Técnicas associadas à biotecnologia da reprodução para minimizar os efeitos do estresse térmico e aumentar a fertilidade em vacas leiteiras de alta produção

The dairy sector has undergone considerable economic losses due to low fertility rates due to adverse effects of heat stress on reproduction of cows. Genetic selection for increased production, coupled with the expanding dairy to tropical areas of the planet, and global warming has further aggravated the problem of heat stress. The effects of heat stress are multifactorial and act directly or indirectly at various levels of reproductive tissues, resulting in low fertility of cows, which in practice, results in reduced reproductive efficiency in the property, reducing the producers’ profit. Some strategies related to breeding biotechnology such as fixed-time artificial insemination, embryo transfer and use of BST, can minimize these effects and improve the reproductive efficiency of the herd

Principais marcadores utilizados na avaliação da qualidade do sêmen de animais domésticos

Methods of semen cryopreservation allow changes in spermatic cells, such as damage in plasma and acrossomal membrane and modifications in mitochondrial function due to a disorder in the lipidic bilayer. For effective oocyte fertilization, spermatozoa require functional competent membranes, and intact organelles, acrosome and DNA. However, most laboratory methods used to evaluate semen quality are not highly correlated with fertilizing capacity. The discovery of a variety of fluorochromes and compounds conjugated to fluorescent probes has enabled an accurate assessment of the viability, integrity and function of spermatozoa. Among the most used probes that label the various compartments of the sperm cell there are the membrane impermeable fluorescent dyes to test the membrane integrity, as well as acylated dyes that pass the intact membrane. For the acrossomal integrity the most commonly used method is lectins labeled by a fluorescent probe. The acrosome reaction and spermatic capacitation is detected by the evaluation of membrane architecture and disorder of lipids in plasma membrane. Mitochondrial function can be determined using markers for their aerobic activity. The DNA status of spermatozoa has been determined using the metachromatic properties of Acridine Orange, and the DNA fragmentation can also be assessed by TUNEL assay. Finally, DNA condensation is analyzed using a single cell DNA gel electrophoresis assay that indicates DNA compactation. This monograph aims to compile the various tests used to detect damaged spermatozoa under cryopreservation methods, searching for improve the predictive value of semen analysis with the intention of a successful conception

Dinâmica folicular ovariana e bases para a aplicação das biotécnicas da reprodução em bovinos

In the last years Brazil has achieved a leading position in worldwide cattle breeding. This country is consolidating the practice of an intensive and technical level cattle breeding, due to greater emphasis on reproductive efficiency and genetic improvement. The attention given to reproductive efficiency became to enhance the genetic improvement process, by exploiting the full reproductive potential of the animals. The biotechniques applied to reproduction arised to expand the reproductive potential for greater economic return. Are the most important reproductive biotechniques the FTAI and in vivo and in vitro embryo production for embryo transfer. Different hormonal treatments are consolidating to interfere with normal physiology in order to achieve the desired goals. The FTAI works to increase the reproductive rates of cattle through pre-booked artificial insemination in all the females in reproduction, without the need for estrous detection. The in vivo and in vitro embryo production aims to distribute the genetics of males and females genetically improved by transferring their embryos to genetically inferior females. Superovulation treatments designed to prevent the mechanism of dominance, which results in many follicles ovulating simultaneously for in vivo embryo production. To the in vitro embryo production, the technique of ovarian puncture guided by transvaginal ultrasound allows to aspirate a number of quality oocytes. The application of biotech is based on knowledge of the female and ovarian reproductive physiology and knowledge of their acting in different breeds and environments.

