Dynamics of DNA Methylation during Early Development of the Preimplantation Bovine Embryo

There is species divergence in control of DNA methylation during preimplantation development. The exact pattern of methylation in the bovine embryo has not been established nor has its regulation by gender or maternal signals that regulate development such as colony stimulating factor 2 (CSF2). Using immunofluorescent labeling with anti-5-methylcytosine and embryos produced with X-chromosome sorted sperm, it was demonstrated that methylation decreased from the 2-cell stage to the 6-8 cell stage and then increased thereafter up to the blastocyst stage. In a second experiment, embryos of specific genders were produced by fertilization with X- or Y-sorted sperm. The developmental pattern was similar to the first experiment, but there was stage × gender interaction. Methylation was greater for females at the 8-cell stage but greater for males at the blastocyst stage. Treatment with CSF2 had no effect on labeling for DNA methylation in blastocysts. Methylation was lower for inner cell mass cells (i.e., cells that did not label with anti-CDX2) than for trophectoderm (CDX2-positive). The possible role for DNMT3B in developmental changes in methylation was evaluated by determining gene expression and degree of methylation. Steady-state mRNA for DNMT3B decreased from the 2-cell stage to a nadir for D 5 embryos >16 cells and then increased at the blastocyst stage. High resolution melting analysis was used to assess methylation of a CpG rich region in an intronic region of DNMT3B. Methylation percent decreased between the 6-8 cell and the blastocyst stage but there was no difference in methylation between ICM and TE. Results indicate that DNA methylation undergoes dynamic changes during the preimplantation period in a manner that is dependent upon gender and cell lineage. Developmental changes in expression of DNMT3B are indicative of a possible role in changes in methylation. Moreover, DNMT3B itself appears to be under epigenetic control by methylation. © 2013 Dobbs et al.

Optimal single-embryo mass spectrometry fingerprinting

In pre-implantation embryos, lipids play key roles in determining viability, cryopreservation and implantation properties, but often their analysis is analytically challenging because of the few picograms of analytes present in each of them. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) allows obtaining individual phospholipid profiles of these microscopic organisms. This technique is sensitive enough to enable analysis of individual intact embryos and monitoring the changes in membrane lipid composition in the early stages of development serving as screening method for studies of biology and biotechnologies of reproduction. This article introduces an improved, more comprehensive MALDI-MS lipid fingerprinting approach that considerably increases the lipid information obtained from a single embryo. Using bovine embryos as a biological model, we have also tested optimal sample storage and handling conditions before the MALDI-MS analysis. Improved information at the molecular level is provided by the use of a binary matrix that enables phosphatidylcholines, sphingomyelins, phosphatidylserines, phosphatidylinositols and phosphoethanolamines to be detected via MALDI(±)-MS in both the positive and negative ion modes. An optimal MALDI-MS protocol for lipidomic monitoring of a single intact embryo is therefore reported with potential applications in human and animal reproduction, cell development and stem cell research. Copyright © 2013 John Wiley & Sons, Ltd. Copyright © 2013 John Wiley & Sons, Ltd.

Effect of dietary betaine supplementation on the performance, carcass yield, and intestinal morphometrics of broilers submitted to heat stress

The objective of this study was to evaluate the effect of betaine in methionine- and choline-reduced diets fed to broilers submitted to heat stress. In total, 1,408 male broilers were randomly distributed into eight treatments, according to 2 × 4 (environment x diet) factorial arrangement, with eight replicates of 2 birds each. Birds were reared environmental chambers under controlled temperature (25-26 °C) or cyclic heat-stressing temperature (25-31 °C). The following diets were tested: positive control (PC), formulated to meet broiler nutritional requirements; negative control (NC), with reduced DL-methionine and choline chloride levels; and with two supplementation levels of natural betaine to the negative control diet (NC+NB1 and NC+NB2). Live performance, carcass traits, and intestinal morphometrics were evaluated when broilers were 45 days of age. The results showed that all evaluated parameters were influenced by the interaction between environment and diet, except for breast meat drip loss. The breakdown of the interactions showed that birds fed the PC diet and reared in the controlled environment had greater breast drip loss than those submitted to the cyclic heat-stress environment. Birds submitted to cyclic heat stress and fed the PC diet presented the lowest feed intake. Feed conversion ratio was influenced only by diet. The FCR of broilers fed the NC+NB2 diet was intermediate relative to those fed the PC and NC diets. The addition of betaine in the diet, with 11.18% digestible methionine and 24.73% total choline reductions, did not affect broiler live performance, carcass yield, or intestinal morphometrics.

