Efeito do cilostazol na produção in vitro de ovinos / Effect of cilostazol on in vitro production of sheep / Efecto del cilostazol en la producción in vitro de ovejas

O objetivo deste estudo foi avaliar o efeito da inclusão de cilostazol no meio de maturação in vitro de oócitos sobre produção in vitro de embriões ovinos. Para isso, foram realizadas colheitas de oócitos oriundos de ovários obtidos em abatedouro por meio do método de aspiração folicular com bomba de vácuo. Os oócitos foram divididos em quatro grupos de maturação: grupo CON, onde os complexos cumulus oócitos foram imersos em TCM-199, suplementado com 500 UI de penicilina, 0,5 mg de estreptomicina, 1,25 µg de anfotericina, 0,2 mM de piruvato de sódio, 10% (v/v) de soro fetal bovino (SFB), 10 ng/mL de fator de crescimento epidérmico (EGF), 10 ug/m de FSH, 10 µg/mL de LH, 10 ug/mL de estradiol e 100 µM de cisteamina; e nos grupos CILO0,3; CILO1 e CILO10, os oócitos foram maturados no meio do grupo CON, mas sem a adição de cisteamina e suplementado com as concentrações de 0,3; 1 e 10 µM, respectivamente. Após 24h, os oócitos foram avaliados quanto a presença ou não de células do cumulus e quanto ao grau de expansão e destinados à fecundação in vitro, em meio FIV, juntamente com espermatozoides. Após a FIV, os presumíveis zigotos seguiram para o cultivo in vitro. Foram avaliadas clivagens no dia 2, sendo dia 0 o dia do início do CIV. Os resultados foram expressos em porcentagem e as variáveis de expansão das células do cumulus e número de estruturas clivadas foram comparadas por meio do teste qui-quadrado do software Epi Info (Epi Info 7.2.5, Atlanta, GA, EUA, 2021). Os resultados foram considerados significativos quando P<0,05. Em relação à expansão das células do cumulus, todos os grupos apresentaram 100% de expansão. Não houve diferenças significativas quanto ao grau de expansão das células do cumulus entre os grupos suplementados com cilostazol e cisteamina (P>0,05), assim como não houve diferenças significativas entre as taxas de clivagem entre os grupos suplementados com cilostazol e cisteamina (P > 0,05).

The objective of this study was to evaluate the effect of including cilostazol in the in vitro maturation medium of oocytes on the in vitro production of sheep embryos. Oocytes were collected from ovaries obtained from a slaughterhouse by follicular aspiration with a vacum pump. The oocytes were divided into four maturation groups: the CON group, where the cumulus-oocyte complexes were immersed in TCM-199 supplemented with 500 IU of penicillin, 0.5 mg of streptomycin, 1.25 µg of amphotericin, 0.2 mM of sodium pyruvate, 10% (v/v) fetal bovine serum (FBS), 10 ng/mL of epidermal growth factor (EGF), 10 µg/mL of FSH, 10 µg/mL of LH, 10 µg/mL of estradiol, and 100 µM of cysteamine; and in the CILO0.3, CILO1, and CILO10 groups, the oocytes were matured in the CON group medium without the addition of cysteamine and supplemented with concentrations of 0.3, 1, and 10 µM of cilostazol, respectively. After 24 hours, the oocytes were evaluated for the presence or absence of cumulus cells and the degree of expansion and then subjected to in vitro fertilization (IVF) with sperm in FIV medium. After IVF, the presumptive zygotes were cultured in vitro. Cleavage was evaluated on day 2, with day 0 being the start of IVF. Results were expressed as a percentage, and variables such as cumulus cell expansion and the number of cleaved structures were compared using the chi-square test in the Epi Info software (Epi Info 7.2.5, Atlanta, GA, USA, 2021). Results were considered significant when P < 0.05. All groups showed 100% cumulus cell expansion, and there were no significant differences in cumulus cell expansion degree between the cilostazol- and cysteamine-supplemented groups (P > 0.05), as well as no significant differences in cleavage rates between the cilostazol- and cysteamine-supplemented groups (P > 0.05).

