RESUMO
BACKGROUND: The proteomic profile of cryopreserved in vitro produced bovine embryos is little known but can provide insights on the successful application of cryo procedures in support of animal breeding. OBJECTIVE: To identify embryonic proteins and biomarkers related to improved cryotolerance of vitrified in vitro produced bovine embryos. MATERIALS AND METHODS: Proteins were isolated from embryo pools (n = 25 embryos per replicate) and analyzed using the nanoLC - MS/MS system. Further, the UniProtKB database (Uniprot -http://www.uniprot.org/) was used for protein identification. Proteins were classified based on their molecular mass, isoelectric point, and enzymatic activity. Post-translational modification predictions and functional gene ontology analysis were performed as well. Finally, a protein-protein interaction network was created to shed light on the embryo interactome. RESULTS: Based on the MS/MS approach, 66 proteins were identified from vitrified Bos taurus embryos. The retrieved proteins were presumably annotated, which allowed a description of the qualitative and functional aspects of the embryo proteome after the vitrification process. CONCLUSION: These findings allowed us to conclude that in vitro-produced vitrified embryos expressed proteins that underlie biological processes related to reproduction, stress and lipid metabolic process, which are essential to maintain embryo viability. doi.org/10.54680/fr22410110512.
Assuntos
Criopreservação , Fertilização in vitro , Bovinos , Animais , Fertilização in vitro/veterinária , Criopreservação/veterinária , Criopreservação/métodos , Espectrometria de Massas em Tandem , Proteômica , Vitrificação , Blastocisto , Técnicas de Cultura Embrionária/veterinária , Técnicas de Cultura Embrionária/métodosRESUMO
BACKGROUND: Semen cryopreservation is essential in animal breeding programs for improving the availability of genetic resources from animals with high breeding value. OBJECTIVE: To evaluate the addition of Brazil nut extract as a replacement for egg yolk in bovine semen cryopreservation. MATERIALS AND METHODS: Semen was collected from five Nelore bulls and cryopreserved with the addition (treatments) of 0, 25, 50, 75, or 100% Brazil nut extract in the cryoprotectant medium. After thawing, spermatic cells were evaluated for morphology, plasma membrane integrity, spermatic kinetics, and in vitro fertilization. The experimental design was in randomized blocks, and the data were submitted to regression analysis. RESULTS: The minor-type and total defects, and plasma membrane integrity were affected (P < 0.05) as a function of egg yolk substitution with Brazil nut extract. There was a significant effect (P < 0.05) of Brazil nut extract addition on the spermatic kinetics and cleavage rate. CONCLUSION: The addition of Brazil nut extract in the cryoprotective medium as a substitute of egg yolk for freezing bovine semen negatively affects sperm quality and fertility.
Assuntos
Bertholletia/química , Criopreservação/veterinária , Crioprotetores , Extratos Vegetais , Preservação do Sêmen , Animais , Bovinos , Crioprotetores/farmacologia , Gema de Ovo , Masculino , Melhoramento Vegetal , Extratos Vegetais/farmacologia , Sêmen , Análise do Sêmen , Preservação do Sêmen/veterinária , Motilidade dos Espermatozoides , EspermatozoidesRESUMO
This study aimed to evaluate the effect of FecGE mutation on the development of ovarian follicles. To this end, 42 Santa Inês ewes were genotyped for FecGE mutation and classified as wild-type (FecG+/+), heterozygous (FecG+/E) or mutant homozygous (FecGE/E). Ovarian fragments were processed, and the follicles were analyzed with regard to the morphology and morphometry using classical histology. For the evaluation of follicular dynamics, ewes underwent oestrous synchronization and were monitored throughout an interovulatory period. A higher (Pâ¯<â¯0.05) percentage of morphologically normal follicles in the primordial stage was identified in FecGE/E (90.0%) and FecG+/E (88.1%) ewes than in the FecG+/+ (73.0%) ewes. There was also a significantly greater (Pâ¯<â¯0.05) number of morphologically normal follicles in the FecGE/E (87.3%) and FecG+/E (83.3%) ewes than in FecG+/+ (76.8%) ewes in the transitional stage. A smaller (Pâ¯<â¯0.05) diameter was observed in the secondary follicles in FecGE/E (93.8⯵m) ewes than in FecG+/E (171.8⯵m) ewes. Regarding follicular dynamics, FecGE/E ewes showed a greater (Pâ¯<â¯0.05) number of ovulations (2.5⯱â¯0.2) than FecG+/+ ewes (1.5⯱â¯0.3) ewes. Ovulatory follicles were smaller (Pâ¯<â¯0.05) in the FecGE/E (5.1â¯mm) and FecG+/E (5.2â¯mm) ewes than in FecG+/+ (5.8â¯mm) ewes. Santa Inês nulliparous ewes carrying the FecGE mutation showed a greater proportion of morphologically normal follicles in the primordial and transitional stages than those not carrying the mutation. FecGE/E ewes demonstrated a higher number of ovulated follicles and that FecGE/E and FecG+/E ewes presented ovulatory follicles with a smaller diameter.
Assuntos
Folículo Ovariano/fisiologia , Ovinos/genética , Ovinos/fisiologia , Animais , Estro/fisiologia , Feminino , Genótipo , Mutação , Ovulação/fisiologia , Ovinos/classificaçãoRESUMO
BACKGROUND: Semen freezing is of great importance for animal production because it allows the use and the rapid diffusion of the genetic material from economically important animals. OBJECTIVE: To evaluate the effect of açai (Euterpe oleracea; Arecaceae family) extract addition to the semen cryopreservation diluent on the morphology, sperm motility parameters, and plasma membrane integrity of spermatozoa. MATERIALS AND METHODS: The ejaculates, obtained from five bulls with low performance on semen freezing, were fractionated and distributed according to the experimental group. The control samples did not have açaí extract added, whereas to the treated groups were added 5, 10, 15 or 20 mg ml-1 of açaí extract into the semen diluent. The sperm morphology was evaluated with a formalin-saline-buffered solution. The plasma membrane integrity was evaluated by the epifluorescent test, while the cellular kinetics was assessed by an automated analysis of the spermatic movement. RESULTS: The sperm defects showed a linearly decreasing effect (P < 0.05) with the addition of different concentrations of açaí extract. The plasma membrane integrity was higher (P < 0.05) after the açaí addition to the cryopreservation diluent. There was no significant effect (P > 0.05) of the açaí extract on the kinetics of spermatozoa. CONCLUSION: The addition of açaí extract to the cryopreservation diluent provided better preservation of the structural integrity of the sperm plasma membrane in the bull's semen with low tolerance to the cryopreservation process.