Genetic parameters for milk yield, lactation length and calving intervals of murrah buffaloes from brazil

The major objective of this study was to estimate heritability and genetic correlations between milk yield (MY) and calving interval (CI) and lactation length (LL) in Murrah buffaloes using Bayesian inference. The database used belongs to the genetic improvement program of four buffalo herds from Brazil. To obtain the estimates of variance and covariance, bivariate analyses were performed with the Gibbs sampler, using the program MTGSAM. The heritability coefficient estimates were 0.28, 0.03 and 0.15 for MY, CI and LL, respectively. The genetic correlations between MY and LL was moderate (0.48). However, the genetic correlation between MY and CI showed large HPD regions (highest posterior density interval). Milk yield was the only trait with clear potential for genetic improvement by direct mass selection. The genetic correlation between MY and LL indicates that indirect selection using milk yield is a potentially beneficialstrategy.Theinterpretation of the estimated genetic correlation between MY and CI is difficult and could be spurious. ©2013 Sociedade Brasileira de Zootecnia.

Multiple-trait random regression models for the estimation of genetic parameters for milk, fat, and protein yield in buffaloes

In this study, genetic parameters for test-day milk, fat, and protein yield were estimated for the first lactation. The data analyzed consisted of 1,433 first lactations of Murrah buffaloes, daughters of 113 sires from 12 herds in the state of São Paulo, Brazil, with calvings from 1985 to 2007. Ten-month classes of lactation days were considered for the test-day yields. The (co)variance components for the 3 traits were estimated using the regression analyses by Bayesian inference applying an animal model by Gibbs sampling. The contemporary groups were defined as herd-year-month of the test day. In the model, the random effects were additive genetic, permanent environment, and residual. The fixed effects were contemporary group and number of milkings (1 or 2), the linear and quadratic effects of the covariable age of the buffalo at calving, as well as the mean lactation curve of the population, which was modeled by orthogonal Legendre polynomials of fourth order. The random effects for the traits studied were modeled by Legendre polynomials of third and fourth order for additive genetic and permanent environment, respectively, the residual variances were modeled considering 4 residual classes. The heritability estimates for the traits were moderate (from 0.21-0.38), with higher estimates in the intermediate lactation phase. The genetic correlation estimates within and among the traits varied from 0.05 to 0.99. The results indicate that the selection for any trait test day will result in an indirect genetic gain for milk, fat, and protein yield in all periods of the lactation curve. The accuracy associated with estimated breeding values obtained using multi-trait random regression was slightly higher (around 8%) compared with single-trait random regression. This difference may be because to the greater amount of information available per animal. © 2013 American Dairy Science Association.

Comparison of factor-analytic and reduced rank models for test-day milk yield in Gyr dairy cattle (Bos indicus)

We analyzed 46,161 monthly test-day records of milk production from 7453 first lactations of crossbred dairy Gyr (Bos indicus) x Holstein cows. The following seven models were compared: standard multivariate model (M10), three reduced rank models fitting the first 2, 3, or 4 genetic principal components, and three models considering a 2-, 3-, or 4-factor structure for the genetic covariance matrix. Full rank residual covariance matrices were considered for all models. The model fitting the first two principal components (PC2) was the best according to the model selection criteria. Similar phenotypic, genetic, and residual variances were obtained with models M10 and PC2. The heritability estimates ranged from 0.14 to 0.21 and from 0.13 to 0.21 for models M10 and PC2, respectively. The genetic correlations obtained with model PC2 were slightly higher than those estimated with model M10. PC2 markedly reduced the number of parameters estimated and the time spent to reach convergence. We concluded that two principal components are sufficient to model the structure of genetic covariances between test-day milk yields. © FUNPEC-RP.

Yield and mineral nutrition of soybean, maize, and congo signal grass as affected by limestone and slag

The objective of this work was to evaluate the efficiency of superficial applicationof limestone and slag, and their effects on soil chemical attributes and on yield and mineral nutrition of soybean, maize, and Congo signal grass (Urochloa ruziziensis). The experiment was carried out in a Rhodic Hapludox under no tillage system. The treatments consisted of the use of limestone or slag (silicates of calcium and magnesium) to correct soilacidity, and of a control treatment without the use of soil correctives. Rates were calculated in order to raise soil base saturation up to 70%. Soybean was sown in November 2006and maize in December 2007. Congo signal grass was sown right after the harvests of soybean and maize, and it was cropped during the offseasons. Soil chemical attributes were evaluated at 6, 12, and 18 months after the application of the corrective materials. Slag isan efficient source for soil acidity correction, being able to raise the exchangeable base levels in the soil profile faster than lime. Both limestone and slag increase dry matter yield of Congo signal grass, and grain yield of soybean and maize. Slag is more effective in improving maize grain yield.

