Activation of bovine oocytes by strontium combined or not with an electric pulse

O objetivo deste trabalho foi avaliar as taxas de ativação e de clivagem de oócitos bovinos tratados com estrôncio (10mm de SrCl2), após maturação in vitro por 27-28 horas. No experimento 1, os tratamentos foram: S4 - ativação pelo estrôncio por 4 horas; S12 - ativação pelo estrôncio por 12 horas; S30 - ativação pelo estrôncio por 30 horas; e P - ativação por pulso elétrico (3 pulsos de 1,0kv/cm). No experimento 2 os tratamentos foram: PS4 - ativação combinada pelo pulso elétrico e pelo estrôncio por 4 horas; S4P - ativação pelo estrôncio por 4 horas e pelo pulso elétrico; e PS30 - ativação pelo pulso elétrico e pelo estrôncio por 30 horas. No experimento 1, todos os tratamentos apresentaram taxas similares de ativação (83-90%; P>0,05). Para clivagem, P foi melhor (53%; P<0,05) do que todos os tratamentos com estrôncio (6 a 28%). No experimento 2, o tratamento S4P apresentou melhor taxa de ativação (88%; P<0,05) do que PS4 e PS30 (60 e 68%, respectivamente). Para clivagem, observou-se o mesmo padrão, S4P (65%; P<0,05) e PS4 e PS30 (37% e 44%, respectivamente). Conclui-se que o estrôncio é capaz de ativar oócitos bovinos e sua combinação com pulso elétrico não melhora a ativação. Este é o primeiro relato demonstrando que o estrôncio ativa oócitos bovinos.

Molecular characterization of group A bovine rotavirus in southeastern and central-western Brazil, 2009-2010

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

Avanços metodológicos na seleção do sexo de espermatozóides bovinos para utilização no melhoramento genético e na produção animal

O mercado demanda, desde final da década de 20, uma tecnologia para a sexagem de espermatozóides que possa ser inserida na indústria de produção de sêmen congelado e que tenha as seguintes características: a) não altere a viabilidade espermática; b) seja compatível com a congelação do espermatozóide sexado; c) permita a sexagem de espermatozóides previamente congelados e descongelados; d) permita a produção de várias doses de sêmen sexado congelado por dia, com custo compatível ao mercado. A importância dessa tecnologia para maximizar a produção animal a um custo baixo tem sido um desafio da pesquisa a vários anos. A possibilidade de produzir, em escala comercial, doses de sêmen enriquecidas com espermatozóides X ou Y aumentará os benefícios do uso da inseminação artificial no seu papel de maximizar o progresso genético entre gerações de acordo com os requerimentos de cada programa de melhoramento animal. Diferentes rotas tecnológicas são percorridas na tentativa de selecionar-se o sexo em mamíferos, tanto nas espécies de interesse zootécnico quanto em espécies ameaçadas de extinção, animais de companhia. Neste sentido, existem duas alternativas: a separação de espermatozóides portadores do cromossomo X, daqueles portadores do cromossomo Y; ou a sexagem de embriões pré-implantados. A viabilidade da sexagem de espermatozóides em bovinos é esperada por muitos anos e os desenvolvimentos recentes tornaram essa tecnologia de aplicação commercial. Entretanto, muitas limitações ainda existem, principalmente, referente à taxa de gestação em condições de campo. Isso restringe a utilização dessa tecnologia no melhoramento genético e produção animal. Nessa palestra abordaremos os potenciais sistemas de criação e produção que poderão beneficiar-se com a sexagem de espermatozóides, quando essas limitações forem solucionadas.

Meiotic inhibition of bovine oocytes in medium supplemented with a serum replacer and hormones: effects on meiosis progression and developmental capacity

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

Characterization of bovine transcripts preferentially expressed in testis and with a putative role in spermatogenesis

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

Transmission electron microscopy for characterization of acrosomal damage after Percoll gradient centrifugation of cryopreserved bovine spermatozoa