RESUMO
SummaryPluripotency-associated transcription factors (PATFs) modulate gene expression during early mammalian embryogenesis. Despite a strong understanding of PATFs during mouse embryogenesis, limited progress has been made in ruminants. This work aimed to describe the temporal expression of eight PATFs during both sheep and cattle preimplantation development. Transcript availability of PATFs was evaluated by reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) in eggs, cleavage-stage embryos, morulae, and blastocysts. Transcripts of five genes were detected in all developmental stages of both species (KLF5, OCT4, RONIN, ZFP281, and ZFX). Furthermore, CMYC was detected in all cattle samples but was found from cleavage-stage onwards in sheep. In contrast, NR0B1 was detected in all sheep samples but was not detected in cattle morulae. GLIS1 displayed the most significant variation in temporal expression between species, as this PATF was only detected in cattle eggs and sheep cleavage-stage embryos and blastocysts. In silico analysis suggested that cattle and sheep PATFs share similar size, isometric point and molecular weight. A phenetic analysis showed two patterns of PATF clustering between cattle and sheep, among several mammalian species. In conclusion, the temporal expression of pluripotency-associated transcription factors differs between sheep and cattle, suggesting species-specific regulation during preimplantation development.
Assuntos
Biomarcadores/metabolismo , Blastocisto/metabolismo , Embrião de Mamíferos/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Células-Tronco Pluripotentes/metabolismo , Fatores de Transcrição/metabolismo , Animais , Blastocisto/citologia , Bovinos , Diferenciação Celular , Embrião de Mamíferos/citologia , Feminino , Perfilação da Expressão Gênica , Células-Tronco Pluripotentes/citologia , Ovinos , Fatores de Transcrição/genéticaRESUMO
The aim of this study was to evaluate the pregnancy rate of in vitro produced embryos of Gyr cattle (Bos indicus) after cryopreservation by the vitrification method under field conditions. Blastocysts in different developmental stages were transferred to recipient cows either fresh (n = 140) or warmed after vitrification (n = 138). The pregnancy rates obtained for fresh embryos were 46.15% (initial blastocyst), 46.93% (blastocyst) and 50.0% (expanded blastocyst) at 35 days post-fertilization, and 43.58% (initial blastocyst), 46.93% (blastocyst) and 50.0% (expanded blastocyst) at 60 days. The pregnancy rates after embryo vitrification were 35.0% (initial blastocyst), 42.30% (blastocyst) and 43.47% (expanded blastocyst) at 35 days post-fertilization, and 32.50% (initial blastocyst), 38.46% (blastocyst) and 43.47% (expanded blastocyst) at 60 days. Embryo vitrification or blastocyst developmental stage did not affect pregnancy rates or incidence of embryonic death. In conclusion, vitrification of Gyr (Bos indicus) embryos under field conditions is an efficient method that can be implemented to use surplus in vitro produced embryos without affecting pregnancy rates...
Objetivou-se com o trabalho avaliar a taxa de prenhez a partir de embriões de bovinos da raça Gir (Bos indicus) produzidos in vitro após criopreservação em condições de campo pelo método de vitrificação. Blastocistos em diferentes estádios de desenvolvimento foram transferidos a fresco (n = 140) ou aquecidos após a vitrificação (n = 138). As taxas de prenhez de embriões frescos foram 46,15% (blastocisto inicial), 46,93% (blastocisto) e 50,00% (blastocisto expandido) aos 35 dias e 43,58% (blastocisto inicial), 46,93% (blastocisto) e 50,00% (blastocisto expandido) aos 60 dias pós-fertilização, respectivamente. As taxas de prenhez após a vitrificação foram 35,00% (blastocisto inicial), 42,30% (blastocisto) e 43,47% (blastocisto expandido) aos 35 dias e 32,50% (blastocisto inicial), 38,46% (blastocisto) e 43,47% (blastocisto expandido) aos 60 dias pós-fertilização, respectivamente. A vitrificação do embrião ou estádio do desenvolvimento não afetaram as taxas de prenhez ou incidência de morte embrionária. Em conclusão, a vitrificação de embriões de bovinos da raça Gir (Bos indicus) sob condições de campo é um método eficiente que pode ser implementado para aproveitar os embriões produzidos in vitro excedentes sem afetar a taxa de prenhez...