El objetivo de este estudio fue evaluar el efecto de la inclusión de cilostazol en el medio de maduración in vitro de ovocitos sobre la producción in vitro de embriones ovinos. Para ello, se realizaron recolecciones de ovocitos provenientes de ovarios obtenidos en un matadero mediante el método de aspiración folicular con bomba de vacío. Los ovocitos se dividieron em cuatro grupos de maduración: grupo CON, donde los complejos cúmulus ovocitos se sumergieron en TCM-199, suplementado con 500 UI de penicilina, 0,5 mg de estreptomicina, 1,25 ug de anfotericina, 0,2 mM de piruvato de sodio, 10% (v/v) de suero fetal bovino (SFB), 10 ng/mL de factor de crecimiento epidérmico (EGF), 10 ug/m de FSH, 10 µg/mL de LH, 10 µg/mL de estradiol y 100 µM de cisteamina; y en los grupos CILO0,3; CILO1 y CILO10, los ovocitos se maduraron en el medio del grupo CON, pero sin la adición de cisteamina y suplementado con las concentraciones de 0,3; 1 y 10 µM, respectivamente. Después de 24 horas, los ovocitos se evaluaron en cuanto a la presencia o no de células del cúmulus y em cuanto al grado de expansión y se destinaron a la fecundación in vitro, en medio FIV, junto con espermatozoides. Después de la FIV, los presuntos cigotos siguieron para el cultivo in vitro. Se evaluaron las clivajes en el día 2, siendo el día 0 el día del início del CIV. Los resultados se expresaron en porcentaje y las variables de expansión de las células del cúmulos y número de estructuras clivadas se compararon mediante la prueba del chi-cuadrado del software Epi Info (Epi Info 7.2.5, Atlanta, GA, EE. UU., 2021). Los resultados se consideraron significativos cuando P < 0,05. En relación a la expansión de las células del cúmulus, todos los grupos presentaron el 100% de expansión. No hubo diferencias significativas en cuanto al grado de expansión de las células del cúmulus entre los grupos suplementados con cilostazol y cisteamina (P > 0.05), así como no hubo diferencias significativas entre las tasas de clivaje entre los grupos suplementados con cilostazol y cisteamina (P>0,05).

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.

Uso de hCG e deslorelina na indução de ovulação de éguas cíclicas no Agreste de Pernambuco / Use of hCG and deslorelin in mares ovulation induction in Agreste of Pernambuco

The experiment aimed to compare the efficiency of deslorelin and hCG to induce ovulation Quarter Horsemares. This experiment was conducted at the Center for Equine Pernambuco - CEPE, in the town of Gravatá-PE, in February 2013. The animals were divided in Group 1 (n=10) received, intramuscular, Deslorelin 1000mg while Group 2 (n = 10) received, Intravenous, 5000 IU of hCG. 24 hours after administration of theovulation inducing were inseminated with semen from a stallion. After 15 days of AI, the pregnancy diagnosiswas made. The data from this study were analyzed using the statistical package SAS, (P 0.05). The drugs tested successfully induce ovulation.

Uso de hCG e deslorelina na indução de ovulação de éguas cíclicas no Agreste de Pernambuco / Use of hCG and deslorelin in mares ovulation induction in Agreste of Pernambuco

The experiment aimed to compare the efficiency of deslorelin and hCG to induce ovulation Quarter Horsemares. This experiment was conducted at the Center for Equine Pernambuco - CEPE, in the town of Gravatá-PE, in February 2013. The animals were divided in Group 1 (n=10) received, intramuscular, Deslorelin 1000mg while Group 2 (n = 10) received, Intravenous, 5000 IU of hCG. 24 hours after administration of theovulation inducing were inseminated with semen from a stallion. After 15 days of AI, the pregnancy diagnosiswas made. The data from this study were analyzed using the statistical package SAS, (P < 0.05). The fertilityrate found was 80.0% in group 1 and 70% in group 2 (P > 0.05). The drugs tested successfully induce ovulation.(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 estimulação hormonal ovariana e polivinilpirrolidona sobre a maturação in vitro de oócitos de cabras Anglo Nubiana / Effect of hormonal ovarian stimulation and polyvinylpyrrolidone on in vitro maturation of oocytes of Anglo Nubian goats

To evaluate the effect of hormonal stimulation treatments with PVP on the production and maturationof COCs, 27 goats received intravaginal devices (CIDR) for 10 days and 125 µg cloprostenol on day 8 oftreatment. The goats were distributed in the following groups: FSH-eCG, 300 IU of eCG and 70 mg pFSH, 36 hprior to the withdrawal of CIDR; FSH, receiving 180 mg (40/40; 35/35, 30 mg) at 12 h intervals; FSH-PVP10and FSH-PVP40 receiving 70 mg FSH dissolved in PVP 10,000 MW (24 hours before CIDR) and PVP MW40,000 (48 hours before CIDR), respectively. After follicular aspiration, COCs of grade I and II were submittedto IVM. Recovery rate from the FSH-PVP10 was higher (73.33%) than FSH-PVP40 (44.26%) and FSH-eCG(27.14%). COCs with grades I and II, were more frequently observed (P < 0.05) in FSH-PVP40 than FSH-eCG.FSH-PVP40 is an alternative to recovery and mature oocytes in vitro.