Fatores que podem influenciar o sexo da cria em bovinos

To assess the possibility of shifting the sex ratio at birth, since this can contribute to enhancing the genetic gain and producity in cattle, this work is aimed at raising the factors that may influence sex determination of creates. This study is based on research conducted in the area of reproduction and production of beef and dairy cattle, some quotes in humans, mouse, dogs and a study in pigs. This was due to back of data and studies in the bovine species. It was noted that stress during pregnancy, intake of vitamin C or others substances and the time of artificial insemination are the factors that may influence the determination of the type of product. Human studies, concluded that woman more stressed are more likely to produce female children of the quietest. Furthermore, sows that received ascorbic a ad orally for seven days and were inseminated during this period, produce more female then those who did not receive the vitamin. These studies may suggest that cows can also suffer influence of stress and food for the determination of sex of calf. There are also studies suggesting that cows inseminated at the earliest time of ovulation ted to produce more male calves than those inseminated a few hours before that time. Some scholars considers the hypothesis... (Complete abstract click electronic access below)

Indução de parto em bovinos: Princípios e aplicações

Currently, Brazil has one of the largest cattle herds worldwide. In order to keep that milk and meat were introduced reproductive biotechnologies such as artificial insemination, embryo transfer and in vitro fertilization. In certain situations the technique may have undesired effect, for example, the production of calve calves due to the very large increase in the gestation period when performed in vitro fertilization. To avoid this problem we perform the induction of labor in order to prevent the product is longer the womb. This induction can also be made in case of diseases that compromise the life of the mother, twin pregnancy an abnormal size calf. The administration of short acting steroids, prostaglandins, association of short acting steroids and prostaglandins and association of short acting steroids, prostaglandins and long-acting corticosteroids are some of the possibilities of induction

Fatores que interferem nas taxas de recuperação embrionária em éguas

The success in implementing an embryo transfer program derives from many factors. The embryo recovery rate is one of the most important factors, and allows the transfer to the recipient mare. This rate comes from a group of elements such as donor´s characteristics, collection date, reproductive management, semen quality, technician skill. Mainly because of the great importance of this direct interference on the results of an embryo recovery program, this study aimed to review the factors involved in embryo recovery rates in donor mares during an embryo transfer, suggesting some ways of improving these results

Resposta endometrial de jumentas (Equus asinus) da raça Pêga ao sêmen fresco, extendido e congelado de jumento

Pós-graduação em Cirurgia Veterinária - FCAV

Toxicologia reprodutiva de ratos adultos expostos ao 2,3,7,8-Tetraclorodibenzo-P-Dioxina (TCDD) in útero

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Como reduzir o intervalo entre partos de vacas nelore (bos taurus indicus) e qual sua importância para a pecuária brasileira?

A bovinocultura é um dos principais destaques dentro do universo pecuário brasileiro e mundial, uma vez que o rebanho nacional é detentor do expressivo número de 200 milhões de cabeças entre as atividades de corte e leite, produzindo um valor bruto estimado em 67 bilhões de reais. Ainda assim, a pecuária de corte nacional apresenta baixos índices produtivos por continuar sendo conduzida, ao menos em considerável parcela do país, como uma atividade extrativista que emprega reduzido uso de insumos e biotecnologias, como inseminação artificial, transferência de embriões, fertilização in vitro e inseminação artificial em tempo fixo. No atual cenário mundial, essa característica da bovinocultura brasileira necessita ser alterada para que a atividade torne-se ainda mais rentável e competitiva. Neste caso, a pecuária de corte nacional terá que se apoiar no aumento de sua eficiência reprodutiva e produtiva, tendo como principais alternativas para atingir tal objetivo a elevação do número de vacas com estro no início da estação de monta, o aumento na taxa de concepção no primeiro serviço e a redução no intervalo entre partos, de improdutivos 21 meses para um ano, tempo considerado ideal e de máxima eficiência reprodutiva de um rebanho. Assim sendo, torna-se imprescindível elencar os principais fatores prolongadores do intervalo entre partos para, somente depois, instituir um ou mais métodos para se reduzir essa eficiência reprodutiva e finalmente impulsionar ainda mais a pecuária bovina brasileira

Proliferação celular em gestações naturais e de conceptos bovinos transgênicos e clonados, que expressam a proteína fluorescente verde (GFP)

Pós-graduação em Ciência e Tecnologia Animal - FEIS

Uso de LH ou eCG no último dia do protocolo superovulatório em bovinos

In the last decades several hormonal treatments to induce multiple ovulation and embryo transfer (MOET) have been developed. Tight control of the time of ovulation allowed the use of fixed-time artificial insemination (FTAI) in bovine embryos donors, facilitating animal management. Although, protocols that allow FTAI have evolved and yield as much embryo as conventional protocols that requires estrus detection, substantial increase in viable embryo production has not been observed in superestimulated bovine cattle. The present review put emphasis on superestimulatory protocols in wich the last two doses of pFSH are replaced by eCG or LH. Recent results indicate that an extra LH stimulus (using eCG or LH), on the last day of P-36 superestimulatory treatment, seems to improve transferable embryo yield in both Bos taurus and Bos indicus cattle.

