Qualidade e Quantidade de Oócitos Caprinos e Ovinos Recuperados por Aspiração e Fatiamento / Quality and Quantity of Goats and Sheep Oocytes Recovered by Bspiration and Slicing

This study aimed to compare the aspiration techniques and slicing ovaries in order to obtain oocytes, byassessing the quantity and quality of oocytes recovered from goat and sheep females slaughtered in São Luís -MA. The ovaries were collected after slaughtering and kept in saline solution at 0.9%, containing 30 µg / mL ofgentamicin sulfate. In the laboratory the ovaries were randomly divided into two groups with subsequent processof aspiration and slicing. The average rate of oocyte retrieval by aspiration technique were 4,53 oocytes/ovaryin sheeps and 4,11 oocytes/ovary in goats, and by the slicing technique 5,26 oocytes/ovary in sheeps and 5,06oocytes/ovary in goats. It was not observed a significant difference between the recovery rate between species,however there was statistical difference between the techniques on goats, therefore demonstrating that the twotechniques can be used for oocyte retrieval.(AU)

Qualidade e Quantidade de Oócitos Caprinos e Ovinos Recuperados por Aspiração e Fatiamento / Quality and Quantity of Goats and Sheep Oocytes Recovered by Bspiration and Slicing

This study aimed to compare the aspiration techniques and slicing ovaries in order to obtain oocytes, byassessing the quantity and quality of oocytes recovered from goat and sheep females slaughtered in São Luís -MA. The ovaries were collected after slaughtering and kept in saline solution at 0.9%, containing 30 µg / mL ofgentamicin sulfate. In the laboratory the ovaries were randomly divided into two groups with subsequent processof aspiration and slicing. The average rate of oocyte retrieval by aspiration technique were 4,53 oocytes/ovaryin sheeps and 4,11 oocytes/ovary in goats, and by the slicing technique 5,26 oocytes/ovary in sheeps and 5,06oocytes/ovary in goats. It was not observed a significant difference between the recovery rate between species,however there was statistical difference between the techniques on goats, therefore demonstrating that the twotechniques can be used for oocyte retrieval.

Use of the male effect associated with different durations of mating season in goats in the semiarid of Pernambuco / Uso do efeito macho associado a diferentes durações de estação de monta em caprinos no semiárido pernambucano

Background: The goat rearing is practiced mainly in the Northeast region with semiarid climate, since climate conditionsand vegetation are extremely favorable to the species, but it has low productivity due to the low level of technology employed. Management alternatives aimed at maximizing productivity at low cost are essential tools for the reversal of thisframework. Aiming to focus the onset of estrus for females at low cost is common to use the male effect with the breedingseason, representing a practical, inexpensive and efficient methodology for the producer Thus, this study is to evaluate theeffect of different durations of mating season associated with the male effect on reproductive performance of multiparousAlpine goats during the dry and rainy seasons.Materials, Methods & Results: The survey was conducted in the municipality of Serra Talhada-PE from October 2011 toMay 2013. A total of 160 multiparous females aged between 22 and 74 months and four breeders aged between 24 and 38months were used. In the first step, we tested the mating season durations 25 (MS-25) and 45 days (MS-45). In the secondstep, the mating seasons were 25 (MS-25) and 15 days (MS-15) in different climate periods, each experimental groupconsisting of 20 females and one breeder that has been away from females for 15 days before starting the season in orderto stimulate the male effect and thus synchronizing the estrus. The lots were observed at 6:00 and 16:00 pm by qualifiedpersonnel to identify the mated females and after 45 days of the last mating, an ultrasound examination was performed todetect the matrices that had positive pregnancy, assessing the data by the Chi-square test at 5% significance. The estrusresults obtained during the first stage of...(AU)

Use of the male effect associated with different durations of mating season in goats in the semiarid of Pernambuco / Uso do efeito macho associado a diferentes durações de estação de monta em caprinos no semiárido pernambucano

Background: The goat rearing is practiced mainly in the Northeast region with semiarid climate, since climate conditionsand vegetation are extremely favorable to the species, but it has low productivity due to the low level of technology employed. Management alternatives aimed at maximizing productivity at low cost are essential tools for the reversal of thisframework. Aiming to focus the onset of estrus for females at low cost is common to use the male effect with the breedingseason, representing a practical, inexpensive and efficient methodology for the producer Thus, this study is to evaluate theeffect of different durations of mating season associated with the male effect on reproductive performance of multiparousAlpine goats during the dry and rainy seasons.Materials, Methods & Results: The survey was conducted in the municipality of Serra Talhada-PE from October 2011 toMay 2013. A total of 160 multiparous females aged between 22 and 74 months and four breeders aged between 24 and 38months were used. In the first step, we tested the mating season durations 25 (MS-25) and 45 days (MS-45). In the secondstep, the mating seasons were 25 (MS-25) and 15 days (MS-15) in different climate periods, each experimental groupconsisting of 20 females and one breeder that has been away from females for 15 days before starting the season in orderto stimulate the male effect and thus synchronizing the estrus. The lots were observed at 6:00 and 16:00 pm by qualifiedpersonnel to identify the mated females and after 45 days of the last mating, an ultrasound examination was performed todetect the matrices that had positive pregnancy, assessing the data by the Chi-square test at 5% significance. The estrusresults obtained during the first stage of...

