Salmonella enterica serovar typhimurium colonizing the lumen of the chicken intestine grows slowly and upregulates a unique set of virulence and metabolism genes

The pattern of global gene expression in Salmonella enterica serovar Typhimurium bacteria harvested from the chicken intestinal lumen (cecum) was compared with that of a late-log-phase LB broth culture using a whole-genome microarray. Levels of transcription, translation, and cell division in vivo were lower than those in vitro. S. Typhimurium appeared to be using carbon sources, such as propionate, 1,2-propanediol, and ethanolamine, in addition to melibiose and ascorbate, the latter possibly transformed to D-xylulose. Amino acid starvation appeared to be a factor during colonization. Bacteria in the lumen were non- or weakly motile and nonchemotactic but showed upregulation of a number of fimbrial and Salmonella pathogenicity island 3 (SPI-3) and 5 genes, suggesting a close physical association with the host during colonization. S. Typhimurium bacteria harvested from the cecal mucosa showed an expression profile similar to that of bacteria from the intestinal lumen, except that levels of transcription, translation, and cell division were higher and glucose may also have been used as a carbon source. © 2011, American Society for Microbiology.

Levels of digestible threonine for male broilers from 1 to 7 days of age

The influence of threonine has been studied in broilers; however, information concerning the phase from 1 to 7 d is lacking. In this study, we evaluated the effects on performance and body composition and the relationship with other amino acids with respect to digestible lysine and total glycine + glycine. Research on the initial 7-d phase of broiler growth is still poorly studied, even though it is important, especially in nutrition. It was found that threonine had a great influence on the broilers at that phase and nutritional recommendations need constant evaluation. In conclusion, the digestible threonine: digestible lysine ratio and digestible threonine: total glycine + serine ratio recommended for better performance and body composition are 0.68 and 0.40, respectively. The levels of digestible threonine and total glycine + serine that provide the best results are 0.864 and 2.192%, respectively.

Increased pancreatic islet mass is accompanied by activation of the insulin receptor substrate-2/serine-threonine kinase pathway and augmented cyclin D(2) protein levels in insulin-resistant rats

It is well known that glucocorticoids induce peripheral insulin resistance in rodents and humans. Here, we investigated the structural and ultrastructural modifications, as well as the proteins involved in beta-cell function and proliferation, in islets from insulin-resistant rats. Adult male Wistar rats were made insulin resistant by daily administration of dexamethasone (DEX; 1mg/kg, i.p.) for five consecutive days, whilst control (CTL) rats received saline alone. Structure analyses showed a marked hypertrophy of DEX islets with an increase of 1.7-fold in islet mass and of 1.6-fold in islet density compared with CTL islets (P < 0.05). Ultrastructural evaluation of islets revealed an increased amount of secreting organelles, such as endoplasmic reticulum and Golgi apparatus in DEX islets. Mitotic figures were observed in DEX islets at structural and ultrastructural levels. Beta-cell proliferation, evaluated at the immunohistochemical level using anti-PCNA (proliferating cell nuclear antigen), showed an increase in pancreatic beta-cell proliferation of 6.4-fold in DEX islets compared with CTL islets (P < 0.0001). Increases in insulin receptor substrate-2 (IRS-2), phosphorylated-serine-threonine kinase AKT (p-AKT), cyclin D(2) and a decrease in retinoblastoma protein (pRb) levels were observed in DEX islets compared with CTL islets (P < 0.05). Therefore, during the development of insulin resistance, the endocrine pancreas adapts itself increasing beta-cell mass and proliferation, resulting in an amelioration of the functions. The potential mechanisms that underlie these events involve the activation of the IRS-2/AKT pathway and activation of the cell cycle, mediated by cyclin D(2). These adaptations permit the maintenance of glycaemia at near-physiological ranges.

Structural studies of prephenate dehydratase from Mycobacterium tuberculosis H37Rv by SAXS, ultracentrifugation, and computational analysis

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Crystallization and preliminary X-ray diffraction analysis of prephenate dehydratase from Mycobacterium tuberculosis H37Rv

Tuberculosis remains the leading cause of mortality arising from a bacterial pathogen ( Mycobacterium tuberculosis). There is an urgent need for the development of new antimycobacterial agents. The aromatic amino-acid pathway is essential for the survival of this pathogen and represents a target for structure-based drug design. Accordingly, the M. tuberculosis prephenate dehydratase has been cloned, expressed, purified and crystallized by the hanging-drop vapour-diffusion method using PEG 400 as a precipitant. The crystal belongs to the orthorhombic space group I222 or I2(1)2(1)2(1), with unit-cell parameters a = 98.26, b = 133.22, c = 225.01 angstrom, and contains four molecules in the asymmetric unit. A complete data set was collected to 3.2 angstrom resolution using a synchrotron-radiation source.