Assuntos
Animais , Bovinos , Bovinos/classificação , Embrião de Mamíferos , Prenhez , VitrificaçãoRESUMO
The aim of this study was to evaluate the pregnancy rate of in vitro produced embryos of Gyr cattle (Bos indicus) after cryopreservation by the vitrification method under field conditions. Blastocysts in different developmental stages were transferred to recipient cows either fresh (n = 140) or warmed after vitrification (n = 138). The pregnancy rates obtained for fresh embryos were 46.15% (initial blastocyst), 46.93% (blastocyst) and 50.0% (expanded blastocyst) at 35 days post-fertilization, and 43.58% (initial blastocyst), 46.93% (blastocyst) and 50.0% (expanded blastocyst) at 60 days. The pregnancy rates after embryo vitrification were 35.0% (initial blastocyst), 42.30% (blastocyst) and 43.47% (expanded blastocyst) at 35 days post-fertilization, and 32.50% (initial blastocyst), 38.46% (blastocyst) and 43.47% (expanded blastocyst) at 60 days. Embryo vitrification or blastocyst developmental stage did not affect pregnancy rates or incidence of embryonic death. In conclusion, vitrification of Gyr (Bos indicus) embryos under field conditions is an efficient method that can be implemented to use surplus in vitro produced embryos without affecting pregnancy rates...(AU)
Objetivou-se com o trabalho avaliar a taxa de prenhez a partir de embriões de bovinos da raça Gir (Bos indicus) produzidos in vitro após criopreservação em condições de campo pelo método de vitrificação. Blastocistos em diferentes estádios de desenvolvimento foram transferidos a fresco (n = 140) ou aquecidos após a vitrificação (n = 138). As taxas de prenhez de embriões frescos foram 46,15% (blastocisto inicial), 46,93% (blastocisto) e 50,00% (blastocisto expandido) aos 35 dias e 43,58% (blastocisto inicial), 46,93% (blastocisto) e 50,00% (blastocisto expandido) aos 60 dias pós-fertilização, respectivamente. As taxas de prenhez após a vitrificação foram 35,00% (blastocisto inicial), 42,30% (blastocisto) e 43,47% (blastocisto expandido) aos 35 dias e 32,50% (blastocisto inicial), 38,46% (blastocisto) e 43,47% (blastocisto expandido) aos 60 dias pós-fertilização, respectivamente. A vitrificação do embrião ou estádio do desenvolvimento não afetaram as taxas de prenhez ou incidência de morte embrionária. Em conclusão, a vitrificação de embriões de bovinos da raça Gir (Bos indicus) sob condições de campo é um método eficiente que pode ser implementado para aproveitar os embriões produzidos in vitro excedentes sem afetar a taxa de prenhez...(AU)
Assuntos
Animais , Bovinos , Prenhez , Embrião de Mamíferos , Vitrificação , Bovinos/classificaçãoRESUMO
The objective of the present study was to determine if separation distance between bucks and does during two distinct climate seasons could affect the reproductive performance of goats subjected to a 45-day mating season (MS). Anglo Nubian does (n = 120) were kept apart from bucks at distances of 2 m (T1), 300 m (T2), and 2000 m (T3) for 60 days prior to the 45-day MS during two distinct climate seasons [dry season (DS, February to March) and rainy season (RS, September to October)] in Sertânia, Pernambuco state, Brazil. There were no effects of distance of separation between bucks and does in any response variable evaluated. However, during the DS, the mean of the first estrous manifestation varied significantly (P>0.05) between groups [7.13±4.49 (T1), 8.84±5.64 (T2), and 6.37±4.21 (T3) days] and during the RS [7.33±5.74 (T1), 6.60±4.88 (T2) and 8.10±4.87 (T3) days]. Similar (P>0.05) estrous induction rates were found during both the DS [100.00% (T1), 100.00% (T2) and 95.50% (T3)] and the RS [100.00% (T1), 100.00% (T2) and 100.00% (T3)]. The estrous synchronization rate was found to be lower during the DS [36.60%; 30.00% (T1), 35.00% (T2) and 45.00% (T3)] than during the RS [56.60%; 50.00% (T1), 60.00% (T2) and 60.00% (T3)]. Pregnancy rates during the DS [P>0.05; 80.00% (T1), 70.00% (T2) and 75.00% (T3)] were lower than during the RS [P>0.05; 90.00% (T1), 90.00% (T2) and 95.00%(T3)]. In summary, the separation distance between bucks and does did not affect the reproductive outcome of Anglo Nubian goats over a 45-day MS under tropical conditions. Greater reproductive outcome was observed during the RS than the DS regardless of the separation distance between bucks and does...(AU)
O objetivo do presente estudo foi determinar se a distância de isolamento entre machos e fêmeas avaliado durante dois períodos climáticos distintos afeta o desempenho reprodutivo de caprinos submetidos a uma estação de monta (EM) de 45 dias. Fêmeas Anglo Nubianas (n = 120) foram afastadas dos machos por distâncias de 2 m (T1), 300 m (T2) e 2000 m (T3) por 60 dias antes da EM de 45 dias sob diferentes condições climáticas [estação seca (ES, Fevereiro a Março) e estação chuvosa (EC, Setembro a Outubro)] em Sertânia, estado de Pernambuco, Brasil. Não houve efeito da distância de pré-condicionamento entre machos e fêmeas em nenhuma variável avaliada. No entanto, durante a ES, a média da primeira manifestação de estro variou (P>0,05) entre os grupos [7,13±4,49 (T1), 8,84±5,64 (T2) e 6,37±4,21 (T3) dias] e durante a EC [7,33±5,74 (T1), 6,60±4,88 (T2) e 8,10±4,87 (T3)]. A taxa de indução de estro foi semelhante (P>0,05) entre ES [100,00% (T1), 100,00% (T2) e 95,50% (T3)] e EC [100,00% (T1), 100,00% (T2) e 100,00% (T3)]. A sincronização do estro foi inferior durante a ES [36,60%, 30,00% (T1), 35,00% (T2) e 45,00% (T3)] que durante a EC [56,60%, 50,00% (T1), 60,00% (T2) e 60,00% (T3)]. As taxas de prenhez na ES [P>0,05; 80,00% (T1), 70,00% (T2) e 75,00% (T3)] foram menores do que em EC [P>0,05; 90,00% (T1), 90,00% (T2) e 95,00% (T3)]. Em conclusão, a distância de pré-condicionamento entre machos e fêmeas não afetou o desempenho reprodutivo durante a EM de 45 dias de caprinos Anglo Nubianos sob condições tropicais. Um desempenho reprodutivo maior foi observado durante a estação chuvosa comparado a estação seca, independente da distância de pré-condicionamento...(AU)
Assuntos
Animais , Comportamento Sexual Animal , Mudança Climática/estatística & dados numéricos , Ovinos/classificaçãoRESUMO