Sorghum grain yield, forage biomass production and revenue as affected by intercropping time

Sorghum is an excellent alternative to other grains in poor soil where corn does not develop very well, as well as in regions with warm and dry winters. Intercropping sorghum [Sorghum bicolor (L.) Moench] with forage crops, such as palisade grass [Brachiaria brizantha (Hochst. ex A. Rich) Stapf] or guinea grass (Panicum maximum Jacq.), provides large amounts of biomass for use as straw in no-tillage systems or as pasture. However, it is important to determine the appropriate time at which these forage crops have to be sown into sorghum systems to avoid reductions in both sorghum and forage production and to maximize the revenue of the cropping system. This study, conducted for three growing seasons at Botucatu in the State of São Paulo in Brazil, evaluated how nutrient concentration, yield components, sorghum grain yield, revenue, and forage crop dry matter production were affected by the timing of forage intercropping. The experimental design was a randomized complete block design. Intercropping systems were not found to cause reductions in the nutrient concentration in sorghum plants. The number of panicles per unit area of sorghum alone (133,600), intercropped sorghum and palisade grass (133,300) and intercropped sorghum and guinea grass (134,300) corresponded to sorghum grain yields of 5439, 5436 and 5566kgha-1, respectively. However, the number of panicles per unit area of intercropped sorghum and palisade grass (144,700) and intercropped sorghum and guinea grass (145,000) with topdressing of fertilizers for the sorghum resulted in the highest sorghum grain yields (6238 and 6127kgha-1 for intercropping with palisade grass and guinea grass, respectively). Forage production (8112, 10,972 and 13,193Mg ha-1 for the first, second and third cuts, respectively) was highest when sorghum and guinea grass were intercropped. The timing of intercropping is an important factor in sorghum grain yield and forage production. Palisade grass or guinea grass must be intercropped with sorghum with topdressing fertilization to achieve the highest sorghum grain yield, but this significantly reduces the forage production. Intercropping sorghum with guinea grass sown simultaneously yielded the highest revenue per ha (€ 1074.4), which was 2.4 times greater than the revenue achieved by sowing sorghum only. © 2013 Elsevier B.V.

Nitrogen and potassium fertilization on the yield and intensity of the maize white spot

A plant’s nutritional balance can influence its resistance to diseases. In order to evaluate the effect of increasing doses of N and K on the yield and severity of the maize white spot, two experiments were installed in the field, one in the city of Ijaci, Minas Gerais, and the other in the city of Sete Lagoas, Minas Gerais. The experimental delimitation was in randomized blocks with 5 x 5 factorial analysis of variance, and four repetitions. The treatments consisted of five doses of N (20; 40; 80; 150; 190 Kg ha-1 of N in the experiments 1 and 2) and five doses of K (15; 30; 60; 120; 180 Kg ha-1 of K in experiment 1 and 8.75; 17.5; 35; 50; 100 Kg ha-1 of K in experiment 2). The susceptible cultivar 30P70 was planted in both experiments. The plot consisted of four rows 5 meters long, with a useful area consisting of two central rows 3 meters each. Evaluations began 43 days after emergence (DAE) in the first experiment and 56 DAE in the second one. There was no significant interaction between doses of N and K and the disease progress. The effect was only observed for N. The K did not influence the yield and the severity of the disease in these experiments. Bigger areas below the severity progress curve of the white spot and better yield were observed with increasing doses of N. Thus, with increasing doses of N, the white spot increased and also did the yield.

Fatores que afetam a taxa de concepção após inseminação artificial ou transferência de embriões em vacas holandesas em lactação

Pós-graduação em Medicina Veterinária - FMVZ

Simulation of soybean growth and yield under northeastern Amazon climatic conditions

O objetivo deste trabalho foi parametrizar, calibrar e validar uma nova versão do modelo de crescimento e de produtividade da soja desenvolvido por Sinclair, em condições naturais de campo no nordeste da Amazônia. Os dados meteorológicos e os valores de crescimento e de área foliar da soja foram obtidos em um experimento agrometeorológico realizado em Paragominas, PA, de 2006 a 2009. As condições climáticas durante o experimento foram muito distintas, com uma ligeira redução na precipitação em 2007, em virtude do fenômeno El Niño. Houve redução no índice de área foliar (IAF) e na produção de biomassa neste ano, a qual foi reproduzida pelo modelo. A simulação do IAF apresentou raiz do erro quadrado médio (REQM) de 0,55 a 0,82 m2 m‑2, de 2006 a 2009. A simulação da produtividade da soja para os dados independentes apresentou um REQM de 198 kg ha‑1, ou seja, uma superestimativa de 3%. O modelo encontra-se calibrado e validado para as condições climáticas da Amazônia e pode contribuir positivamente para a melhoria das simulações dos impactos da mudança de uso da terra na região amazônica. A versão modificada do modelo de Sinclair simula adequadamente a formação de área foliar, a biomassa total e a produtividade da soja, nas condições climáticas do nordeste da Amazônia.

