RESUMO
The aims of the present study were to evaluate the protective effects of gallic acid against doxorubicin-induced ovarian toxicity in mice, and to verify the possible involvement of PI3K and mTOR signaling pathway members (PTEN, Akt, FOXO3a and rpS6) in the gallic acid protective actions. Mice were pretreated with NaCl (0.15 M, p.o.) (control and doxorubicin groups) or gallic acid (50, 100 or 200 mg/kg body weight, p.o.) once daily, for 5 days, and on the third day of treatment, after 1 h of treatment administration, the mice received saline solution (i.p.) (control group) or doxorubicin (10 mg/kg of body weight, i.p.). Next, the ovaries were harvested for histological (follicular morphology and activation), fluorescence (GSH and mitochondrial activity), and immunohistochemical (PCNA, cleaved caspase-3, TNF-α, p-PTEN, Akt, p-Akt, p-rpS6 and p-FOXO3a) analyses. The results showed that cotreatment with 50 mg/kg gallic acid plus doxorubicin preserved the percentage of normal follicles and cell proliferation, reduced the percentage of cleaved caspase-3 follicles, prevented inflammation, and increased GSH concentrations and mitochondrial activity compared to doxorubicin treatment alone. Furthermore, cotreatment 50 mg/kg gallic acid plus doxorrubicin increased expression of Akt, p-Akt, p-rpS6 and p-FOXO3a compared to the doxorubicin alone. In conclusion, 50 mg/kg gallic acid protects the mouse ovary against doxorubicin-induced damage by improving GSH concentrations and mitochondrial activity and cellular proliferation, inhibiting inflammation and apoptosis, and regulating PI3K and mTOR signaling pathway.
Assuntos
Ovário , Proteínas Proto-Oncogênicas c-akt , Feminino , Camundongos , Animais , Proteínas Proto-Oncogênicas c-akt/metabolismo , Folículo Ovariano , Caspase 3/metabolismo , Ácido Gálico/farmacologia , Ácido Gálico/uso terapêutico , Serina-Treonina Quinases TOR/metabolismo , Doxorrubicina/toxicidade , Fosfatidilinositol 3-Quinases , Inflamação/metabolismo , ApoptoseRESUMO
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.
Assuntos
Animais , Ovinos , Técnicas de Maturação in Vitro de Oócitos/veterinária , Hormônio Foliculoestimulante , Gonadotropina CoriônicaRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Many studies are performed with the aerial parts of Cannabis sativa L. (Cannabaceae). However, roots remain poorly studied, despite citations in the scientific literature. The C. sativa roots are indicated for the treatment of pain, inflammation, fever, among other health problems. AIM OF THE STUDY: This study aimed to evaluate the antinociceptive, antipyretic, antiasthmatic, and spasmolytic activities of C. sativa roots in experimental models using mice and rats. MATERIAL AND METHODS: The chemical composition of the aqueous extract of C. sativa roots (AECsR) was evaluated by LC-MS. The antinociceptive activity was assessed in mice by the induction of writhing with acetic acid, paw licking with formalin, and reactivity in the hot plate test. Fever was induced by the administration of a suspension of Saccharomyces cerevisiae in young rats. The asthmatic activity was performed with ovalbumin (OVA)-immunized mice with cellular and histological analysis. Finally, the spasmolytic activity was performed using mice isolated trachea. For in vivo studies, the doses were 12.5, 25, or 50 mg/kg whereas for in vitro, the concentration of AECsR was 729 µg/mL. RESULTS: From the LC-MS data, we identified p-coumaroyltyramine, feruloyltyramine canabissativine in AECsR. The extract promoted a reduction of writhing in all tested doses (12.5, 25, or 50 mg/kg). Similarly, it reduced the pain in the formalin test at doses of 12.5 and 50 mg/kg (first phase) and 12.5 and 25 mg/kg (second phase). In the hot plate test, the doses of 12.5, 25, and 50 mg/kg promoted antinociceptive effect at different times, and the lowest dose maintained its action in the analyzes performed at 60, 90, and 120 min after administration. The anti-inflammatory activity of AECsR was observed in the mouse model of asthma, reducing the total leukocyte count in the bronchoalveolar fluid (BALF) at a dose of 25 mg/kg, as well as reducing eosinophilia in all tested doses (12.5, 25, and 50 mg/kg). Histological analysis of lungs stained with H&E and PAS showed a reduction in the number of inflammatory cells in the perivascular and peribronchial region, as well as reduced mucus production. CONCLUSION: The results suggest that AECsR promotes pain control, either by a central or inflammatory mechanism, and has antiasthmatic activity. However, there was no antipyretic or spasmolytic effect.
