Frozen-thawed commercial semen from 10 Santa Inês rams was subjected to either: swim-up, Percoll, mini-Percoll, sperm washing by centrifugation or a control group. After each technique, samples were incubated at 37°C for 1 h, 2 h and 3 h. At post-selection moment (0 h) and at each interval, sperm recovery rate, motility, capacitation and plasma membrane (PM) integrity were analyzed. The lowest (P 0.05). Most part of motility parameters were not affected (P > 0.05) by the technique at 0 h; just swim-up obtained higher (P 0.05), regardless of interval incubation. However, the latter three techniques presented more (P 0.05). Discussion: The present study evaluated the sperm cell and its preparation for receiving the oocyte under optimal conditions. evaluate the sperm cell and its preparation for receiving the oocyte under optimal conditions. Although the swim-up technique promoted higher rates for some of the sperm parameters evaluated, Percoll protocols are the most widely used procedures for selection during the sperm preparation in many species, possibly because of its greater speed, practical method and convenience compared to the swim-up technique. Percoll and mini-Percoll recovered approximately 10 times more cells than swim-up, which is an important feature to be considered during sperm preparation for in vitro fertilization (IVF), being possible to use only one semen straw. The high capacitated and acrosome reaction rates observed after the treatments in the current study, are probably reinforced by changes in sperm cells caused by the cryopreservation process. In order to strength the evidence that frozen-thawed sperm, even after selection, is sensible and reactive to capacitation-like events, we demonstrated the capacitated and acrosome-reacted cell values immediately after the selection treatments behaved differently than when authors used ram fresh sperm. Possibly, this capacitation-like changes observed in frozen-thawed sperm occurs regardless of the selection treatment used. Analyzing the motility parameters immediately after the selection, all treatments maintained or increased the rates compared to the control group. The swim-up, mini-Percoll and Percoll did not differ in any parameters. Given that Percoll and mini-Percoll did not show differences in relation to swim-up for motility parameters, such techniques can be used to replace the latter, obtaining similar sperm samples with good quality. However, swim-up technique involves a procedure that recovers a clean fraction without debris and other types of cells, with high rate of mobile sperm with excellent quality, reason why it can justify the higher recovery of intact spermatozoa after the technique