Avaliação de oócitos caprinos submetidos à estresse calórico induzido durante a maturação in vitro / Evaluation of Goat Oocytes Subjected to Heat Stress Induced During Maturation In Vitro

Background: The low productivity of Northeast goat herds has been circumvented by the importation of exotic species; however, caution is needed due to the susceptibility of these breeds to the high temperatures found in this region. It is now known that the oocyte and the embryo are the primary targets of the deleterious effects induced by heat stress, causing cellular damage that triggers the cascade of apoptosis. Therefore, the purpose of this study was to evaluate the effect of thermal heat stress during in vitro maturation of oocytes and its effects on embryo production in goats. Materials, Methods & Results: The ovaries were collected in a slaughterhouse and transported to the Laboratory of Biotechnical Reproduction of UFRPE. The cumulus oophorus complexes (COCs) were collected by the technique of slicing the follicles from 2 to 6 mm in diameter, selected based on morphology and placed in a basic medium for maturation. In 10 replications, the COCs were submitted to the thermal heat stress at 41°C for 0 (thermoneutrality at 39°C), 3, 6, 12, 18, and 24 h of maturation in vitro. The data was evaluated in maturation, fertilization, cleavage (D-3), stage of 8-16 cells (D4), morula (D-5), and blastocyst (D-8) after fertilization and blastocysts positive for apoptosis through the TUNEL test. For statistics, the results were expressed as mean and standard deviation. Also considering the measurements addressed in percentages, a comparison of variances was carried out, F-test for variances to the level of signifi cance 5% (P < 0.05). Then, a t-test to compare averages was conducted, to the significance level of 5%, for equivalent variances or distinct variances, according to what was observed in the F-test for variances. A signifi cant difference (P < 0.05) was observed during all time periods studied for heat stress on maturation, fertilization, D-3, D-4, and D-5. On D-8 no significant difference (P > 0.05) was observed between the periods of 3 vs 6 and 18 vs 24 h, and in the blastocysts positive by the TUNEL test for the periods of 0 vs 3, 3 vs 6, 12 vs 18, and 18 vs 24 h of heat stress. Discussion: When applying a thermal shock that produces damage to the oocyte maturation in vitro, the characteristic membrane, chromatin configuration, and meiotic spindles are changed, and thus, the developmental potential of oocytes after fertilization is compromised. It was observed in this study that there was a gradual reduction in the number of oocytes as the time of exposure to heat shock increased, reflecting directly on each stage of IVP embryos. These stages are most vulnerable during maturation in vivo (ovulation), fertilization, within two days after fertilization, and in the first division of cleavage, as evidenced in this study in vitro after heat stress, reducing the number of blastocysts. This suggests that apoptosis can be induced in pre-implantation of embryos exposed to maternal hyperthermia. Moreover, the degree of apoptosis experienced by IVP embryos generally reflects the severity of thermal shock. In this study, the percentage of cells that were TUNEL positive increased with the prolongation of thermal shock. Induction of apoptosis was time dependent and the number of apoptotic cells increased proportionally after 6, 12, 18, and 24 h of exposure. Under the conditions observed in this study, the results indicate that the time in which the oocyte is exposed to heat stress during maturation in vitro is of great importance for embryonic development and their level of apoptotic cells.

Avaliação de embriões ovinos provenientes de oócitos submetidos a estresse calórico durante a maturação in vitro / Evaluation of sheep oocytes submitted to heat stress induced during in vitro maturation

Neste trabalho foi avaliado o efeito do estresse calórico durante a maturação de oócitos sobre a produção in vitro de embriões ovinos. Os ovários foram obtidos em abatedouro e os oócitos colhidos de folículos de 2 a 6 mm de diâmetro. Após seleção, os oócitos, em 10 replicações, foram colocados para maturação in vitro (MIV) durante 24 horas. Os oócitos submetidos ao estresse térmico de 41º C durante 3, 6, 12, 18 e 24 horas foram posteriormente transferidos para completar a MIV a 39 ºC, mesma temperatura utilizada para maturação dos oócitos do grupo controle. O desenvolvimento dos embriões foi determinado nos dias 3, 4, 5 e 8 pós-fecundação. A avaliação da qualidade dos embriões foi efetuada através da contagem total de células coradas pelo DAPI e da determinação do número de blastômeros positivos para apoptose através do teste de TUNEL. Observou-se que o estresse térmico diminuiu (P 0,05) somente até 18 horas de incubação. Os resultados permitem concluir que o estresse calórico durante a maturação in vitro de oócitos reduz a quantidade e a qualidade dos embriões ovinos produzidos in vitro determinadas pela alta incidência de apoptose.