This study aimed to evaluate the effect of temporary suckling interruption associated with the male effect on reproductive performance of pluriparous Santa Inês ewes. The females were kept apart from the males for 60 days and then randomly distributed into three treatments associated with the male effect (DT0, DT24 and DT48); in DT0, there was no suckling interruption; in DT24, suckling was interrupted for 24 hours, and in DT48, sucking was interrupted for 48 hours. Estrous distribution was observed within 31 (DT0), 27 (DT24) and 38 (DT48) days of the breeding season. Estrous synchronization up to the 5th day of the mating season was observed in 15% (DT0), 30% (DT24) and 25% (DT48) of the females, with no difference among treatments. Estrous percentages were 90% (DT0), 100% (DT24) and 100% (DT48), with no difference among treatments. Pregnancy rates were 38.4% (DT0), 60.0% (DT24) and 45.0% (DT48) with no difference among treatments. Prolificacy was 1.43 (DT0), 1.17 (DT24) and 1.22 (DT48) and did not differ between treatments. In conclusion, temporary suckling interruption associated with the male effect is efficient to induce estrous but not to synchronize estrous or increase the pregnancy rates and prolificacy of Santa Inês ewes during a 45-day breeding season...(AU)
O estudo teve como objetivo avaliar o efeito do desmame temporário associado ao efeito macho sobre o desempenho reprodutivo de ovinos Santa Inês. As fêmeas foram mantidas distantes dos machos por 60 dias e aleatoriamente distribuídas em três tratamentos associados ao efeito macho (DT0, DT24 e DT48), no qual em DT0, não houve interrupção da amamentação; em DT24, amamentação interrompida por 24 horas e em DT48, interrupção da amamentação por 48 horas. Distribuição de estro foi observada em 31 (DT0), 27 (DT24) e 38 (DT48) dias da estação de monta. Sincronização de estro até o quinto dia da estação de monta foi observada em 15% (DT0), 30% (DT24) e 25% (DT48) das fêmeas, não havendo diferença entre os tratamentos. Percentagens de estro foram de 90% (DT0), 100% (DT24) e 100% (DT48), não havendo diferença entre os tratamentos. As taxas de prenhez foram de 38,4% (DT0), 60,0% (DT24) e 45,0% (DT48), sem diferença entre os tratamentos. A prolificidade foi de 1,43 (DT0), 1,17 (DT24) e 1,22 (DT48), e não diferiu entre os tratamentos. Em conclusão, o desmame temporário associado ao efeito macho é eficiente na indução do estro, embora não seja eficiente na sincronização de estros e não aumente as taxas de gestação e prolificidade das ovelhas Santa Inês durante uma estação de monta de 45 dias...(AU)
Assuntos
Animais , Ovinos/classificação , Desmame , Comportamento Sexual Animal , PrenhezRESUMO
The objective of the present study was to determine if separation distance between bucks and does during two distinct climate seasons could affect the reproductive performance of goats subjected to a 45-day mating season (MS). Anglo Nubian does (n = 120) were kept apart from bucks at distances of 2 m (T1), 300 m (T2), and 2000 m (T3) for 60 days prior to the 45-day MS during two distinct climate seasons [dry season (DS, February to March) and rainy season (RS, September to October)] in Sertânia, Pernambuco state, Brazil. There were no effects of distance of separation between bucks and does in any response variable evaluated. However, during the DS, the mean of the first estrous manifestation varied significantly (P>0.05) between groups [7.13±4.49 (T1), 8.84±5.64 (T2), and 6.37±4.21 (T3) days] and during the RS [7.33±5.74 (T1), 6.60±4.88 (T2) and 8.10±4.87 (T3) days]. Similar (P>0.05) estrous induction rates were found during both the DS [100.00% (T1), 100.00% (T2) and 95.50% (T3)] and the RS [100.00% (T1), 100.00% (T2) and 100.00% (T3)]. The estrous synchronization rate was found to be lower during the DS [36.60%; 30.00% (T1), 35.00% (T2) and 45.00% (T3)] than during the RS [56.60%; 50.00% (T1), 60.00% (T2) and 60.00% (T3)]. Pregnancy rates during the DS [P>0.05; 80.00% (T1), 70.00% (T2) and 75.00% (T3)] were lower than during the RS [P>0.05; 90.00% (T1), 90.00% (T2) and 95.00%(T3)]. In summary, the separation distance between bucks and does did not affect the reproductive outcome of Anglo Nubian goats over a 45-day MS under tropical conditions. Greater reproductive outcome was observed during the RS than the DS regardless of the separation distance between bucks and does...
O objetivo do presente estudo foi determinar se a distância de isolamento entre machos e fêmeas avaliado durante dois períodos climáticos distintos afeta o desempenho reprodutivo de caprinos submetidos a uma estação de monta (EM) de 45 dias. Fêmeas Anglo Nubianas (n = 120) foram afastadas dos machos por distâncias de 2 m (T1), 300 m (T2) e 2000 m (T3) por 60 dias antes da EM de 45 dias sob diferentes condições climáticas [estação seca (ES, Fevereiro a Março) e estação chuvosa (EC, Setembro a Outubro)] em Sertânia, estado de Pernambuco, Brasil. Não houve efeito da distância de pré-condicionamento entre machos e fêmeas em nenhuma variável avaliada. No entanto, durante a ES, a média da primeira manifestação de estro variou (P>0,05) entre os grupos [7,13±4,49 (T1), 8,84±5,64 (T2) e 6,37±4,21 (T3) dias] e durante a EC [7,33±5,74 (T1), 6,60±4,88 (T2) e 8,10±4,87 (T3)]. A taxa de indução de estro foi semelhante (P>0,05) entre ES [100,00% (T1), 100,00% (T2) e 95,50% (T3)] e EC [100,00% (T1), 100,00% (T2) e 100,00% (T3)]. A sincronização do estro foi inferior durante a ES [36,60%, 30,00% (T1), 35,00% (T2) e 45,00% (T3)] que durante a EC [56,60%, 50,00% (T1), 60,00% (T2) e 60,00% (T3)]. As taxas de prenhez na ES [P>0,05; 80,00% (T1), 70,00% (T2) e 75,00% (T3)] foram menores do que em EC [P>0,05; 90,00% (T1), 90,00% (T2) e 95,00% (T3)]. Em conclusão, a distância de pré-condicionamento entre machos e fêmeas não afetou o desempenho reprodutivo durante a EM de 45 dias de caprinos Anglo Nubianos sob condições tropicais. Um desempenho reprodutivo maior foi observado durante a estação chuvosa comparado a estação seca, independente da distância de pré-condicionamento...
