Energy and phosphorus values of sunflower meal and rice bran for broiler chickens using the regression method.

The energy and phosphorus values of sunflower meal (SFM) and rice bran (RB) were determined in 2 experiments with Ross 708 broiler chickens from 15 to 22 d of age. In Exp.1, the diets consisted of a corn-soybean meal reference diet (RD) and 4 test diets (TD). The TD consisted of SFM and RB that partly replaced the energy sources in the RD at 100 or 200 g/kg and 75 or 150 g/kg, respectively, such that the equal ratios were maintained for all energy containing ingredients across all experimental diets. In Exp.2, a cornstarch-soybean meal diet was the RD and TD consisting of SFM and RB that partly replaced cornstarch in the RD at 100 or 200 g/kg and 60 or 120 g/kg, respectively. Addition of SFM and RB to the RD in Exp.1 linearly decreased (P < 0.01) the digestibility coefficients of DM, energy, ileal digestible energy (IDE), metabolizability coefficients of DM, nitrogen (N), energy, N correct energy, metabolize energy (ME), and nitrogen-corrected ME. Except for RB, the increased levels of the test ingredients in RD did affect the metabolizability coefficients of N. The IDE values (kcal/kg DM) were 1,953 for SFM and 2,498 for RB; ME values (kcal/kg DM) were 1,893 for SFM and 2,683 for RB; and MEn values (kcal/kg DM) were 1,614 for SFM and 2,476 for RB. In Exp.2, there was a linear relationship between phosphorus (P) intake and ileal P output for diets with increased levels of SFM and RB. In addition, there was a linear relationship between P intake and P digestibility and retention for diets with increased levels of SFM. There were a quadratic effect (P < 0.01) and a tendency of quadratic effect (P = 0.07) for P digestible and total tract P retained, respectively, in the RB diets. The P digestibility and total tract P retention from regression analyses for SFM were 46% and 38%, respectively.

Identification of the transcriptionally active cytochrome P450 repertoire in Coffea arabica.

Cytochrome P450s (P450s) comprise a gene superfamily encoding enzymes that are involved in diverse plant metabolic pathways that produce primary and secondary metabolites such as phenylpropanoids, terpenoids, nitrogen-containing compounds, and plant hormones. They comprise one of the most diverse gene families in plant evolution. Although there are many studies that aim to characterize P450s in plants, there is no report on the characterization of this superfamily in Coffea arabica, where they might be related to plant tolerance to biotic and abiotic stresses, as well as aroma-related compounds. In this study, we report the characterization and annotation of 87 putative P450s from C. arabica obtained from the Brazilian Coffee Genome Project and describe their transcriptional pattern in different tissues and coffee organs. To validate our approach, we measured the transcriptional profile of the CaCYP81D8_1 gene by quantitative polymerase chain reaction in leaves, flowers, and fruits. This study is the first effort to present and analyze the P450 superfamily in C. arabica, which may assist in understanding the chemical diversity of coffee secondary metabolites.

Desempenho de frangos de corte alimentados com diferentes níveis de suplementação de vitamina E / Effect of vitamin E supplementation on the performance of broiler chickens

Objetivou-se a avaliação dos efeitos dos níveis de suplementação de vitamina E na ração para frangos de corte sobre o desempenho na fase inicial (um a 21 dias de idade) e de crescimento (21 a 39 dias de idade). Os tratamentos foram definidos pelos níveis de suplementação de vitamina E (DL-α tocoferol). As quantidades adicionadas às rações foram: 10; 30; 50; 75 e 100mg de vitamina E para cada kg de ração. Na fase inicial (experimento I), o menor nível de suplementação de vitamina E mostrou-se superior aos demais, apresentando efeito significativo (P≤0,05) sobre o peso médio aos 21 dias de idade e ganho de peso de um a 21 dias de idade. Na fase de crescimento (experimento II), não houve efeito significativo (P>0,05) dos níveis de suplementação de vitamina E sobre as variáveis estudadas. O nível mínimo de suplementação de vitamina E testado (10mg/kg) é suficiente para atender às exigências dos frangos de corte machos sobre o desempenho.(AU)

This study aimed to evaluate the effect of vitamin E supplementation on the performance of broiler chickens in the initial and growth phase, from one to 21 and 21 to 39 days of age, respectively. Five increasing levels of DL-α tocopherol supplementation (10, 30, 50, 75 and 100mg to each kg of diet) were added to the diet. In the initial phase (experiment I), the lowest vitamin E level was superior by presenting significant effect (P≤0.05) on body weight at 21 days of age and weight gain from one to 21 days old. In the growth phase (experiment II), there was no significant effect (P>0.05) of vitamin E supplementation on the variables studied. The minimum level of vitamin E supplementation tested is sufficient to attend the requirements of male broiler chickens.(AU)

