Effect of different gonadotropins on in vitro maturation of sheep oocytes / Efeito de diferentes gonadotrofinas na maturação in vitro de oócitos ovinos

The aim of this study was to examine the effect of replacing the use of follicle-stimulating hormone (FSH) with equine chorionic gonadotropin (eCG) and human chorionic gonadotropin (hCG) on the in vitro maturation (IVM) of sheep oocytes. After sheep ovaries were collected (n=300), the cumulus-oocyte complexes were aspirated, selected, and divided into four groups according to the IVM medium: CON group, in which the basic IVM medium was used; and eCG, hCG, and FSH groups, in which the oocytes were immersed in basic IVM medium with 10 IU/mL eCG, 10 IU/mL hCG, and 10 µg/mL FSH-p, respectively. In vitro maturation of the oocytes was performed at 38.5 °C, in a humidified atmosphere of 5% CO2 in air, for 24 h. Subsequently, the oocytes were evaluated for the degree of cumulus-cell expansion, chromatin configuration, GSH levels, and active mitochondria. There were no significant differences for the rate of cumulus cell expansion. The percentage of oocytes in MII was higher in the eCG group than in the CON and hCG groups (P<0.05) and similar to that of the FSH group. In conclusion, eCG can be used as a substitute for FSH in IVM of sheep oocytes.

O objetivo deste estudo foi avaliar o efeito da gonadotrofina coriônica equina (eCG) e da gonadotrofina coriônica humana (hCG), em substituição ao uso de hormônio folículo estimulante (FSH) na maturação in vitro (MIV) de oócitos ovinos. Após a coleta de ovários (n=300) ovinos, os complexos cúmulus-oócitos (CCOs) foram aspirados, selecionados e divididos em quatro grupos de acordo com o meio de MIV: grupo CON, em que foi utilizado o meio MIV base; e grupos ECG, HCG e FSH, em que os oócitos foram imersos em meio MIV base adicionado de 10 UI/mL de eCG, 10 UI/mL de hCG e 10 µg/mL de FSH-p, respectivamente. A MIV dos oócitos foi realizada a 38,5°C, em atmosfera umidificada de 5% de CO2 em ar, durante 24 horas. Posteriormente, os oócitos foram avaliados, quanto grau de expansão das células do cumulus, configuração da cromatina, níveis de GSH e mitocôndrias ativas. Não foram observadas diferenças significativas com relação à taxa de expansão de células do cumulus. A percentagem de oócitos em MII foi maior no grupo ECG do que no grupo CON e HCG (P<0,05) e semelhante ao grupo FSH. Em conclusão, a eCG pode ser utilizada em substituição ao FSH na MIV de oócitos ovinos.

Estro e morfologia cervical de ovelhas mestiças de Dorper e Santa Inês submetidas à sincronização do estro / Estrus and cervical morphology of crossbred Dorper and Santa Inês ewes submitted to estrus synchronization

In order to correlate the beginning of estrus to vaginal and cervical morphologies after synchronizationof estrus in ewes. Forty crossbred females of Dorper and Santa Inês were submitted to estrous synchronizationusing CIDR and eCG. After hormonal treatment, we recorded oestrus, morphology of vagina and cervix. Thedata were analyzed using the Analysis of Variance and the Tukey and Chi-square tests. All sheep presentedestrus, beginning at 32.30 ± 0.99 h after removal of the CIDR. There was a predominance of ewes with vaginaland cervical epithelium with pink coloration. At 0 h and 24 h after CIDR removal, 90% and 75% of the ewespresented incomplete crystallization respectively. The maximum crystallization of the cervical mucus occurred at48 hours after withdrawal of the CIDR. Therefore, the histomorphology of the mucosa and cervical mucus variesaccording to the time of estrus of sheep.(AU)

Efeito do óleo essencial de Croton nepetifolius sobre a penetração cervical em ovelhas mestiças de Dorper e Santa Inês / Effect of the essential oil of Croton nepetifolius Baill on cervical penetration in crossbred Dorper and Santa Inês ewes

To evaluate the use of the Croton nepetifolius Baill (OECn) essential oil in cervical penetrability inewe, forty Forty crossbred females of Dorper and Santa Inês were submitted to estrous synchronization anddistributed into four groups (n = 10). A control group and another three groups treated with 200 μg /misoprostol sheep (Misoprostol), 50 μg / mL (OECn50) and 100 μg / mL (OECn100) of OECn oil respectively,before the introduction of an artificial insemination applicator (AI). The depth of cervical penetration and thepassage time of the mandrel of AI were recorded and analyzed using ANOVA and the Tukey and Chi-squaretests. There was no significant difference in cervical penetrability, but the Misoprostol and OECn100 groupspresented a lower penetrability time when compared to the Control group. Therefore, the use of Crotonnepetifolius Baill essential oil facilitates cervical sheep penetration.(AU)

Efeito da melatonina na maturação in vitro de oócitos caprinos / Effect of melatonin on in vitro maturation of goat oocytes

To evaluate the effect of melatonin on in vitro maturation of goat oocytes, 12 females were used asoocyte donor. They were subjected to ovarian stimulation and ovum pickup by laparotomy. Oocytes wereevaluated and allocated into three groups of in vitro maturation: M0 (oocytes matured in TCM199 andsupplements); M10 and M50 (oocytes matured in M0 medium supplemented with 10 μM and 50 μM melatonin,respectively). All groups were subjected to the cell incubator for 24 hours at 38.5°C with a humidifiedatmosphere containing 5% CO2. For data analysis, Chi-square test was used, with a probability of 5% and acorrelation test. In conclusion, melatonin reduced in vitro maturation rates of goat oocytes.(AU)