Maintenance requirements for methionine and cysteine, and threonine for poultry

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

Protective effect of mannan oligosaccharides against early colonization by Salmonella Enteritidis in chicks is improved by higher dietary threonine levels

Aims: To evaluate mannan oligosaccharide (MOS) and threonine effects on performance, small intestine morphology and Salmonella spp. counts in Salmonella Enteritidis-challenged birds. Methods and Results: One-day-old chicks (1d) were distributed into five treatments: nonchallenged animals fed basal diet (RB-0), animals fed basal diet and infected with Salmonella Enteritidis (RB-I), animals fed high level of threonine and infected (HT-I), birds fed basal diet with MOS and infected (MOS-I), birds fed high level of threonine and MOS and infected (HT+MOS-I). Birds were inoculated at 2d with Salmonella Enteritidis, except RB-0 birds. Chicks fed higher dietary threonine and MOS showed performance similar to RB-0 and intestinal morphology recovery at 8 dpi. Salmonella counts and the number of Salmonella-positive animals were lower in HT+MOS-I compared with other challenged groups. Conclusion: Mannan oligosaccharides and threonine act synergistically, resulting in improved intestinal environment and recovery after Salmonella inoculation. Significance and Impact of the Study: Nutritional approaches may be useful to prevent Salmonella infection in the first week and putative carcass contamination at slaughter. This is the first report on the possible synergistic effect of mannan oligosaccharides and threonine, and further studies should be performed including performance, microbiota evaluation, composition of intestinal mucins and immune assessment. © 2012 The Society for Applied Microbiology.

The optimal lysine and threonine intake for Cobb broiler breeder hens using Reading model

This study aimed to determine the optimal intake of lysine and threonine for broiler breeder hens. Two experiments were conducted to evaluate the responses of birds to digestible lysine (Lys) and threonine (Thr). Eight treatments were assessed in both experiments, with six replicates of eight birds in the Lys experiment and ten birds in the Thr experiment. The dietary levels of Lys and Thr were obtained by a dilution technique. The experimental period was ten weeks for each amino acid studied, which included six weeks of adaptation and four weeks of data collection. The amino acid intake, egg mass and body weight were adjusted using a Reading model. Based on the model coefficients, the cost of the synthetic amino acids sources and the price of fertile eggs determined the intake of each amino acid to maximize. The minimum intake of Lys and Thr reduced egg production by 40 and 30%, respectively, the weight of the eggs decreased by 12 and 9% with the same intake of Lys and Thr, respectively. The models generated by predicting Lys and Thr intake were as follows: Lys=11 x E+31 x W and Thr=9.5 x E+32 x W, where E=egg mass, g/bird per day, and W=body weight, kg/bird. Based on the models, 3 kg birds with an egg mass production of 50 g/day require 643 mg/bird per day of Lys and 569 mg/bird per day of Thr. The optimum economic intake was calculated at 954 and 834 mg/bird per day for Lys and Thr, respectively, reflecting a dietary concentration of 0.636% Lys and 0.556% Thr for a feed intake of 150 g/bird per day. (C) 2015 Elsevier B.V. All rights reserved.

The response to dietary threonine in laying-type pullets during growth

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

Pool of Resistance Mechanisms to Glyphosate in Digitaria insularis

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Diferentes critérios de formulação de rações para frangos de corte no período de 1 a 21 dias de idade

Dois experimentos foram realizados com o objetivo de avaliar diferentes critérios de formulação de rações, baseados em aminoácidos totais e digestíveis, para frangos de corte no período de 1 a 21 dias de idade. As dietas foram formuladas para atender às exigências de metionina, metionina+cistina, lisina e treonina. No primeiro experimento, foram utilizados 800 pintos machos, Hubbard, distribuídos em delineamento inteiramente casualizado com 4 critérios de formulação, para atender às exigências em aminoácidos totais - NRC (1994), Rostagno et al. (1992), Degussa (1997) e AEC (1993), com 4 repetições de 50 aves. No segundo experimento, foram utilizados 600 pintos machos, Cobb, distribuídos em delineamento inteiramente casualizado com 3 critérios de formulação, para atender às exigências em aminoácidos digestíveis - Rostagno et al. (2000), Baker & Han (1994) e Degussa (1997), com 4 repetições de 50 aves. Foram avaliados ganho de peso, consumo de ração e conversão alimentar. Os resultados obtidos demonstraram não haver diferença no desempenho das aves ao serem alimentadas com diferentes critérios de formulação baseados em aminoácidos totais. Ao formular dietas com aminoácidos digestíveis, o padrão estabelecido por Baker & Han (1994) resultaram em melhor desempenho das aves no período avaliado.