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.

Resposta folicular após estimulação ovariana com gonadotrofinas e polivinilpirrolidona em cabras Anglo Nubiana / Follicular response after ovarian stimulation with gonadotropins and polyvinylpyrrolidone in Anglo Nubian goats

To evaluate the effect of different hormonal stimulation treatments with PVP on ovarian folliculardevelopment, 27 goats received intravaginal devices (CIDR) for 10 days and 125 µg cloprostenol on day 8 oftreatment. The goats were distributed in the following groups: G1 (FSH-eCG), 300 IU of eCG and 70 mg pFSH,36 h prior to the withdrawal of CIDR; G2 (FSH), receiving 180 mg (40/40; 35/35, 30 mg) at 12 h intervals; G3(FSH-PVP10), receiving 70 mg FSH dissolved in PVP 10,000 MW, 24 hours before CIDR; and G4 (FSHPVP40),70 mg of FSH dissolved in PVP MW 40,000, 48 hours before CIDR. The follicles with diameter ≥ 2 mmwere counted, measured and classified. FSH-PVP 40 had a greater number of small follicles than FSH (P<0.05)and FSH showed greater number of large follicles (P < 0.05). In conclusion, treatment with FSH-PVP40 is analternative to stimulate ovaries in goats.

Uso do óleo essencial de Croton nepetaefolius Baill (Marmeleiro vermelho) na colheita transcervical de embriões de ovelhas Rabo Largo / Use of the essential oil of Croton nepetaefolius Baill (red quince) in transcervical embryo recovery of Rabo Largo ewes

With the objective of evaluating the Croton nepetaefolius Baill in the transcervical harvest of ovineembryos, we used twelve sheeps and two rams of Rabo Largo breed. After estrus synchronization andsuperovulation, the females were submitted to the embryonic transcervical harvest, being randomly distributedinto three groups (n = 4). The PGF2α group received 75 μg of cloprostenol (IM), twenty four hours beforeharvest; and groups OECn100 and OECn300 received in the background of the vaginal sac, five minutes beforeharvest, 100 μg / mL and 300 μg / mL of the oil, respectively. We used ANOVA and the Tukey test for the resultsanalysis, at a probability of 5%. There was no significant difference for all parameters, between the treatmentgroups (P > 0.05). Croton nepetaefolius Baill essential oil did not facilitate cervical transposition of allharvested animals and did not result in embryo recovery.

Uso do óleo essencial de Croton nepetaefolius Baill (Marmeleiro vermelho) na colheita transcervical de embriões de ovelhas Rabo Largo / Use of the essential oil of Croton nepetaefolius Baill (red quince) in transcervical embryo recovery of Rabo Largo ewes

With the objective of evaluating the Croton nepetaefolius Baill in the transcervical harvest of ovineembryos, we used twelve sheeps and two rams of Rabo Largo breed. After estrus synchronization andsuperovulation, the females were submitted to the embryonic transcervical harvest, being randomly distributedinto three groups (n = 4). The PGF2α group received 75 μg of cloprostenol (IM), twenty four hours beforeharvest; and groups OECn100 and OECn300 received in the background of the vaginal sac, five minutes beforeharvest, 100 μg / mL and 300 μg / mL of the oil, respectively. We used ANOVA and the Tukey test for the resultsanalysis, at a probability of 5%. There was no significant difference for all parameters, between the treatmentgroups (P > 0.05). Croton nepetaefolius Baill essential oil did not facilitate cervical transposition of allharvested animals and did not result in embryo recovery.(AU)

Resposta folicular após estimulação ovariana com gonadotrofinas e polivinilpirrolidona em cabras Anglo Nubiana / Follicular response after ovarian stimulation with gonadotropins and polyvinylpyrrolidone in Anglo Nubian goats