Assuntos
Analgésicos/farmacologia , Antiasmáticos/farmacologia , Cannabis/química , Extratos Vegetais/farmacologia , Analgésicos/administração & dosagem , Analgésicos/isolamento & purificação , Animais , Antiasmáticos/administração & dosagem , Antiasmáticos/isolamento & purificação , Antipiréticos/administração & dosagem , Antipiréticos/isolamento & purificação , Antipiréticos/farmacologia , Brasil , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Febre/tratamento farmacológico , Inflamação/tratamento farmacológico , Masculino , Camundongos , Dor/tratamento farmacológico , Parassimpatolíticos/administração & dosagem , Parassimpatolíticos/isolamento & purificação , Parassimpatolíticos/farmacologia , Extratos Vegetais/administração & dosagem , Raízes de Plantas , Ratos , Ratos WistarRESUMO
The present study evaluated the effects of protocatechuic acid (PCA) after cisplatin-induced ovarian toxicity in mice and if PTEN and FOXO3a proteins are involved in PCA action. The mice were divided into five experimental groups (five animals per group) and treated once a day for 3 days as follows: (1) the control group was pretreated with oral administration (o.p.) of saline solution, followed by an intraperitoneal (i.p.) injection of saline solution. The other groups were pretreated (o.p.) with (2) saline solution (cisplatin group), (3) N-acetylcysteine (150 mg/kg of body weight), or with (4) 20 or (5) 50 mg/kg body weight of PCA, followed by 5 mg/kg body weight (i.p.) of cisplatin. Next, the ovaries were destined to histological (morphology and activation), immunohistochemical (PCNA and cleaved caspase-3 expression), and fluorescence (reactive oxygen species [ROS], glutathione [GSH], and active mitochondria levels) analyses. Moreover, the immunoreactivity for p-PTEN and p-FOXO3a was evaluated to investigate a potential mechanism by which PCA could prevent the cisplatin-induced ovarian damage. Pretreatment with N-acetylcysteine or 20 mg/kg PCA before cisplatin preserved the percentage of normal follicles and cell proliferation as observed in the control, reduced apoptosis and ROS levels, and showed higher active mitochondria and GSH levels than the cisplatin treatment (P < 0.05). Moreover, pretreatment with 20 mg/kg PCA decreased cisplatin-induced p-PTEN and increased (P < 0.05) nuclear export of p-FOXO3a. In conclusion, PCA at 20 mg/kg reduced apoptosis, maintained cell proliferation and mitochondrial function, reduced ROS production, and increased GSH expression likely through the involvement of PTEN and FOXO3a proteins.
Assuntos
Proteína Forkhead Box O3/metabolismo , Hidroxibenzoatos/farmacologia , Doenças Ovarianas/prevenção & controle , Ovário/efeitos dos fármacos , PTEN Fosfo-Hidrolase/metabolismo , Substâncias Protetoras/farmacologia , Animais , Apoptose/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Cisplatino , Modelos Animais de Doenças , Feminino , Glutationa/metabolismo , Camundongos , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Mitocôndrias/patologia , Doenças Ovarianas/induzido quimicamente , Doenças Ovarianas/enzimologia , Doenças Ovarianas/patologia , Ovário/metabolismo , Ovário/patologia , Fosforilação , Espécies Reativas de Oxigênio/metabolismoRESUMO
The objective of this study was to test the efficiency of powdered coconut water (ACP-406®) base-medium without or with the addition of supplements on in vitro culture of isolated goat secondary follicles. Follicles were cultured for 18 days in α-MEM or in ACP-406®, both without supplements (referred to as α-MEM and ACP, respectively), or both supplemented with BSA, insulin, transferrin, selenium, glutamine, hypoxanthine, and ascorbic acid (referred to as α-MEM+ and ACP+). Follicular morphology, antrum formation, follicular and oocyte diameter, levels of glutathione (GSH), and chromatin configuration after in vitro maturation were evaluated. At the end of culture, ACP-406® base-medium (without or with supplements) showed a higher (P < 0.05) percentage of normal follicles than α-MEM (without or with supplements). Antrum formation was similar among α-MEM+, ACP and ACP+, and significantly higher than α-MEM without supplements. The follicular diameter was greater in ACP+ than α-MEM, and similar to other treatments. Moreover, fully and daily grown rates were higher (P < 0.05) in ACP-406® base-medium (without or with supplements) than α-MEM (without or with supplements). Levels of GSH were similar between ACP+ and α-MEM+ treatments. Both ACP+ and α-MEM+ allowed meiotic resumption without a significant difference between the two groups. In conclusion, supplemented ACP-406® base-medium maintained follicular survival and promoted the development as well as meiotic resumption of isolated goat secondary follicles cultured in vitro for 18 days.