Efeito da protease sobre o coeficiente de metabolizabilidade dos nutrientes em frangos de corte / Effect of protease on the metabolization coefficient of nutrients in broilers

Avaliaram-se os efeitos da protease sobre o coeficiente de metabolizabilidade dos nutrientes em dietas contendo farinha de penas (2,0% na fase inicial e 3,0% na fase de crescimento) para frangos de corte machos, Cobb(r), de um a 32 dias de idade. Foram alojadas 336 aves em gaiolas metálicas equipadas com bandeja para coleta total de excretas. Foi utilizado delineamento inteiramente ao acaso, em arranjo fatorial 2x2 (com e sem adição da enzima protease 0,05% e duas valorizações da matriz nutricional da enzima), sendo sete repetições por tratamento, de 12 aves cada. Observou-se, na fase inicial, maior CMPB (P≤0,05) para as aves que, independentemente da adição de protease, consumiram ração com valorização da matriz nutricional da enzima. Não houve efeito dos tratamentos sobre o CMMS e o CMEE (P>0,05). Na fase de crescimento, houve interação entre os tratamentos para as variáveis CMPB e CMEE. Os frangos alimentados com dieta valorizada sem adição de enzima (controle negativo) apresentaram melhores resultados (P≤0,05) para CMPB e CMEE. Nos tratamentos com enzima, o melhor CMEE (P≤0,05) foi obtido com o tratamento sem valorização da matriz nutricional (over the top). Conclui-se que níveis reduzidos e enzima sem valorização melhoram os coeficientes de metabolizabilidade dos nutrientes na fase adulta de frangos de corte.(AU)

This experiment aimed to evaluate the effects of protease on metabolization coefficient of nutrients, performance and slaughter yield from male Cobb(r) broilers fed diets with feather meal. Two experiments were carried out in a completely randomized design in a 2x2 factorial arrangement - with (0.05%) or without enzyme addition x considering or not the improvement of the nutritional value by protease, with seven replicates per treatment group. In experiment 1 the metabolization coefficient of dry matter (MCDM), crude protein (MCCP) and ether extract (MCEE) in initial (9-12-d-old) and growth (29-32-d-old) phase was evaluated. In the initial phase it was observed that regardless of enzyme addition, broiler chickens fed a diet considering improvement in nutritional value had higher MCCP (P≤0.05). However, MCDM and MCEE were not affected (P>0.05). In the growth phase, there were interactions between treatments for the variables MCCP and MCEE. The birds fed diets considering the improvement in nutritional value without enzyme (negative control) showed better results (P≤0.05) for MCCP and MCEE. In the treatments with enzyme, the best MCEE (P≤0.05) was observed in the treatment not considering the improvement in the nutritional value (over the top). Therefore, considering the improvement in the nutritional value and enzyme without considering the improvement in the nutritional value improves the coefficients of metabolization of nutrients in broiler chickens.(AU)

CoffeebEST: an integrated resource for Coffea spp expressed sequence tags.

Coffee is one of the most important commodities in the world, and its production relies mainly on two species, Coffea arabica and Coffea canephora. Although there are diverse transcriptome datasets available for coffee trees, few research groups have exploited the potential knowledge contained in these data, especially with respect to fruit and seed development. Here, we present a comparative analysis of the transcriptomes of Coffea arabica and Coffea canephora with a focus on fruit development using publicly available expressed sequence tags (ESTs). Most of the fruit and seed EST data has been obtained from C. canephora. Therefore, we performed a fruit EST analysis of the 5 developmental stages of this species (18, 22, 30, 42, and 46 weeks after flowering) comprising 29,009 sequences. We compared C. canephora fruit ESTs to reference unigenes of C. canephora (7710 contigs and 8955 singletons) and C. arabica (15,656 contigs and 16,351 singletons). Additional analyses included functional annotation based on Gene Onthology, as well as an annotation using PlantCyc, a curated plant protein database. The Coffee Bean EST (CoffeebEST) is a public database available at http://bioinfo-02.cp.utfpr.edu.br/. This database represents an additional resource for the coffee scientific community, offering a user-friendly collection of information for non-specialists in coffee molecular biology to support experimental research on comparative and functional genomics.

Construction and characterization of a BAC library from the Coffea arabica genotype Timor Hybrid CIFC 832/2.