Centrifugação de espermatozóides ovinos em gradiente de densidade contínuo: efeito sobre a taxa de prenhez e proporção do sexo

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Impaired protamination and sperm DNA damage in a Nellore bull with high percentages of morphological sperm defects in comparison to normospermic bulls

The routine semen evaluation assessing sperm concentration, motility and morphology, does not identify subtle defects in sperm chromatin architecture. Bulls appear to have stable chromatin, with low levels of DNA fragmentation. However, the nature of fragmentation and its impact on fertility remain unclear and there are no detailed reports characterizing the DNA organization and damage in this species. The intensive genetic selection, the use of artificial insemination and in vitro embryo production associated to the cryopreservation process can contribute to the chromatin damage and highlights the importance of sperm DNA integrity for the success of these technologies. Frozen-thawed semen samples from three ejaculates from a Nellore bull showed high levels of morphological sperm abnormalities (55.8±5.1%), and were selected for complementary tests. Damage of acrosomal (76.9±8.9%) and plasma membranes (75.7±9.3%) as well as sperm DNA strand breaks (13.8±9.5%) and protamination deficiency (3.7±0.6%) were significantly higher compared to the values measured in the semen of five Nellore bulls with normospermia (24.3±3.3%; 24.5±6.1%; 0.6±0.5%; 0.4±0.6% for acrosome, plasma membrane, DNA breaks and protamine deficiency, respectively) (P<0.05). Motility and percentage of spermatozoa with low mitochondrial potential showed no differences between groups. This study shows how routine semen analyses (in this case morphology) may point to the length and complexity of sperm cell damage emphasizing the importance of sperm function testing.

Efeitos da somatotropina recombinante bovina na taxa de concepção, composição celular do corpo lúteo e morfometria das glândulas endometriais em vacas de corte

Pós-graduação em Ciência Animal - FMVA

Caracterização das proteínas caveolinas -1 e -2 na placenta de conceptos bovinos clonados transgênicos

The transgenic application of green fluorescent protein (GFP) as fetal cell marker on cattle cloned placenta could provide an exclusive model for studying the morphologic and immunologic maternal-fetal interactions, providing information about its mapping, distinguishing the fetal from maternal cells. This model will have direct application, mainly because these animals present problems during its development. With this model's support, we intend to verify the substances transport between mother and fetus during endocytosis, through the immunolocalization of protein named caveolae. For these, we used 06 cloned bovine and 30 cattle samples of artificial insemination (AI) with 90 days of pregnancy, which had been their development interrupted by humanitarian slaughter of the recipient and recovery of the pregnant uterus. We collected the placentome and the chorion. A part of the samples was cut and fixed, by immersion, on a solution containing 4% of parafomaldehyde or 10% of formaldehyde on a sodium phosphate buffer (PBS), at 0,1 M pH 7.4, Zamboni solution (4% of paraformaldehyde, 15% of picric acid, on sodium phosphate buffer 0,1 M pH 7.4), metacarn (60% of metanol, 30% of chloroform, and 10% glacial acetic acid), for morphologic and immunohistochemistry verification for caveolinas proteins -1 and -2 (CAV -1 and CAV-2). The caveolins -1 were found in fetal and maternal villi, but its strongest staining was observed in the endometrial stroma. The caveolins -2 had positive staining in trophoblast and chorioallantoic membrane, and specifically in giant trophoblastic binucleated cell. Therefore the results were compared between cloned cattle and from AI or natural mating, for assisting on detection of the reason of many placental alterations, embryonic losses, spontaneous abortion, post-natal mortality and large offspring syndrome on laboratory-manipulated animals. The result suggests that the proteins caveolins -1 and -2 (CAV-1 and CAV-2) are part of the caveolae composition and important structures related to the molecule transfer to the fetus, nourish it through endocytosis and pinocytosis.