Exogenous paraoxonase-1 during oocyte maturation improves bovine embryo development in vitro.

The aims of this study were (i) to measure the direct effects of exogenous human recombinant PON1 (rPON1) on bovine oocyte maturation at the molecular level (gene expression) and (ii) to measure the carry-over effects of PON1 on pre-implantation embryo development in vitro.

Eficiência da recuperação de embriões e os efeitos de consecutivas colheitas sobre o aparelho reprodutor de doadoras da espécie caprina.

Resumo: 0 objetivo deste experimento foi comparar a eficiêncÎa e o efeito de consecutivas colheitas de embriões, por três diferentes métodos (transcervical-T1, laparoscopia-T2 e laparotomia-T3), sobre a atividade reprodutiva de doadoras da espécie caprina. Utilizaram-se 10 cabras em cada método (T1, T2 e T3), sendo as colheitas de embriões repetidas três vezes consecutivas nas mesmas fêmeas, com intervalo de 56 dias. As fêmeas foram sincronizadas com esponjas vaginais impregnadas com 60 mg de acetato de medroxiprogesterona durante 10 dias e 100 ug de cloprostenol aplicados pela via lM no oitavo dia da sincronizaçao. No 8º dia, iniciou-se a superovulaçao com 250 UI de FSH de origem suína, divididas em oito doses decrescentes, aplicadas em intervalo de 12 horas. As fêmeas foram acasaladas e as colheitas de embriões realizadas no 5° ou 6° dia após a ultima cobertura. Após 56 dias da terceira colheita de embriões, foram realizados o abate e a necrópsia das doadoras. 0 tempo necessário para a a colheita de embriões em cada método foi de 21 minutos e 32 segundos; 37 minutos e 14 segundos e 56 minutos e 22 segundos, respectivamente, para TI, T 2 e T 3 (p<0,0I). A maior taxa de recuperaçao da soluçao de lavagem foi no T3 (83,7%), seguido por T2 (72,2%) e TI (64,3%) (p<0,05). As taxas médias de recuperaçao dos embriões foram 57,1; 81,1 e 27,3% para TI' T2 e T3' respectivamente, com variaçao entre 0-100%. A taxa de recuperaçao de embriões sofreu influência de varios fatores, com a presença de corpos luteos regredidos, a taxa de ovulaçao e a presença de aderências no sistema genital, mas, isoladamente, a taxa de recuperaçao de embriões foi satisfatória nos três métodos. 0 T1 causou eversao do endométrio e aderência entre o corno uterino e o epíploo em uma unica fêmea, o T2 causou eversao do endométrio em 30%, 40% e 60% e aderências do sistema genital em 10%, 10% e 70% das fêmeas à 1 a, 2a e 3a colheitas, respectivamente. 0 T3 causou aderências no sistema genital em 80% das doadoras ; após a primeira e 100% após a segunda colheita. 0 T1 e o T2 permitem o usa de doadoras em repetidas colheitas de embriões, o que nao ocorre com o T3, que causa aderências no sistema genital e órgaos circunvizinhos em 100% dos casos. [Efficiency and effect of consecutive embryo recoveries on the reproductive system of goat donnors].

Sincronização de estro com CIDR reutilizado em cabras lactantes da raça Toggenburg tratadas com somatotropina bovina recombinante (r-bST).

Avaliou-se a resposta de cabras tratadas com r-bST no protocolo de sincronização de estro. Foram utilizadas 26 cabras Toggenburg, divididas em dois tratamentos: T1 (n=13), tratadas com quatro injeções de ..250mg de rbST, a intervalos de 14 dias, e T2 (n=13), tratadas com solução salina (controle). Na semana seguinte à última injeção da r-bST, colocou-se o dispositivo intravaginal com progesterona (dia 0), previamente utilizado por cinco dias, e injetou-se PGF2.. (22,5µg) nos animais dos dois tratamentos, e o dispositivo foi retirado no dia 6. Todas as fêmeas em estro foram submetidas à monta natural. A porcentagem de animais em estro e a taxa de gestação foram 76,9 e 70,0 e 84,6 e 72,7%, no T1 e T2, respectivamente. Não houve diferença (P>0,05) na duração do estro, no intervalo tratamento-início do estro, no número de ovulações, nos intervalos: início e final do estro à ovulação e retirada do dispositivo à ovulação entre os animais dos dois tratamentos. O diâmetro médio dos folículos ovulatórios das fêmeas não diferiu (P>0,05). Durante a permanência do dispositivo, as concentrações séricas de progesterona apresentaram valores semelhantes (P>0,05) entre as cabras de T1 e T2. A r-bST não afetou a sincronização de estro.