Assuntos
Animais , Comportamento Sexual Animal , Mudança Climática/estatística & dados numéricos , Ovinos/classificaçãoRESUMO
This study aimed to evaluate the effect of temporary suckling interruption associated with the male effect on reproductive performance of pluriparous Santa Inês ewes. The females were kept apart from the males for 60 days and then randomly distributed into three treatments associated with the male effect (DT0, DT24 and DT48); in DT0, there was no suckling interruption; in DT24, suckling was interrupted for 24 hours, and in DT48, sucking was interrupted for 48 hours. Estrous distribution was observed within 31 (DT0), 27 (DT24) and 38 (DT48) days of the breeding season. Estrous synchronization up to the 5th day of the mating season was observed in 15% (DT0), 30% (DT24) and 25% (DT48) of the females, with no difference among treatments. Estrous percentages were 90% (DT0), 100% (DT24) and 100% (DT48), with no difference among treatments. Pregnancy rates were 38.4% (DT0), 60.0% (DT24) and 45.0% (DT48) with no difference among treatments. Prolificacy was 1.43 (DT0), 1.17 (DT24) and 1.22 (DT48) and did not differ between treatments. In conclusion, temporary suckling interruption associated with the male effect is efficient to induce estrous but not to synchronize estrous or increase the pregnancy rates and prolificacy of Santa Inês ewes during a 45-day breeding season...
O estudo teve como objetivo avaliar o efeito do desmame temporário associado ao efeito macho sobre o desempenho reprodutivo de ovinos Santa Inês. As fêmeas foram mantidas distantes dos machos por 60 dias e aleatoriamente distribuídas em três tratamentos associados ao efeito macho (DT0, DT24 e DT48), no qual em DT0, não houve interrupção da amamentação; em DT24, amamentação interrompida por 24 horas e em DT48, interrupção da amamentação por 48 horas. Distribuição de estro foi observada em 31 (DT0), 27 (DT24) e 38 (DT48) dias da estação de monta. Sincronização de estro até o quinto dia da estação de monta foi observada em 15% (DT0), 30% (DT24) e 25% (DT48) das fêmeas, não havendo diferença entre os tratamentos. Percentagens de estro foram de 90% (DT0), 100% (DT24) e 100% (DT48), não havendo diferença entre os tratamentos. As taxas de prenhez foram de 38,4% (DT0), 60,0% (DT24) e 45,0% (DT48), sem diferença entre os tratamentos. A prolificidade foi de 1,43 (DT0), 1,17 (DT24) e 1,22 (DT48), e não diferiu entre os tratamentos. Em conclusão, o desmame temporário associado ao efeito macho é eficiente na indução do estro, embora não seja eficiente na sincronização de estros e não aumente as taxas de gestação e prolificidade das ovelhas Santa Inês durante uma estação de monta de 45 dias...
Assuntos
Animais , Comportamento Sexual Animal , Desmame , Ovinos/classificação , PrenhezRESUMO
The objective was to evaluate the effects of insulin-like growth factor-I (IGF-I) on the quality and fertility of frozen/thawed ovine semen. Five rams (five ejaculates/ram) were used for evaluation of semen parameters. Before cryopreservation, ejaculates were divided into four aliquots and extended with Tris alone or supplemented with human IGF-I (50, 100, or 250 ng/mL). Semen was evaluated immediately after thawing (T0), after 1 h (T1) and 2 h (T2) post-incubation at 37 °C. The percentage of live cells (fluorescence analysis-calcein and ethidium), acrosome integrity (NAR) and motility were analyzed, and hypo-osmotic swelling tests (HOST) were used to evaluate membrane resistance. In addition, AI was performed using 121 ewes to compare the optimal concentration of IGF-I vs. Tris alone on pregnancy rates after laparoscopic insemination. Pregnancy diagnosis was performed by transrectal ultrasonography. After 1 and 2 h post-incubation, in every group, percentage motile sperm, NAR and HOST decreased compared to semen at T0. Motility was higher (P < 0.05) in the IGF-I 100 and IGF-I 250 groups when compared to the IGF-I 50 and Tris groups (76.2 and 74.4% vs. 66.2 and 64.4 percent, respectively) at T0, after 1 h (67 and 63.6% vs. 56.2 and 54.7%) and 2 h post-incubation (58.2 and 55.8% vs. 48 and 47.2%). Furthermore, viability was higher (P < 0.05) in the insulin-like growth factor-I (IGF-I) 100 and IGF-I 250 groups than in the IGF-I 50 and Tris groups (88.7 and 88.3% vs. 76.6 and 77.6%, respectively) at T0. There was no difference (P > 0.05) in NAR or hypo-osmotic swelling tests (HOST) among groups. There were no differences (P > 0.05) in fertility between the IGF-I 100 and Tris groups. In conclusion, IGF-I improved subjective sperm motility and structural integrity of the plasma membrane without a significant effect on 45-day pregnancy rates after laparoscopic insemination of ewes with frozen-thawed semen.