Atividade estral e ovulatória de cabras submetidas à sincronização do estro com esponjas intravaginais impregnadas com óleos essenciais de Vitex agnus castus L / Estrous and ovulatory activity of goats submitted to estrus synchronization with intravaginal esponges impregnated with essential oils of Vitex agnus castus L

In order to evaluate the estrous and ovulatory activities of goats submitted to estrus synchronizationwith essential oils of Vitex agnus castus L. (OEVAC), 48 Anglo Nubian goats were divided into four groups(n=12) for estrus synchronization: MAP-COM group, 75 µg of D-cloprostenol was intramuscularly (im)administered and one intravaginal sponge impregnated with 60 mg of MAP was deposited and removed fivedays later when it was injected im 300 IU eCG; MAP-ART group with similar treatment of MAP-COM, butintravaginal sponges were manually prepared and impregnated with 60 mg of MAP; in the OEVAC60 andOEVAC120 groups it was performed the same as in MAP-COM, but intravaginal sponges were impregnatedwith 60 mg and 120 mg of OEVAC, respectively. The use of intravaginal sponges impregnated with 60 mg of theOEVAC it is able to synchronize estrus but not the ovulation in goats.(AU)

Atividade estral e ovulatória de cabras submetidas à sincronização do estro com esponjas intravaginais impregnadas com óleos essenciais de Vitex agnus castus L / Estrous and ovulatory activity of goats submitted to estrus synchronization with intravaginal esponges impregnated with essential oils of Vitex agnus castus L

In order to evaluate the estrous and ovulatory activities of goats submitted to estrus synchronizationwith essential oils of Vitex agnus castus L. (OEVAC), 48 Anglo Nubian goats were divided into four groups(n=12) for estrus synchronization: MAP-COM group, 75 µg of D-cloprostenol was intramuscularly (im)administered and one intravaginal sponge impregnated with 60 mg of MAP was deposited and removed fivedays later when it was injected im 300 IU eCG; MAP-ART group with similar treatment of MAP-COM, butintravaginal sponges were manually prepared and impregnated with 60 mg of MAP; in the OEVAC60 andOEVAC120 groups it was performed the same as in MAP-COM, but intravaginal sponges were impregnatedwith 60 mg and 120 mg of OEVAC, respectively. The use of intravaginal sponges impregnated with 60 mg of theOEVAC it is able to synchronize estrus but not the ovulation in goats.

Efeito da melatonina na maturação in vitro de oócitos caprinos / Effect of melatonin on in vitro maturation of goat oocytes

To evaluate the effect of melatonin on in vitro maturation of goat oocytes, 12 females were used asoocyte donor. They were subjected to ovarian stimulation and ovum pickup by laparotomy. Oocytes wereevaluated and allocated into three groups of in vitro maturation: M0 (oocytes matured in TCM199 andsupplements); M10 and M50 (oocytes matured in M0 medium supplemented with 10 μM and 50 μM melatonin,respectively). All groups were subjected to the cell incubator for 24 hours at 38.5°C with a humidifiedatmosphere containing 5% CO2. For data analysis, Chi-square test was used, with a probability of 5% and acorrelation test. In conclusion, melatonin reduced in vitro maturation rates of goat oocytes.

Efeito do óleo essencial de Croton nepetifolius sobre a penetração cervical em ovelhas mestiças de Dorper e Santa Inês / Effect of the essential oil of Croton nepetifolius Baill on cervical penetration in crossbred Dorper and Santa Inês ewes

To evaluate the use of the Croton nepetifolius Baill (OECn) essential oil in cervical penetrability inewe, forty Forty crossbred females of Dorper and Santa Inês were submitted to estrous synchronization anddistributed into four groups (n = 10). A control group and another three groups treated with 200 μg /misoprostol sheep (Misoprostol), 50 μg / mL (OECn50) and 100 μg / mL (OECn100) of OECn oil respectively,before the introduction of an artificial insemination applicator (AI). The depth of cervical penetration and thepassage time of the mandrel of AI were recorded and analyzed using ANOVA and the Tukey and Chi-squaretests. There was no significant difference in cervical penetrability, but the Misoprostol and OECn100 groupspresented a lower penetrability time when compared to the Control group. Therefore, the use of Crotonnepetifolius Baill essential oil facilitates cervical sheep penetration.

Estro e morfologia cervical de ovelhas mestiças de Dorper e Santa Inês submetidas à sincronização do estro / Estrus and cervical morphology of crossbred Dorper and Santa Inês ewes submitted to estrus synchronization

In order to correlate the beginning of estrus to vaginal and cervical morphologies after synchronizationof estrus in ewes. Forty crossbred females of Dorper and Santa Inês were submitted to estrous synchronizationusing CIDR and eCG. After hormonal treatment, we recorded oestrus, morphology of vagina and cervix. Thedata were analyzed using the Analysis of Variance and the Tukey and Chi-square tests. All sheep presentedestrus, beginning at 32.30 ± 0.99 h after removal of the CIDR. There was a predominance of ewes with vaginaland cervical epithelium with pink coloration. At 0 h and 24 h after CIDR removal, 90% and 75% of the ewespresented incomplete crystallization respectively. The maximum crystallization of the cervical mucus occurred at48 hours after withdrawal of the CIDR. Therefore, the histomorphology of the mucosa and cervical mucus variesaccording to the time of estrus of sheep.