RESUMO
The objective of this study was to test the efficiency of powdered coconut water (ACP-406®) base-medium without or with the addition of supplements on in vitro culture of isolated goat secondary follicles. Follicles were cultured for 18 days in α-MEM or in ACP-406®, both without supplements (referred to as α-MEM and ACP, respectively), or both supplemented with BSA, insulin, transferrin, selenium, glutamine, hypoxanthine, and ascorbic acid (referred to as α-MEM+ and ACP+). Follicular morphology, antrum formation, follicular and oocyte diameter, levels of glutathione (GSH), and chromatin configuration after in vitro maturation were evaluated. At the end of culture, ACP-406® base-medium (without or with supplements) showed a higher (P < 0.05) percentage of normal follicles than α-MEM (without or with supplements). Antrum formation was similar among α-MEM+, ACP and ACP+, and significantly higher than α-MEM without supplements. The follicular diameter was greater in ACP+ than α-MEM, and similar to other treatments. Moreover, fully and daily grown rates were higher (P < 0.05) in ACP-406® base-medium (without or with supplements) than α-MEM (without or with supplements). Levels of GSH were similar between ACP+ and α-MEM+ treatments. Both ACP+ and α-MEM+ allowed meiotic resumption without a significant difference between the two groups. In conclusion, supplemented ACP-406® base-medium maintained follicular survival and promoted the development as well as meiotic resumption of isolated goat secondary follicles cultured in vitro for 18 days.
Assuntos
Feminino , Animais , Alimentos de Coco , Cabras/embriologia , Cabras/fisiologia , Folículo Ovariano/crescimento & desenvolvimento , Folículo Ovariano/química , Técnicas de Maturação in Vitro de Oócitos/veterináriaRESUMO
The objective of this study was to test the efficiency of powdered coconut water (ACP-406®) base-medium without or with the addition of supplements on in vitro culture of isolated goat secondary follicles. Follicles were cultured for 18 days in α-MEM or in ACP-406®, both without supplements (referred to as α-MEM and ACP, respectively), or both supplemented with BSA, insulin, transferrin, selenium, glutamine, hypoxanthine, and ascorbic acid (referred to as α-MEM+ and ACP+). Follicular morphology, antrum formation, follicular and oocyte diameter, levels of glutathione (GSH), and chromatin configuration after in vitro maturation were evaluated. At the end of culture, ACP-406® base-medium (without or with supplements) showed a higher (P < 0.05) percentage of normal follicles than α-MEM (without or with supplements). Antrum formation was similar among α-MEM+, ACP and ACP+, and significantly higher than α-MEM without supplements. The follicular diameter was greater in ACP+ than α-MEM, and similar to other treatments. Moreover, fully and daily grown rates were higher (P < 0.05) in ACP-406® base-medium (without or with supplements) than α-MEM (without or with supplements). Levels of GSH were similar between ACP+ and α-MEM+ treatments. Both ACP+ and α-MEM+ allowed meiotic resumption without a significant difference between the two groups. In conclusion, supplemented ACP-406® base-medium maintained follicular survival and promoted the development as well as meiotic resumption of isolated goat secondary follicles cultured in vitro for 18 days.(AU)
Assuntos
Animais , Feminino , Cabras/embriologia , Cabras/fisiologia , Folículo Ovariano/química , Folículo Ovariano/crescimento & desenvolvimento , Alimentos de Coco , Técnicas de Maturação in Vitro de Oócitos/veterináriaRESUMO
Forage production in the Brazilian semiarid region can be enhanced by irrigation and shading to maximize water use. We evaluated the productivity and morphological characteristics of buffel grass (Cenchrus ciliaris) intercropped with butterfly pea (Clitoria ternatea) under shade conditions using an irrigated area with an artificial shading system (26, 39, 55%, or full sunlight). Butterfly pea was implemented in half of the land parcels in a randomized block using a split plot design with three replications. One hundred twenty days after planting was designated as the uniformity cut and the cutting was performed every 45 days for four consecutive cycles. The morphological analysis was performed during the first and fourth cycle, and the herbage mass was determined. The shadow had a quadratic effect on the leaf appearance rate in the first cycle, while shading had a quadratic deviation effect on the leaf elongation rate in the second cycle and an increased linear effect on the length of the sheet in the first cycle. Shading had a quadratic effect on the herbage mass in the third cycle and decreased linearity in the fourth cycle. The highest percentage of buffel grass was found in the single system in the first and second cycles. In addition, we observed a decreased linear effect of shade in the first cycle and the quadratic deviation effect of shade in the second cycle. A quadratic effect of shade was observed in the third and fourth cycles. The survival of the butterfly pea had a quadratic effect with a minimum rate level of 21% shading. The crude protein content showed a system effect in four cycles with higher crude protein in the intercropping system in the first and second cycles. There was also a quadratic effect of shade in the third cycle and linear increase in the fourth cycle. In conclusion, shading increased the leaf appearance rates and elongation at intermediate levels. In addition, a high level of shading increased the leaf length and survival of butterfly pea while it decreased the mass of the forage produced.