Avaliação do Desempenho e Excreção de Cálcio em Duas Linhagens de Frangos de Corte, na Fase Inicial, Alimentados com Diferentes Níveis de Aminoácidos e de Cálcio

Dois experimentos foram realizados com o objetivo de avaliar os efeitos de diferentes níveis de aminoácidos e de cálcio sobre o desempenho e excreção de cálcio de duas linhagens de frangos de corte (Cobb e Avian Farms) na fase inicial de criação. Foram avaliados os dados de desempenho (consumo de ração, ganho de peso e conversão alimentar) e excreção de cálcio. Foram utilizadas 540 aves, em cada experimento, distribuídas em delineamento inteiramente casualizado, em esquema fatorial 3x2, ou seja, 3 perfis de aminoácidos (metionina, metionina+cistina, lisina e treonina - 100%, 125% e 150% dos níveis do NRC,1994) e 2 níveis de cálcio (75% e 100% dos níveis do NRC,1994), com 30 aves por repetição. Analisou-se o cálcio excretado das aves, e para tanto, utilizou-se 72 aves, em cada experimento, alojadas em gaiolas, seguindo o mesmo delineamento descrito anteriormente. Não houve interação entre os tratamentos estudados. Para a linhagem Avian Farms, o ganho de peso diminuiu com a elevação dos níveis de aminoácidos e ocorreu menor excreção de cálcio com menor nível de cálcio da dieta. A linhagem Cobb não foi influenciada pelos tratamentos em nenhum dos parâmetros estudados.

Desempenho, rendimento de carcaça e excreção de cálcio de frangos de corte alimentados com diferentes níveis de aminoácidos e cálcio no período de 22 a 42 dias de idade

Um experimento foi conduzido com o objetivo de avaliar diferentes níveis de aminoácidos e de cálcio para frangos de corte no período de 22 a 42 dias de idade. Avaliou-se o desempenho, o rendimento de carcaça e a excreção de cálcio. Foram utilizadas 540 aves, machos, da marca comercial Cobb, distribuídas em delineamento inteiramente casualizado, em esquema fatorial 3x2, ou seja, 3 níveis de aminoácidos (metionina, metionina+cistina, lisina e treonina - 100%, 125% e 150% NRC,1994) e 2 níveis de cálcio (75% e 100% NRC,1994), com 3 repetições de 30 aves cada. Os aminoácidos estudados foram metionina, metionina+cistina, lisina e treonina. O desempenho das aves foi avaliado pelo peso vivo, ganho de peso, consumo de ração e conversão alimentar. O rendimento de carcaça e a percentagem de cada parte foram avaliados de 2 aves de cada repetição, escolhidas ao acaso, perfazendo um total de 6 animais por tratamento. O rendimento de carcaça foi realizado após o abate levando-se em consideração a percentagem de peito, pernas (coxa+sobrecoxa), asas, dorso, cabeça+pescoço e pés em relação à carcaça eviscerada. Além disso, analisou-se o cálcio excretado pelas aves. Para tanto, utilizou-se 72 aves, alojadas em gaiolas, seguindo o mesmo delineamento descrito anteriormente. Não houve interação entre os fatores estudados para as características avaliadas. As que consumiram dietas com 125% e 150% de aminoácidos apresentaram melhor conversão alimentar. O rendimento de peito foi afetado pelos níveis de cálcio.

Genomics-based design of defined growth media for the plant pathogen Xylella fastidiosa

Based on the genetic analysis of the phytopathogen Xylella fastidiosa genome, five media with defined composition were developed and the growth abilities of this fastidious prokaryote were evaluated in liquid media and on solid plates. All media had a common salt composition and included the same amounts of glucose and vitamins but differed in their amino acid content. XDM1 medium contained amino acids threonine, serine, glycine, alanine, aspartic acid and glutamic acid, for which complete degradation pathways occur in X fastidiosa; XDM2 included serine and methionine, amino acids for which biosynthetic enzymes are absent, plus asparagine and glutamine, which are abundant in the xylem sap; XDM3 had the same composition as XDM2 but with asparagine replaced by aspartic acid due to the presence of complete degradation pathway for aspartic acid; XDM4 was a minimal medium with glutamine as a sole nitrogen source; XDM5 had the same composition as XDM4, plus methionine. The liquid and solidified XDM2 and XDM3 media were the most effective for the growth of X. fastidiosa. This work opens the opportunity for the in silico design of bacterial defined media once their genome is sequenced. (C) 2002 Federation of European Microbiological Societies. Published by Elsevier B.V. B.V. All rights reserved.