Morfoanatomia de ovário, pericarpo e semente de sete espécies de Myrteae DC. (Myrtaceae)

Pós-graduação em Ciências Biológicas (Botânica) - IBB

Biomass yield and calorific value of six clonal stands of Eucalyptus urophylla S. T. Blake cultivated in Northeastern Brazil

A demanda mundial por energia limpa e sustentável tem aumentado nos últimos anos. Uma das alternativas é a biomassa, incluindo os plantios florestais, com destaque, no Brasil, para o gênero Eucalyptus spp., que, atualmente, possui diversos clones selecionados para a produção de energia. Em razão da diferença na adaptação dos clones aos diferentes ambientes, há necessidade de estudos específicos para avaliar a produção de energia para determinada região. Objetivou-se, neste estudo, determinar dentre seis clones de Eucalyptus urophylla S. T. Blake analisados, aqueles com maior potencial para a produção de biomassa e energia para a região Centro Sul do Maranhão (Grajaú). Os clones foram avaliados com 41 meses de idade, plantados em espaçamento de 4 x 3 m, e distribuídos em delineamento inteiramente casualizado, com cinco repetições. Cada parcela experimental era composta por 100 árvores. Foram determinadas a produção e a alocação de biomassa de cada parte da árvore e o poder calorífico de casca e do lenho. Dos seis clones estudados, dois se destacaram tanto na produção de biomassa, quanto para a geração de energia, sendo superiores em até 27 % em comparação com o clone de menor rendimento.

Genetic parameters and simultaneous selection for root yield, adaptability and stability of cassava genotypes

O objetivo deste trabalho foi estimar parâmetros genéticos e avaliar a seleção simultânea quanto à produtividade de raízes e à adaptabilidade e estabilidade de genótipos de mandioca. Os efeitos dos genótipos foram considerados como fixos e aleatórios, e a metodologia de modelos mistos (REML/Blup) foi utilizada para estimar os parâmetros genéticos e a média harmônica do desempenho relativo dos valores genotípicos (MHPRVG), para seleção simultânea. Dez genótipos foram avaliados em delineamento de blocos ao acaso, com quatro repetições. O experimento foi realizado nos municípios de Altamira, Santarém e Santa Luzia do Pará, PA, nos anos agrícolas de 2009/2010, 2010/2011 e 2011/2012. As raízes foram colhidas 12 meses após o plantio, em todos os locais testados. A produtividade de raízes apresentou baixo coeficiente de variação genotípica (4,25%) e herdabilidade de parcelas individuais no sentido amplo (0,0424), o que resultou em baixo ganho genético. Em razão da baixa correlação genotípica (0,15), a classificação dos genótipos quanto à produtividade de raízes variou de acordo com o ambiente. Os genótipos CPATU 060, CPATU 229 e CPATU 404 destacaram-se quanto à produtividade, adaptabilidade e estabilidade.

Evaluation of Meteorological Elements and Lettuce (lactuca Sativa, L.) Yield Related to Geographic Orientation of Polyethylene Greenhouses

O experimento foi realizado na área experimental do Departamento de Engenharia Rural da Faculdade de Ciências Agronômicas – Unesp/Botucatu, Estado de São Paulo, em duas estufas dispostas em diferentes orientações geográficas, Leste/Oeste e Norte/Sul. A alface (Lactuva sativa L.) cv. Elisa foi cultivada em ambas estufas, sendo semeada em 05/05/99, transplantada em 29/05/99 e colhida em 31/06/99. Utilizou-se tensiômetros para monitorar o potencial de água no solo para realizar o manejo do sistema de irrigação por gotejamento. Microevaporímetros eqüidistantes de 3 m e colocados em 3 alturas, 0.50, 1.00 e 1.50 m, termohigrógrafos e tanques Classe “A” foram instalados nas duas estufas. Através de análise geoestatística, não se observou dependência espacial nem variabilidade espacial da evaporação nas duas estufas. Entretanto, a altura dos evaporímetros apresentou diferenças significativas: a evaporação à altura de 1.50 foi menor que nas outras duas.As médias de temperatura, umidade relativa e déficit de pressão de vapor do arnão diferiram estatisticamente entre as estufas e o ambiente externo. Os valores médios de evaporação de água no tanque Classe A instalado fora das estufas diferiram estatisticamente quando comparados com os instalados no interior das estufas, porém, entre as orientações não se constatou diferença significativa. Pôde-se verificar que não houve diferença significativa das características agronômicas da alface em ambas orientações estudadas. No entanto, houve diferença significativa para essas características entre os canteiros no interior das mesmas, havendo variância espacial para os dados de matéria fresca apenas na estufa N/S.