A produção de forragem na região semiárida brasileira pode ser intensificada com a irrigação e o sombreamento, maximizando o uso da água. Com o objetivo de avaliar as características produtivas e morfológicas do Capim Buffel (Cenchrus ciliaris) consorciado com Cunhã (Clitoria ternatea), sob sombreamento, foi utilizada uma área irrigada com sistema de sombreamento artificial (26, 39, 55% ou pleno sol). Foi implantada Cunhã em metade das parcelas em delineamento de blocos casualizados em esquemas de parcelas subdivididas com três repetições. Após 120 dias foi realizado o corte de uniformização e a cada 45 dias foram realizados os cortes por quatro ciclos consecutivos, sendo realizadas análises morfológicas e determinada a massa de forragem no 1.º e 4º ciclos. Houve efeito quadrático para sombra na taxa de aparecimento foliar no 1º ciclo, efeito de desvio da quadrática para sombreamento na taxa de alongamento foliar no 2º ciclo e efeito linear crescente para sombreamento no comprimento da folha no 1º ciclo. A massa de forragem apresentou efeito quadrático para sombreamento no 3º ciclo e efeito linear decrescente no 4º ciclo. O maior percentual de Capim Buffel foi encontrado no sistema solteiro no 1º e 2º ciclos. Foi observado efeito linear decrescente para sombra no 1º ciclo e de desvio da quadrática para sombra no 2º ciclo. Já no 3º e 4º ciclos houve efeito quadrático para sombra. A sobrevivência da Cunhã apresentou efeito quadrático com taxa mínima em 21% de sombreamento. O teor de proteína bruta apresentou efeito de sistema nos quatro ciclos, sendo encontrados maiores teores de proteína bruta no sistema consorciado no 1º e 2º ciclos. Houve ainda efeito quadrático para sombra no 3º ciclo e efeito linear crescente no 4º ciclo. Conclui-se que o sombreamento promove aumento nas taxas de aparecimento e alongamento foliar em níveis intermediários. Além disso, alto nível o sombreamento promove aumento do comprimento foliar, eleva a sobrevivência da Cunhã e diminui a produção de massa de forragem.
Assuntos
Pastagens , Produtos Agrícolas , Conservação de Terras , Poaceae , FabaceaeRESUMO
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.
Assuntos
Feminino , Animais , Ovinos/embriologia , Ovinos/fisiologia , Superovulação , Técnicas de Cultura EmbrionáriaRESUMO
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.
Assuntos
Feminino , Animais , Crioprotetores/química , Ovinos/embriologia , Sacarose/administração & dosagem , VitrificaçãoRESUMO
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.(AU)
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.(AU)
Assuntos
Animais , Feminino , Ovinos/embriologia , Sacarose/administração & dosagem , Crioprotetores/química , VitrificaçãoRESUMO
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.(AU)
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.(AU)
Assuntos
Animais , Feminino , Ovinos/embriologia , Ovinos/fisiologia , Técnicas de Cultura Embrionária , SuperovulaçãoRESUMO
The objective of this study was to evaluate the effectiveness of reusing a controlled internal drug release (CIDR) device for up to three times in the reproductive performance of dairy goats raised in the semi-arid zone of northeastern Brazil. Forty-five goats were allocated into three hormone treatments, as follows: CIDR1x, treated with new CIDR during nine days. Two days prior to device removal, injections of 75 +-g d-cloprostenol and 300 IU equine chorionic gonadotropin (eCG) were administrated. For the other treatments, the same hormone protocol was used, differing only by the use of the same CIDR for a second time in CIDR2x and for a third time in CIDR3x. The interval from device removal to the onset of estrus (13.3 +- 1.1h vs. 13.8 +- 2.6h vs. 13.3 +-1.4h), as well as estrus duration (33.6 +- 7.3h vs. 29.6 +-3.2h vs. 32.8 +- 4.5h), did not differ (p > 0.05) among groups CIDR1x, CIDR2x and CIDR3x, respectively. All synchronized females were found to be in estrus. The overall fertility and prolificacy after artificial insemination were 82.2% and 1.9 kids, respectively, without significant difference (p > 0.05) among treatments. The use of the same CIDR for up to three times was effective using 9-day estrus synchronization protocols in dairy goats.