To evaluate the effect of different hormonal stimulation treatments with PVP on ovarian folliculardevelopment, 27 goats received intravaginal devices (CIDR) for 10 days and 125 µg cloprostenol on day 8 oftreatment. The goats were distributed in the following groups: G1 (FSH-eCG), 300 IU of eCG and 70 mg pFSH,36 h prior to the withdrawal of CIDR; G2 (FSH), receiving 180 mg (40/40; 35/35, 30 mg) at 12 h intervals; G3(FSH-PVP10), receiving 70 mg FSH dissolved in PVP 10,000 MW, 24 hours before CIDR; and G4 (FSHPVP40),70 mg of FSH dissolved in PVP MW 40,000, 48 hours before CIDR. The follicles with diameter ≥ 2 mmwere counted, measured and classified. FSH-PVP 40 had a greater number of small follicles than FSH (P<0.05)and FSH showed greater number of large follicles (P < 0.05). In conclusion, treatment with FSH-PVP40 is analternative to stimulate ovaries in goats.(AU)

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)

Efeito da estimulação hormonal ovariana e polivinilpirrolidona sobre a maturação in vitro de oócitos de cabras Anglo Nubiana / Effect of hormonal ovarian stimulation and polyvinylpyrrolidone on in vitro maturation of oocytes of Anglo Nubian goats

To evaluate the effect of hormonal stimulation treatments with PVP on the production and maturationof COCs, 27 goats received intravaginal devices (CIDR) for 10 days and 125 µg cloprostenol on day 8 oftreatment. The goats were distributed in the following groups: FSH-eCG, 300 IU of eCG and 70 mg pFSH, 36 hprior to the withdrawal of CIDR; FSH, receiving 180 mg (40/40; 35/35, 30 mg) at 12 h intervals; FSH-PVP10and FSH-PVP40 receiving 70 mg FSH dissolved in PVP 10,000 MW (24 hours before CIDR) and PVP MW40,000 (48 hours before CIDR), respectively. After follicular aspiration, COCs of grade I and II were submittedto IVM. Recovery rate from the FSH-PVP10 was higher (73.33%) than FSH-PVP40 (44.26%) and FSH-eCG(27.14%). COCs with grades I and II, were more frequently observed (P < 0.05) in FSH-PVP40 than FSH-eCG.FSH-PVP40 is an alternative to recovery and mature oocytes in vitro.(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)

Effect of pFSH dose reduction on in vivo embryo production in Dorper ewes / Efeito da redução da dose de pFSH na produção in vivo de embriões em ovelhas Dorper

To evaluate the effect of pFSH dose on the in vivo embryo production of Dorper ewes in the semi-arid northeast of Brazil, 40 sheep females were distributed into two groups of 20 animals that received intravaginal CIDR for 14 days, and two days before device removal, they received one of the following treatments: in the FSH200 group, the ewes received 200 mg of pFSH; and in the FSH128 group, the ewes received a total of 128 mg in decreasing doses every 12 h. Beginning 12 h after the conclusion of the treatments, estrus detection was performed every four hours using two Dorper rams of proven fertility. The ewes were mated at estrus onset and 24 hours later. Seven days after intravaginal device removal, the superovulatory response was evaluated, and embryo collection was performed using the laparotomy method. The recovered flushings were subjected to embryo searches under a stereomicroscope and classified according to their qualities. Analyses of variance (ANOVAs) and LSD tests were used to compare the different parameters. The data expressed as percentages were analysed by chi-square test. The ovulation rate was higher in the FSH200 group, which had 16.3 ± 0.3 corpora lutea (CL), than in the FSH128 group, which had 11.3 ± 0.3 CL (P 0.05). However, higher fertilization rate (83.6% vs. 62.4%) and higher transferable (86.0% vs. 71.6%) and freezable (67.9% vs. 40.8%) embryo rates were observedin the FSH 128 group compared with the group that received 200 mg. Furthermore, no significant differences in the remaining parameters were observed between the experimental groups (P>0.05), demonstrating that pFSH dose reduction promoted a greater production of freezable and transferableembryos in Dorper ewes subjected to MOET.