Isolation and characterization of mesenchymal stem cells derived from bovine Wharton's jelly and their potential for use in cloning by nuclear transfer.

RESUMO: A geleia de Wharton é uma fonte de células tronco mesenquimais (CTMs) que ainda não havia sido testada para a produção de embriões bovinos por transferência nuclear (TN). O objetivo deste estudo foi isolar, caracterizar e testar as CTMs derivadas da geleia de Wharton para produção de embriões e gestações por transferência nuclear em bovinos. O cordão umbilical foi coletado durante o nascimento e as células derivadas da geleia de Wharton (CGWs) foram isoladas por explante e cultivadas em Dulbecco?s Modified Eagle Medium. Fibroblastos (FB) da pele foram isolados após 6 meses de vida. As análises morfológicas foram realizadas pelas microscopias de campo claro e eletrônica de varredura durante o cultivo celular. Caracterização fenotípica e genotípica por citometria de fluxo, imunocitoquímica, RT-PCR e indução da diferenciação em linhagens celulares foi realizada com as CGWs. No procedimento de TN, ovócitos no estágio de metáfase II foram enucleados usando micromanipuladores, fusionados com CGWs ou FB e então ativados artificialmente. Micrografias de microscopia de varredura revelaram que CGWs tiveram forma variada sob cultivo. Os marcadores mesenquimais de CTMs (CD29+, CD73+, CD90+ and CD105+) foram expressos em cultura de CGWs bovina, como evidenciado por citometria de fluxo, imunocitoquímica e RT-PCR. Quando induzidas, estas células diferenciaram-se em osteócitos, condrócitos e adipócitos. Após classificação, as CGWs foram utilizadas na TN. A taxa de formação de blastocistos por TN com CGWs no sétimo dia de cultivo foi de 25,80±0,03%, similar a produção de blastócitos por TN com fibroblastos de pele (19,00±0,07). Gestações foram obtidas e mostraram que CGWs constituem um novo tipo celular para ser usado na clonagem animal. ABSTRACT: Wharton?s jelly is a source of mesenchymal stem cells (MSCs) that had not yet been tested for bovine embryo production by nuclear transfer (NT). Thus, the objective of this study was to isolate, characterize and test MSCs derived from Wharton?s jelly for embryo and pregnancy production by NT in cattle. The umbilical cord was collected during calving and cells derived from Wharton?s jelly (WJCs) were isolated by explant and cultured in Dulbecco?s Modified Eagle Medium. Skin Fibroblasts (FB) were isolated after 6 months of life. Morphological analysis was performed by bright field and scanning electron microscopy (SEM) during cell culture. Phenotypic and genotypic characterization by flow cytometry, immunocytochemistry, RT-PCR and differentiation induction in cell lineages were performed for WJC. In the NT procedure, oocytes at the arrested metaphase II stage were enucleated using micromanipulators, fused with WJCs or FB and later activated artificially. SEM micrographs revealed that WJCs have variable shape under culture. Mesenchymal markers of MSCs (CD29+, CD73+, CD90+ and CD105+) were expressed in bovine-derived WJC cultures, as evidenced by flow cytometry, immunocytochemistry and RT-PCR. When induced, these cells differentiated into osteocytes, chondrocytes and adipocytes. After classification, the WJCs were used in NT. Blastocyst formation rate by NT with WJCs at day 7 was 25.80±0.03%, similar to blatocyst rate with NT using skin fibroblasts (19.00±0.07%). Pregnancies were obtained and showed that WJCs constitute a new cell type for use in animal cloning.