The aim of this work was evaluate the effect of heat stress during oocyte maturation on ovine in vitro embryo production. Ovaries were collected at abattoirs and oocytes retrieved from follicles ranging from 2 and 6 mm in diameter. After selection, all oocytes, in 10 replicates, were placed in in vitro maturation (IVM) during 24 hours. The oocytes were submitted to heat stress of 41º C during 3, 6, 12, 18 and 24 hours and were further transferred to 39º C in order to complete IVM, which was the temperature of maturation of control oocytes. Embryonic development was determined on days 3, 4, 5, and 8 post-fertilization. Embryo evaluation was performed as total cell count by DAPI staining and determination of positive blastomere for apoptosis by the TUNEL assay. We observed that heat stress diminishes (P 0.05) only up to 18 hours of incubation. The results allow the conclusion that heat stress during oocyte in vitro maturation reduces the quantity and quality of ovine embryos produced in vitro determined by the high incidence of apoptosis.

Avaliação de embriões ovinos provenientes de oócitos submetidos a estresse calórico durante a maturação in vitro / Evaluation of sheep oocytes submitted to heat stress induced during in vitro maturation

Neste trabalho foi avaliado o efeito do estresse calórico durante a maturação de oócitos sobre a produção in vitro de embriões ovinos. Os ovários foram obtidos em abatedouro e os oócitos colhidos de folículos de 2 a 6 mm de diâmetro. Após seleção, os oócitos, em 10 replicações, foram colocados para maturação in vitro (MIV) durante 24 horas. Os oócitos submetidos ao estresse térmico de 41º C durante 3, 6, 12, 18 e 24 horas foram posteriormente transferidos para completar a MIV a 39 ºC, mesma temperatura utilizada para maturação dos oócitos do grupo controle. O desenvolvimento dos embriões foi determinado nos dias 3, 4, 5 e 8 pós-fecundação. A avaliação da qualidade dos embriões foi efetuada através da contagem total de células coradas pelo DAPI e da determinação do número de blastômeros positivos para apoptose através do teste de TUNEL. Observou-se que o estresse térmico diminuiu (P < 0,05) a capacidade de maturação dos oócitos de acordo com o tempo de exposição à temperatura de 41º C. No grupo de oócitos incubados a 39 C, 70,70% maturou, enquanto que nos grupos expostos ao estresse térmico, apenas 45,28%, 35,17%, 12,30%, 9,74% e 4,60% maturaram, respectivamente, após 3, 6, 12, 18 e 24 horas de incubação. A duração de exposição dos oócitos ao estresse calórico é inversamente proporcional (P < 0,05) à capacidade de desenvolvimento embrionário e diretamente proporcional (P < 0,05) ao número de blastocistos positivos para apoptose. Todavia, o efeito deletério do estresse térmico sobre a clivagem e os embriões nos estádios de 8 a 16 células e de mórula foi crescente (P > 0,05) somente até 18 horas de incubação. Os resultados permitem concluir que o estresse calórico durante a maturação in vitro de oócitos reduz a quantidade e a qualidade dos embriões ovinos produzidos in vitro determinadas pela alta incidência de apoptose.(AU)

The aim of this work was evaluate the effect of heat stress during oocyte maturation on ovine in vitro embryo production. Ovaries were collected at abattoirs and oocytes retrieved from follicles ranging from 2 and 6 mm in diameter. After selection, all oocytes, in 10 replicates, were placed in in vitro maturation (IVM) during 24 hours. The oocytes were submitted to heat stress of 41º C during 3, 6, 12, 18 and 24 hours and were further transferred to 39º C in order to complete IVM, which was the temperature of maturation of control oocytes. Embryonic development was determined on days 3, 4, 5, and 8 post-fertilization. Embryo evaluation was performed as total cell count by DAPI staining and determination of positive blastomere for apoptosis by the TUNEL assay. We observed that heat stress diminishes (P < 0.05) oocyte maturation capacity in accordance with exposure time of 41º C. In the group of oocytes incubated at 39 ºC, 70.70% matured, while in the groups exposed to heat stress of 41º C, only 45.28%, 35.17%, 12.30%, 9.74% and 4.60% matured, respectively, after 3, 6, 12, 18 and 24 hours of incubation. The duration of exposure to heat stress is inversely proportional (P < 0.05) to embryonic developmental capacity directly proportional (P < 0.05) to the number of blastocysts positive to apoptosis. However, the cleavage rate and embryonic development from 8 to 16 cells and morulae stages were affected by heat stress (P > 0.05) only up to 18 hours of incubation. The results allow the conclusion that heat stress during oocyte in vitro maturation reduces the quantity and quality of ovine embryos produced in vitro determined by the high incidence of apoptosis.(AU)