Assuntos
Criopreservação , Fertilidade/efeitos dos fármacos , Fator de Crescimento Insulin-Like I/farmacologia , Preservação do Sêmen , Sêmen/efeitos dos fármacos , Ovinos , Acrossomo/efeitos dos fármacos , Acrossomo/fisiologia , Animais , Sobrevivência Celular/efeitos dos fármacos , Criopreservação/veterinária , Avaliação Pré-Clínica de Medicamentos , Feminino , Fertilidade/fisiologia , Inseminação Artificial/veterinária , Masculino , Pressão Osmótica/efeitos dos fármacos , Pressão Osmótica/fisiologia , Gravidez , Taxa de Gravidez , Sêmen/citologia , Sêmen/fisiologia , Análise do Sêmen/veterinária , Preservação do Sêmen/efeitos adversos , Preservação do Sêmen/métodos , Preservação do Sêmen/veterinária , Ovinos/fisiologiaRESUMO
The aim of the present study was to identify the migration period of the genital tubercle and its later differentiation into external genital structures in fetuses derived from natural mating and fetuses from fresh, frozen and vitrified embryo transfer. A transrectal ultrasound with a double-frequency linear transducer (6.0 and 8.0 MHz) was used to monitor 123 goat fetuses, which were allocated to one of four groups: fetuses originating from controlled natural mating (G1, n = 32) and fetuses derived from fresh (G2, n = 34), frozen (G3, n = 30) and vitrified (G4, n = 27) embryo transfer. The transferable embryos were collected 7 days after mating by laparoscopy. Migration of the genital tubercle occurred significantly earlier (P < 0.05) in G1 than in G2, G3 and G4. The visualisation of the scrotum, prepuce and vulva occurred significantly earlier (P < 0.05) in G1 than in G2, G3 and G4. Our results show that fetal sexing is feasible after 55 days for fetuses from natural mating and after 60 days in fetuses from fresh and cryopreserved embryos. Thus, real-time ultrasonography is a reliable tool for fetal sex determination in goats after Day 50 of pregnancy, taking into account both the location of the genital tubercle and the identification of external genital structures.
Assuntos
Cabras/embriologia , Análise para Determinação do Sexo/veterinária , Ultrassonografia Pré-Natal/veterinária , Animais , Criopreservação/veterinária , Transferência Embrionária/métodos , Transferência Embrionária/veterinária , Feminino , Idade Gestacional , Masculino , Gravidez , Reprodutibilidade dos Testes , Análise para Determinação do Sexo/métodos , Ultrassonografia Pré-Natal/métodosRESUMO
The objective of this study was to evaluate the effect of retinol (RT) and retinoic acid (RA) on the in vitro development of pre-implantation goat embryos cultured in potassium simplex optimized medium or synthetic oviduct fluid or cocultured in oviductal cells monolayer either in potassium simplex optimized medium or synthetic oviduct fluid. A total of 2407 cumulus-oocyte complexes were aspirated from 2 to 6 mm ovarian follicles from slaughtered animals. Selected cumulus-oocyte complexes were subjected to in vitro maturation in TCM 199 for 24 h at 39 °C in an atmosphere of 5% (v/v) CO(2) in humidified air. In vitro fertilization was performed in modified defined medium. Eighteen hours after in vitro fertilization, cumulus cells were removed and presumptive zygotes were randomly distributed into experimental groups. In Experiment 1, presumptive zygotes were cultured in potassium simplex optimized medium, potassium simplex optimized medium + RT, potassium simplex optimized medium + retinoic acid, synthetic oviduct fluid, synthetic oviduct fluid + RT and synthetic oviduct fluid + RA at 39 °C in a humidified atmosphere of 5% (v/v) CO(2), 5% (v/v) O(2) and 90% (v/v) N(2). In Experiment 2, presumptive zygotes were cocultured in potassium simplex optimized medium + oviductal cells monolayer, potassium simplex optimized medium + RT + oviductal cells monolayer, potassium simplex optimized medium + RA + oviductal cells monolayer, synthetic oviduct fluid + oviductal cells monolayer, synthetic oviduct fluid + RT + oviductal cells monolayer and synthetic oviduct fluid + RA + oviductal cells monolayer in an atmosphere of 5% (v/v) CO(2) in humidified air. In both experiments, media were partially changed on day 2 after in vitro fertilization and unfertilized oocytes were excluded from the experiment. Embryos were cultured or cocultured for 8 days. In Experiment 1, there was no effect of RT or RA supplementation on the proportion of oocytes that reached the morula or blastocyst stages. By contrast, Experiment 2 demonstrated that the addition of 0.28 µg/ml RT and 0.5 µm RA to the embryo culture media stimulated (p < 0.05) development to the morula and blastocyst stages under the coculture conditions tested. In conclusion, retinoids play an important role in pre-implantation development of goat embryos and can be used to enhance in vitro embryo production.
Assuntos
Blastocisto/efeitos dos fármacos , Blastocisto/fisiologia , Técnicas de Cultura Embrionária/veterinária , Cabras/fisiologia , Tretinoína/farmacologia , Vitamina A/farmacologia , Animais , Meios de Cultura , Técnicas de Cultura Embrionária/métodos , Desenvolvimento Embrionário/efeitos dos fármacosRESUMO
The effects of α-tocopherol and ternatin on the morphology, activation, and growth of goat preantral follicles in vitro cultured, for one or five days, were evaluated. Ovarian fragments were immediately fixed (non-cultured control) or in vitro cultured for one or five days in Minimum Essential Medium (MEM) with or without α-tocopherol or ternatin supplementation, both at concentrations of 5, 10, or 15µM, corresponding to the following treatments: MEM, TOC5, TOC10, TOC 15, TER5, TER10, and TER15. The percentages of morphologically normal preantral follicles in non-cultured ovarian tissue (control) was 73.2 percent and after five days of culture, there was a decrease on these percentages in all treatments (P<0.05) when compared with non-cultured control. Culture of ovarian cortex for five days increased the percentages of follicular activation in all treatments (P<0.05). Ultrastructural analysis did not confirm the integrity of caprine preantral follicles cultured for five days in medium containing antioxidants. This study demonstrated that α-tocopherol and ternatin can promote follicular activation; however, addition of these antioxidants in the tested concentrations reduced the follicular viability after in vitro culture.