Com o objetivo de avaliar o efeito da redução de pFSH na produção in vivo de embriões de ovelhas da raça Dorper exploradas no semiárido do Nordeste do Brasil, 40 fêmeas ovinas foram distribuídas em dois grupos com 20 animais, os quais receberam um CIDR intravaginal por 14 dias e dois dias antes da remoção do dispositivo receberam um tratamento superovulatório proposto: grupo FSH200 onde as ovelhas receberam 200 mg de pFSH, enquanto o grupo FSH128 totalizou 128 mg em doses decrescentes intervaladas por 12 horas. Doze horas após o final do tratamento, foi realizada a detecção de estro, a cada quatro horas, utilizando dois carneiros Dorper, de fertilidade comprovada. As ovelhas foram cobertas no início do estro e 24 horas depois. Sete dias após a retirada do dispositivo intravaginal, foi realizada a avaliação da resposta superovulatória e a colheita de embriões pelo método de laparotomia. O lavado recuperado foi submetido à procura de embriões em estereomicroscópio e classificados de acordo com sua qualidade. Para comparação dos diversos parâmetros, foi utilizada a Análise de Variância (ANOVA), seguida da realização do teste LSD. Os dados expressos em porcentagem foram submetidos ao Teste de Qui-quadrado. A taxa de ovulação foi superior no grupo FSH200 com 16,3 ± 0,3 corpos lúteos (CL) quando comparado ao grupo FSH128 com de 11,3 ± 0,3 CL (P 0,05). Entretanto, foi verificada uma maior taxa de fecundação (83,6% vs. 62,4%) e uma maior taxa de embriões transferíveis (86,0% vs.71,6%) e congeláveis (67,9% vs. 40,8%) no grupo FSH 128 quando comparado ao grupo que recebeu200 mg. Além disso, não foi observada qualquer diferença significativa entre os grupos experimentais, para os demais parâmetros avaliados demonstrando que a redução da dose de pFSH promove maior produção de embriões congeláveis e transferíveis em ovinos Dorper submetidos à MOTE.

Addition of sucrose on cryoprotectant solution of vitrification enhances the quality of Dorper sheep embryos produced in vivo / Adição de sacarose na solução crioprotetora de vitrificação melhora a qualidade de embriões ovinos Dorper produzidos in vivo

This study aimed to evaluate the efficacy of adding sucrose in vitrification solution of ovine embryos produced in vivo. Forty Dorper ewes were selected and superovulated. Immediately prior to the embryo collection by laparotomy, a laparoscopy was performed to verify the superovulatory response. The recovered flushing was followed by embryo evaluation and embryos were divided in two experimental groups where embryos from Control group were submitted to a traditional vitrification protocol and embryos from Sucrose group to a modified vitrification protocol with sucrose. After warming, embryos were again divided regarding cryoprotectant removal (Indirect) or not (Direct). The embryo quality was classified as embryos of degrees I (excellent or good), II (regular), III (poor) and IV (dead or degenerate). It was also verified the homogeneity of mass, occurrence of embryonic mass retraction and rupture of pellucid zone. The results were expressed as percentages and were subjected to Chi-square test with P 0.05. The embryos vitrified in the presence of sucrose had lower proportions of lower-quality embryos after warming (22.20 vs. 44.50%), higher percentages of homogeneous embryos after warming (63.89 vs. 38.89 %) while concerning other parameters there was no difference between these groups. It can be concluded that the addition of 0.4 M sucrose during vitrification improves the embryo quality.

Este estudo objetivou avaliar a eficácia da adição de sacarose na solução de vitrificação de embriões ovinos produzidos in vivo. Foram selecionadas 40 ovelhas da raça Dorper as quais foram superovuladas. Imediatamente antes da colheita de embriões por laparotomia, uma laparoscopia foi realizada para verificar a resposta superovulatória. O lavado recuperado foi submetido à procura e avaliação de embriões e estes foram divididos em dois grupos experimentais, onde os embriões do Grupo Controle foram submetidos ao protocolo tradicional de vitrificação e os embriões do Grupo Sacarose a um protocolo modificado de vitrificação com sacarose. Após a descongelação, os embriões foram novamente divididos considerando a remoção (Indireto) ou não (Direto) do crioprotetor. A qualidade embrionária foi classificada como embriões de graus I (excelente ou bom), II (regular), III (pobre) e IV (morto ou degenerado). Foi também verificada a homogeneidade da massa, ocorrência de retração da massa e ruptura de zona pelúcida. Os resultados foram expressos em porcentagem e foram submetidos ao teste do Qui-quadrado com P < 0.05. Os embriões vitrificados na presença de sacarose apresentaram menores proporções de embriões de menor qualidade após a descongelação (22,20 vs. 44,50%), e maiores percentuais de embriões homogêneos após a descongelação (63,89 vs. 38,89%) enquanto com relação aos demais parâmetros não existiram diferenças entre grupos. Pode-se concluir que a adição de 0,4 M de sacarose durante os procedimentos de vitrificação e descongelação beneficia a qualidade embrionária.