Objetivou-se avaliar o efeito da utilização do mesmo dispositivo de liberação controlada de drogas (CIDR) por até três vezes sobre o desempenho reprodutivo de cabras leiterias exploradas no semiárido do Nordeste Brasileiro. Foram utilizadas 45 cabras divididas em três tratamentos de sincronização do estro, sendo: CIDR1x, tratadas com CIDR novo durante nove dias. Dois dias antes da retirada do dispositivo, foram aplicados 75 +-g de d-cloprostenol e 300 UI de gonadotrofina coriônica eqüina (eCG). Para os demais tratamentos, foi utilizado o mesmo protocolo hormonal, diferindo apenas pelo uso do mesmo CIDR pela segunda vez no grupo CIDR2x e uso pela terceira vez no grupo CIDR3x. O intervalo entre a retirada do dispositivo e o início do estro (13,3+- 1,1h vs. 13,8 +- 2,6h vs. 13,3 +- 1,4h), bem como, a duração do estro (33,6 +- 7,3h vs. 29,6 +- 3,2h vs. 32,8 +- 4,5h) não diferiram (p > 0,05) entre os grupos CIDR1x, CIDR2x e CIDR3x, respectivamente. Todas as fêmeas sincronizadas foram identificadas em estro. As médias de fertilidade e prolificidade média após inseminação artificial foram, respectivamente, de 82,2% e 1,9 crias, não havendo diferença significativa (p > 0,05) entre os tratamentos. A utilização do mesmo CIDR por até três vezes foi viável na sincronização do estro de caprinos leiteiros.
Assuntos
Animais , Cabras , Fertilidade , Liberação Controlada de Fármacos , HormôniosRESUMO
The objective of this study was to evaluate the effectiveness of reusing a controlled internal drug release (CIDR) device for up to three times in the reproductive performance of dairy goats raised in the semi-arid zone of northeastern Brazil. Forty-five goats were allocated into three hormone treatments, as follows: CIDR1x, treated with new CIDR during nine days. Two days prior to device removal, injections of 75 µg d-cloprostenol and 300 IU equine chorionic gonadotropin (eCG) were administrated. For the other treatments, the same hormone protocol was used, differing only by the use of the same CIDR for a second time in CIDR2x and for a third time in CIDR3x. The interval from device removal to the onset of estrus (13.3 ± 1.1h vs. 13.8 ± 2.6h vs. 13.3 ± 1.4h), as well as estrus duration (33.6 ± 7.3h vs. 29.6 ± 3.2h vs. 32.8 ± 4.5h), did not differ (p > 0.05) among groups CIDR1x, CIDR2x and CIDR3x, respectively. All synchronized females were found to be in estrus. The overall fertility and prolificacy after artificial insemination were 82.2% and 1.9 kids, respectively, without significant difference (p > 0.05) among treatments. The use of the same CIDR for up to three times was effective using 9-day estrus synchronization protocols in dairy goats.
The objective of this study was to evaluate the effectiveness of reusing a controlled internal drug release (CIDR) device for up to three times in the reproductive performance of dairy goats raised in the semi-arid zone of northeastern Brazil. Forty-five goats were allocated into three hormone treatments, as follows: CIDR1x, treated with new CIDR during nine days. Two days prior to device removal, injections of 75 µg d-cloprostenol and 300 IU equine chorionic gonadotropin (eCG) were administrated. For the other treatments, the same hormone protocol was used, differing only by the use of the same CIDR for a second time in CIDR2x and for a third time in CIDR3x. The interval from device removal to the onset of estrus (13.3 ± 1.1h vs. 13.8 ± 2.6h vs. 13.3 ± 1.4h), as well as estrus duration (33.6 ± 7.3h vs. 29.6 ± 3.2h vs. 32.8 ± 4.5h), did not differ (p > 0.05) among groups CIDR1x, CIDR2x and CIDR3x, respectively. All synchronized females were found to be in estrus. The overall fertility and prolificacy after artificial insemination were 82.2% and 1.9 kids, respectively, without significant difference (p > 0.05) among treatments. The use of the same CIDR for up to three times was effective using 9-day estrus synchronization protocols in dairy goats.