The objective of this study was to characterize acrosomal ultrastructure following discontinuous Percoll gradient centrifugation of cryopreserved bovine sperm. Semen was collected from six bulls of different breeds and three ejaculates per bull were evaluated. Frozen semen samples were thawed and the acrosomal region of sperm cells was evaluated by transmission electron microscopy (TEM) before (n = 18) and after (n = 18) Percoll centrifugation. The evaluation of 20 sperm heads from each of the 36 samples analyzed ensured that a large number of cells were investigated. The data were subjected to analysis of variance at a level of significance of 5%. Percoll centrifugation reduced the percentage of sperm exhibiting normal acrosomes (from 61.77 to 30.24%), reduced the percentage of sperm presenting atypical acrosome reactions (from 28.38 to 4.84%) and increased the percentage of sperm exhibiting damage in the acrosome (from 6.14 to 64.26%). The percentage of sperm with typical acrosome reactions was not significantly different before (3.70%) and after (0.67%) centrifugation. TEM distinguished four different types of acrosomal status and enabled ultrastructural characterization of acrosomal injuries. The percentage of sperm exhibiting normal acrosomes decreased and damage in the acrosome was the most frequent acrosomal injury with the Percoll gradient centrifugation protocol utilized.

Heat or cold chronic stress affects organ weights and Hsp70 levels in chicken embryos

This study was carried out with the objective of evaluating the effect of heat (38.8 degreesC) or cold (35.8 degreesC) stress on chicken embryo development and tissues Hsp70 levels, after the 13th day of incubation. Embryo weight (percent egg weight), organ weight (percent embryo weight) and Hsp70 levels (ng Hsp70 mug(-1) total protein) in different tissues (liver, breast muscle, heart, lungs, brain and kidney) were studied at the end of incubation. Cold stress induced a lower embryo weight and lower kidney and lungs weights, whereas heart and liver were lighter in heat-stressed embryos. An interaction between temperature and age was obtained only for Hsp70 levels in kidney and heart. Cold-stressed embryos showed higher Hsp70 levels in the brain, lungs and liver; a decrease in brain and breast muscle Hsp70 levels was seen from the 19th to 20th days in control embryos. Hsp70 levels increased with age in kidneys of control embryos and in heart of heat- and cold-stressed embryos. In conclusion, this study showed that chicken embryo organ weights are affected by incubation temperature, and that Hsp70 expression is tissue dependent (higher levels being seen in the brain) being cold-stress more effective in increasing Hsp70 levels in most studied tissues.

Expression of MyoD, myogenin, myostatin and Hsp70 transcripts in chicken embryos submitted to mild cold or heat

The expression of the MyoD, myogenin, myostatin and Hsp70 genes was estimated in chicken embryos submitted to mild cold (36 +/- 0.5degreesC) or heat (44 +/- 0.5degreesC) for 1 h. 2. Marked decreases in MyoD, myogenin and myostatin transcript levels were observed in embryos exposed to high temperature, contrasting to the higher expression of the Hsp70 mRNA detected in heat-stressed embryos. 3. The exposure of chicken embryos to low temperature significantly affected only the abundance of myogenin mRNA. 4. These findings suggest that myogenic proliferation and differentiation events are compromised by variations in environmental temperature during avian embryogenesis. (C) 2003 Elsevier B.V. Ltd. All rights reserved.

Hematological values and body, heart and liver weights of male and female broiler embryos of young and old breeder eggs

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

Breeder age and bone development in broiler chicken embryos

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Effect of breed and corpus luteum on pregnancy rate of bovine embryo recipients

The objective of this study was to evaluate pregnancy rates of recipients of different breed groups (Nellore and crossbreed), as well as the effects of size and type of the corpus luteum (CL) on plasmatic concentrations of progesterone and pregnancy rates of embryo recipients. A total of 152 heifers were synchronized with progesterone implants and on the day of embryo transfer, previously obtained by superovulation and frozen in ethylene glycol, the diameter and type of the corpus luteum (cavitary and compact) was measured and blood was collected for progesterone measurement. The pregnancy rate was 44.1%, with a diameter of corpus luteum higher in recipients that became pregnant (2.03±0.41) compared with non-pregnant ones (1.86±0.34 cm). Plasmatic concentrations of progesterone did not differ between pregnant (1.50±1.05) and non-pregnant (1.31±0.91 ng/mL) animals. The type of corpus luteum did not influence the pregnancy rates. Only Angus and crossbred Marchigiana differ among themselves in pregnancy rates (33.3 and 59.2%, respectively). The pregnancy probability was affected only by CL diameter, but not by P4 plasmatic concentration. Selection of the corpus luteum size at the time of embryo transfer is an important factor to increase pregnancy rates in recipients, and compact and cavitary corpora lutea do not influence the pregnancy rates of bovine embryo recipients. Nellore recipients have pregnancy rates that are satisfactory and comparable to crossbred (Bos taurus × Bos indicus) recipients.