Os efeitos do α-tocoferol e da ternatina sobre morfologia, ativação e crescimento de folículos pré-antrais caprinos cultivados in vitro, por um ou cinco dias, foram avaliados. Os fragmentos ovarianos foram imediatamente fixados (controle não-cultivado) ou cultivados in vitro, por um ou cinco dias, em Meio Essencial Mínimo (MEM) com ou sem suplementação com α-tocoferol ou ternatina nas concentrações de 5, 10 ou 15µM, formando os tratamentos MEM, TOC5, TOC10, TOC 15, TER5, TER10, TER15. O percentual de folículos pré-antrais normais no controle não-cultivado foi de 73,2 por cento, depois de cinco dias de cultivo, houve redução desse percentual em todos os tratamentos, quando comparados com o controle não-cultivado (P<0,05). O cultivo por cinco dias aumentou a ativação folicular em todos os tratamentos (P<0,05). A análise ultra-estrutural não mostrou folículos pré-antrais íntegros após cinco dias de cultivo em meio contendo antioxidantes. Concluiu-se que o α-tocoferol e a ternatina podem promover a ativação folicular, no entanto a adição desses antioxidantes nas concentrações testadas reduziu a viabilidade folicular após o cultivo in vitro.
Assuntos
Animais , Antioxidantes/farmacologia , Folículo Ovariano , alfa-Tocoferol/farmacologia , Folículo Ovariano , CabrasRESUMO
The effects of α-tocopherol and ternatin on the morphology, activation, and growth of goat preantral follicles in vitro cultured, for one or five days, were evaluated. Ovarian fragments were immediately fixed (non-cultured control) or in vitro cultured for one or five days in Minimum Essential Medium (MEM) with or without α-tocopherol or ternatin supplementation, both at concentrations of 5, 10, or 15µM, corresponding to the following treatments: MEM, TOC5, TOC10, TOC 15, TER5, TER10, and TER15. The percentages of morphologically normal preantral follicles in non-cultured ovarian tissue (control) was 73.2 percent and after five days of culture, there was a decrease on these percentages in all treatments (P<0.05) when compared with non-cultured control. Culture of ovarian cortex for five days increased the percentages of follicular activation in all treatments (P<0.05). Ultrastructural analysis did not confirm the integrity of caprine preantral follicles cultured for five days in medium containing antioxidants. This study demonstrated that α-tocopherol and ternatin can promote follicular activation; however, addition of these antioxidants in the tested concentrations reduced the follicular viability after in vitro culture.(AU)
Os efeitos do α-tocoferol e da ternatina sobre morfologia, ativação e crescimento de folículos pré-antrais caprinos cultivados in vitro, por um ou cinco dias, foram avaliados. Os fragmentos ovarianos foram imediatamente fixados (controle não-cultivado) ou cultivados in vitro, por um ou cinco dias, em Meio Essencial Mínimo (MEM) com ou sem suplementação com α-tocoferol ou ternatina nas concentrações de 5, 10 ou 15µM, formando os tratamentos MEM, TOC5, TOC10, TOC 15, TER5, TER10, TER15. O percentual de folículos pré-antrais normais no controle não-cultivado foi de 73,2 por cento, depois de cinco dias de cultivo, houve redução desse percentual em todos os tratamentos, quando comparados com o controle não-cultivado (P<0,05). O cultivo por cinco dias aumentou a ativação folicular em todos os tratamentos (P<0,05). A análise ultra-estrutural não mostrou folículos pré-antrais íntegros após cinco dias de cultivo em meio contendo antioxidantes. Concluiu-se que o α-tocoferol e a ternatina podem promover a ativação folicular, no entanto a adição desses antioxidantes nas concentrações testadas reduziu a viabilidade folicular após o cultivo in vitro.(AU)
Assuntos
Animais , alfa-Tocoferol/efeitos adversos , Antioxidantes/efeitos adversos , Folículo Ovariano , CabrasRESUMO
The aim of this study was to determine the period of genital tubercle (GT) migration using ultrasonography in Morada Nova sheep foetuses (n = 117) from natural mating (NM) and frozen embryo transfer (ET) to determine the window when foetal sexing can be determined. The examinations were performed using transrectal ultrasonography with a dual-frequency linear transducer (6.0 and 8.0 MHz) from day 30-54 of pregnancy at 48-h intervals. The period of GT migration of foetuses produced by NM varied from 36 to 46 days of pregnancy, resulting in an average of 39.5 +/- 2.9 days. For foetuses derived from ET, GT migration varied from 42 to 52 days of pregnancy with an average of 48.5 +/- 3.3 days, being possible the GT of foetuses from ET start to migrate 96 h later even if they are of the same gender. Migration of the GT occurred earlier (p < 0.05) in foetuses produced by NM and sexing accuracy for triplet pregnancies (77.8%) was significantly inferior (p < 0.05) to single (100%) and twin (92.9%) pregnancies for foetuses derived by NM. The results allow one to conclude that foetal sexing can be done from the 50th day onwards in foetuses produced by NM and from the 55th day onwards in foetuses derived from ET, and that multiple pregnancies compromise the sexing accuracy by ultrasonography.