Most of the world's coffee production originates from Coffea arabica, an allotetraploid species with low genetic diversity and for which few genomic resources are available. Genomic libraries with large DNA fragment inserts are useful tools for the study of plant genomes, including the production of physical maps, integration studies of physical and genetic maps, genome structure analysis and gene isolation by positional cloning. Here, we report the construction and characterization of a Bacterial Artificial Chromosome (BAC) library from C. arabica Timor Hybrid CIFC 832/2, a parental genotype for several modern coffee cultivars. The BAC library consists of 56,832 clones with an average insert size of 118 kb, which represents a dihaploid genome coverage of five to sixfold. The content of organellar DNA was estimated at 1.04 and 0.5 % for chloroplast and mitochondrial DNA, respectively. The BAC library was screened for the NADPH-dependent mannose-6-phosphate reductase gene (CaM6PR) with markers positioned on four linkage groups of a partial C. arabica genetic map. A mixed approach using PCR and membrane hybridization of BAC pools allowed for the discovery of nine BAC clones with the CaM6PR gene and 53 BAC clones that were anchored to the genetic map with simple sequence repeat markers. This library will be a useful tool for future studies on comparative genomics and the identification of genes and regulatory elements controlling major traits in this economically important crop species.

Gene expression and enzymatic activity of pectin methylesterase during fruit development and ripening in Coffea arabica L.

Coffee quality is directly related to the harvest and post harvest conditions. Non-uniform maturation of coffee fruits, combined with inadequate harvest, negatively affects the final quality of the product. Pectin methylesterase (PME) plays an important role in fruit softening due to the hydrolysis of methylester groups in cell wall pectins. In order to characterize the changes occurring during coffee fruit maturation, the enzymatic activity of PME was measured during different stages of fruit ripening. PME activity progressively increased from the beginning of the ripening process to the cherry fruit stage. In silico analysis of expressed sequence tags of the Brazilian Coffee Genome Project database identified 5 isoforms of PME. We isolated and cloned a cDNA homolog of PME for further characterization. CaPME4 transcription was analyzed in pericarp, perisperm, and endosperm tissues during fruit development and ripening as well as in other plant tissues. Northern blot analysis revealed increased transcription of CaPME4 in the pericarp 300 days after flowering. Low levels of CaPME4 mRNAs were observed in the endosperm 270 days after flowering. Expression of CaPME4 transcripts was strong in the branches and lower in root and flower tissues. We showed that CaPME4 acts specifically during the later stages of fruit ripening and possibly contributes to the softening of coffee fruit, thus playing a significant role in pectin degradation in the fruit pericarp.

Expression patterns of three α-expansin isoforms in Coffea arabica during fruit development.

As a first step towards understanding the physiological role and regulation of the expansin gene (EXP) family in Coffea arabica fruits during growth and maturation, we identified 11 expansin genes, nine belonging to the α-expansin family (EXPA), one EXLA and one EXLB, through in silico analysis of expressed sequence tags (ESTs). Within the α-expansin family, three isoforms were selected for detailed examination based on their high expression in coffee fruits or because they were specifically induced during different fruit developmental stages, according to the EST information. The expression patterns were analysed in different fruit tissues (perisperm, endosperm and pericarp) of C. arabica cv. IAPAR-59 and C. arabica cv. IAPAR-59 Graúdo, the latter being a closely related cultivar with a larger fruit size. Accumulation of CaEXPA1 and CaEXPA3 transcripts was high in the perisperm (tissue responsible for coffee bean size) and in the early stages of pericarp development. Transcripts of CaEXPA2 were detected only in the pericarp during the later stages of fruit maturation and ripening. There was no detectable transcription of the three EXPs analysed in the endosperm. The observed differences in mRNA expression levels of CaEXPA1 and CaEXP3 in the perisperm of IAPAR-59 and IAPAR-59 Graúdo suggest the participation of these two isoforms in the regulation of grain size.

Reference gene selection for real-time quantitative polymerase chain reaction normalization in "Swingle" citrumelo under drought stress.

We describe the first systematic evaluation of reference genes for use in real-time quantitative polymerase chain reaction (qPCR) for water deficit stress studies in the citrus rootstock "Swingle" citrumelo. The expression levels of seven reference genes-cyclophilin (CYP), cathepsin (CtP), actin (ACT), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), elongation factor 1alpha (EF1alpha), beta-tubulin (TUB), and ADP ribosylation factor (ADP)-during drought stress were tested using geNorm and NormFinder programs. Results from four experimental conditions indicated that EF1alpha and ADP were the most stable reference genes. Relative expression levels of Delta1-pyrroline-5-carboxylate synthetase (P5CS) was used for reference gene validation.