Possible role of bovine trophoblast giant cells in transplacental transmission of Neospora caninum in cattle

Neospora caninum is an aplicomplexan parasite that has brought several concerns to cattle raisers worldwide due to its relationship to fetal loss. However, the mechanism of the parasite's transplacental infection and induced abortions are not completely understood. Bovine trophoblastic binucleated cells (BNC) play a major role in the maternal-fetal interactions, migrating during the entire pregnancy from chorionic connections to uterine epithelium. This study aimed to investigate the possible role of BNC as phagocytic cells and its participation in the bovine transplacental infection of N. caninum. BNC was isolated by discontinuous Percoll gradient, and characterized by Hoeschst 33342 nucleus-specific staining. Isolated BNC were cultured in DMEM supplemented with 10% bovine fetal serum, and infected with 10(4) tachyzoites of N. caninum NC-1 strain. Parasite invasion was visualized by indirect immunofluorescence and Giemsa technique. Multiplication of parasites took place in 2-3 day cycles. Healthy cows' placenta and normal and infected cultured BNC was immunostained with monoclonal antibodies against CD-163, MAC-387 and NOS, demonstrating their phagocyte capacity. Thus, BNC was characterized as cells with macrophagic activity, which may host N. caninum in vitro. Therefore, we may conclude that BNC could potentially participate in the transplacental infection of bovine neosporosis.

Systemic and local antibodies induced by an experimental inactivated vaccine against bovine herpesvirus type 1

Uma vacina experimental inativada contra o herpesvírus bovino tipo 1 (BoHV-1) foi produzida com o objetivo de se avaliar a resposta imune humoral local e sistêmica contra o BoHV-1, em 12 novilhas soronegativas, após a vacinação e a revacinação. Os soros foram submetidos à prova de vírus-neutralização para quantificação do título de anticorpos neutralizantes e a um ELISA para detecção de IgG1 e IgG2. Os swabs nasais também foram submetidos ao ELISA para detecção de IgG1 e IgG2 na secreção nasal. Os resultados demonstraram que títulos de anticorpos neutralizantes foram induzidos após a revacinação, em níveis moderados a altos, permanecendo em níveis significativos no soro sanguíneo e na secreção nasal até o dia 114 pós-vacinação. O IgG2 foi o isótipo predominante na maior parte do período pós-vacinação, tanto na secreção nasal, como no compartimento sistêmico. A vacina experimental inativada contra o BoHV-1 estimulou níveis de anticorpos potencialmente protetores dos isótipos IgG1 e IgG2, tanto no compartimento sistêmico, como nas mucosas.

Detection of bovine herpesvirus 5 (BoHV-5) in formalin-fixed, paraffin-embedded bovine brain by nested PCR in Colombian cattle

Fifteen cases of viral meningoencephalitis in Colombian cattle were tested by nested PCR analysis for the detection of bovine herpesvirus 5 (BoHV-5). All fatal cases had shown severe neurological signs and had occurred following natural outbreaks of the disease. The neurological infection was histologically characterized by mild to moderate inflammatory changes in the brain and cerebellum, including meningitis, mononuclear perivascular cuffing, gliosis, haemorrhage, and the presence of Gitter cells (macrophages) accompanying large areas of malacia. No intranuclear inclusion bodies were seen in any of the cases. Results from BoHV-5 molecular extraction analyses showed there were five positive cases thus confirming the presence of the virus in Colombia.

Rhipicephalus (Boophilus) microplus: Distinct acute phase proteins vary during infestations according to the genetic composition of the bovine hosts, Bos taurus and Bos indicus

Tick bites may trigger acute phase responses. Positive and negative acute phase proteins were measured in infested cattle genetically resistant and susceptible to ticks. During heavier infestations levels of haptoglobin increased significantly in susceptible bovines; levels of serum amyloid A increased in resistant bovines; levels of alpha-l-acid glycoprotein decreased significantly in resistant bovines; levels of transferrin decreased significantly in susceptible bovines.In conclusion, tick infestations trigger acute phase responses and enhancement of specific acute phase proteins differs according to the genetic composition of hosts. Acute phase proteins may constitute useful biological signatures for monitoring the stress induced by tick infestations. (c) 2007 Elsevier B.V. All rights reserved.