Assuntos
Análise para Determinação do Sexo/veterinária , Ovinos/embriologia , Ultrassonografia Pré-Natal/veterinária , Animais , Transferência Embrionária/veterinária , Feminino , Tamanho da Ninhada de Vivíparos , Gravidez , Análise para Determinação do Sexo/métodos , Fatores de Tempo , Ultrassonografia Pré-Natal/métodosRESUMO
In order to improve fetal sexing in the Dorper sheep breed, the objective of the present study was to determine, by repeated ultrasonographic examinations, the migration period of the genital tubercle (GT) in sheep fetuses derived from natural mating or embryo transfer and to compare the accuracy of a single examination with repeated examinations at short intervals. For this purpose, transrectal ultrasound was performed, using a double-frequency linear transducer (6.0 and 8.0 MHz) for monitoring 51 sheep fetuses distributed in three experimental groups (EI, EII and EIII). The fetuses in EI (n = 23) and EII (n = 18) derived, respectively, from natural mating and embryo transfer were monitored at 48-h intervals from the 30th to 60th day of gestation and sexed based on the final location of the GT. The fetuses in EIII (n = 10), which originated from embryo transfer, were examined only once on the 65th day of gestation and sexed taking into consideration the final position of the GT and/or by identification of anatomical structures of external genitalia. The accuracy of fetal sexing was 91.3% (21 fetuses sexed/23 quantified) in EI, 88.9% (16 sexed/18 quantified) in EII and 100% (10 sexed/10 quantified) in EIII, without significant difference (P > 0.05) between experiments. Migration of the GT occurred earlier (P < 0.05) in fetuses produced by natural mating (43.0 +/- 2.8 days) than in those derived from embryo transfer (46.1 +/- 4.7 days). The results show that fetal sexing can be done from the 50th day onward in fetuses produced by natural mating and from the 60th day onward in fetuses derived from frozen embryos. It can also be concluded that repeated ultrasonographic exams in short time intervals do not maximise the accuracy of fetal sexing. In addition, real-time ultrasonography is a reliable tool for fetal sex determination in sheep after Day 50 of gestation, taking into account both the location of the GT and the identification of external genital structures.
Assuntos
Cruzamento , Transferência Embrionária , Análise para Determinação do Sexo/métodos , Carneiro Doméstico/embriologia , Ultrassonografia Pré-Natal , Animais , Feminino , Desenvolvimento FetalRESUMO
Experiments were conducted to investigate the beneficial effects of adding retinol (RT) and retinoic acid (RA) to bovine oocyte maturation media and insulin-like growth factor-I (IGF-I) to embryo culture under chemically-defined conditions. In Experiment 1.1, in vitro maturation (IVM) was performed in basic maturation media (bMM) and supplemented with 0.3microM RT or 0.5microM RA. For embryo development presumptive zygotes and embryos were placed in droplets of potassium simplex optimized medium (KSOM). Addition of RT and RA to bMM improved (p<0.05) blastocyst formation as compared with control treatments. In Experiment 1.2, using embryos originating from oocytes previously treated with RT and RA, the presumptive zygotes were placed in droplets of KSOM and embryos (2-4 cells) in droplets of fresh KSOM supplemented or not with IGF-I. The number of 2-4-cell stage embryos developing to the blastocyst and expanded blastocyst stages were greater (p<0.05) when embryo culture media was supplemented with IGF-I. In Experiment 2.1, IVM was conducted with bMM+FSH containing 0.3microM RT or 0.5microM RA. For embryo development, presumptive zygotes were placed in droplets of KSOM. Addition of RT or RA to IVM medium also enhanced (p<0.05) blastocyst formation. The supplementation of embryo culture media with IGF-I resulted in a greater number (p<0.05) of 2-4-cell stage embryos developing into blastocysts, expanded blastocysts and hatched blastocysts. In Experiment 2.2, using embryos originating from oocytes previously treated with RT and RA, presumptive zygotes were also placed in droplets of KSOM and embryos (2-4 cells) in droplets of fresh KSOM supplemented or not with IGF-I. The supplementation of embryo culture media with IGF-I resulted in a greater (p<0.05) number of 2-4-cell stage embryos developing to the blastocyst, expanded blastocyst and hatched blastocyst stages.
Assuntos
Bovinos/embriologia , Desenvolvimento Embrionário/efeitos dos fármacos , Fator de Crescimento Insulin-Like I/farmacologia , Retinoides/farmacologia , Animais , Blastocisto/efeitos dos fármacos , Blastocisto/fisiologia , Técnicas de Cultura Embrionária/veterinária , Feminino , Fertilização in vitro/veterinária , Masculino , Tretinoína/farmacologia , Vitamina A/farmacologiaRESUMO
The objective of this study was to evaluate the effect of retinol on the in vitro development of early embryos of cultured Bos indicus (Expt 1) to the blastocyst stage in medium simplex of optimization (KSOM) or sintetic fluid of oviduct (SOF) or co-cultured (Expt 2) with an oviduct cell monolayer (OCM) in KSOM or SOF. A total of 3149 cumulus-oocyte complexes obtained by aspirating follicles (2-5 mm diameter) from ovaries of slaughtered animals were selected for IVM and incubated in TCM 199 supplemented with 25 mM HEPES at 39 degrees C in air with 5% CO(2) and maximum humidity for 24 h. In vitro fertilization (IVF) was performed in modified defined medium (mDM) medium. Eighteen hours after IVF, cumulus cells were removed and presumptive zygotes were randomly allocated to the experimental groups. Zygotes cultured (Expt 1) in KSOM + retinol, KSOM, SOF + retinol and SOF were incubated in maximum humidity at 39 degrees C, 5% CO(2), 5% O(2) and 90% N(2). Zygotes co-cultured (Expt 2) in KSOM + retinol + OCM, KSOM + OCM, SOF + retinol + OCM and SOF + OCM were incubated at 39 degrees C, 5% CO(2). In both experiments media were partially changed 48 h after IVF and unfertilized ova were removed. Afterwards embryos were kept in culture or co-culture for further 9 days. In Expt 1, blastocyst rates (day 7) were 14.6% (KSOM + retinol), 15.8% (KSOM), 16.4% (SOF + retinol) and 15.9% (SOF). In Expt 2, the blastocyst rates (day 7) were 25.4% (KSOM + retinol + OCM) 14.2% (KSOM + OCM), 24.3% (SOF + retinol + OCM) and 15.9% (SOF + OCM). The same influence profile of retinol was observed in the formation of the expanded (day 9) and hatched (day 11) blastocysts. The results obtained in Expt 2 demonstrated that the addition of 0.28 microg/ml retinol to the embryo culture media used in this study had a significant (p < 0.05) positive effect on bovine early embryonic development, under the conditions tested, and can be used to enhance in vitro embryo production.