Doppler ultrasound in the diagnosis of early pregnancy in sheep.

2016

Color Doppler ultrasound as a substitute to laparoscopy for the CL count in superovulated sheep.

This study aimed to evaluate the Color Doppler ultrasound as a substitute for laparoscopy for couting of corpora lutea (CL) in superovulated sheep. In conclusion, the Color Doppler ultrasonography is highly efficient to estimate the number of CLs in superovulated ewes. This represents an important advance because it replaces invasive laparoscopic procedure, avoids fasting, drugs use and unnecessary handling in animals that did not respond to the treatment. Therefore, the Color Doppler ultrasound can replace the laparoscopy for the assessment of superovulated sheep.

Correlations between testicular surface temperatures measured by infrared thermography and seminal parameters in rams of distinct genotypic groups.

The aim of this study was to correlate the testicular surface temperature with rectal temperature and semen quality in rams of different genotypic groups.

Effect of the FSH dose in superovulatory protocols on ovulatory follicle dynamics in ewes.

The objective of this study, was to evaluate the exogenous FSH dose effect on gonadotrophic treatment over ewes ovulatory follicle dynamics.

Follicular population on the estimation of the superovulatory response in Santa Ines sheep.

The aim was to verify the correlation between follicular population count, superovulatory response and the recovery of viable structures in the in vivo production of sheep embryos. In conclusion, there is a median correlation between follicular population observed by ultrasonography and viable recovered structures after superovulation protocol. Therefore, this tool is not indicated as a screening tool, alone, in the selection of Santa Inês sheep embryo donors.

Follicular wave synchronization on in vivo embryo production in Santa Ines sheep.

This study aimed to evaluate two hormonal protocols for synchronization of follicular wave emergence on in vivo embryo production in Santa Ines sheep under tropical conditions. The greater PRCL rate in GT probably contributed to the smaller number of viable embryos. Thus, it is suggested the appliance indicated the GEm protocol for in vivo embryo production in Santa Ines sheep under tropical conditions.

Influence of mini-Percoll techniques in sperm capacitation and plasma membrane integrity of ram frozen-thawed sperm.

2016

Recovery of reproductive activity and fertility of Saanen goats affected by hydrometra after cloprostenol treatment and estrus induction during the non-breeding season (preliminary data).

2016

Reproductive seasonality in Saanen goats managed in southeast Brazil.

The aim of this study was to characterize the reproductive seasonality during the period of one year in Saanen goats managed in southeastern Brazil These results endorse hormonal methods (estrus synchronization and induction of synchronized estrus) or natural (male effect, flushing or light treatment) more appropriate according to the time of year in the region. Saanen goats, managed under tropical climate in southeastern Brazil, have a breeding season set from March until mid-August with the transitional phase to the anestrus in September. It is extends to mid-December at the new one transition phase for the reproductive season in March.

Restricted intake and lipid inclusion: effects on ovarian follicular development in Santa Inês ewe lambs.

The reproductive efficiency is directly related to the nutritional management. The diet lipid inclusion, especially with enhanced sources of omega 6 and omega 3 fatty acids, is positively associated to the ovarian follicular development of ruminants. The objective of this study was to evaluate the effects of toasted soybean grain addition on restricted or ad libitum feeding on ovarian follicular development of Santa Inês ewe lambs In conclusion, the diameter of the largest follicle and the number of >5 mm follicles were not influenced by the restricted intake diet nor the lipid inclusion; however, they increased in a direct relationship with the age of the animals.

Restricted intake and lipid inclusion in Santa Inês ewe lambs diet: age, weight and progesterone concentration at first ovulation.

The age at first ovulation is influenced by several factors, and nutrition has an essential role on it. Lipids provide essential fatty acids that are positively associated to reproductive aspects. The aims of this study were to evaluate the effects of lipid inclusion and restricted intake on age and weight at the first ovulation and the serum progesterone (P4) concentration at the sixth day after first ovulation. The restricted intake imposed in this study did not delay the age at fist ovulation. The greater lipid intake did not favor reproductive parameters. Serum P4 did not increase with the soybean inclusion in the die.