Amino acid sequences of proteins from Leptospira serovar pomona

This report describes a partial amino acid sequences from three putative outer envelope proteins from Leptospira serovar pomona. In order to obtain internal fragments for protein sequencing, enzymatic and chemical digestion was performed. The enzyme clostripain was used to digest the proteins 32 and 45 kDa. In situ digestion of 40 kDa molecular weight protein was accomplished using cyanogen bromide. The 32 kDa protein generated two fragments, one of 21 kDa and another of 10 kDa that yielded five residues. A fragment of 24 kDa that yielded nineteen residues of amino acids was obtained from 45 kDa protein. A fragment with a molecular weight of 20 kDa, yielding a twenty amino acids sequence from the 40 kDa protein.

Mapping the molecular characteristics of Brazilian human T-cell lymphotropic virus type 1 Env (gp46) and Pol amino acid sequences for vaccine design

This study was carried out to evaluate the molecular pattern of all available Brazilian human T-cell lymphotropic virus type 1 Env (n = 15) and Pol (n = 43) nucleotide sequences via epitope prediction, physico-chemical analysis, and protein potential sites identification, giving support to the Brazilian AIDS vaccine program. In 12 previously described peptides of the Env sequences we found 12 epitopes, while in 4 peptides of the Pol sequences we found 4 epitopes. The total variation on the amino acid composition was 9 and 17% for human leukocyte antigen (HLA) class I and class II Env epitopes, respectively. After analyzing the Pol sequences, results revealed a total amino acid variation of 0.75% for HLA-I and HLA-II epitopes. In 5 of the 12 Env epitopes the physico-chemical analysis demonstrated that the mutations magnified the antigenicity profile. The potential protein domain analysis of Env sequences showed the loss of a CK-2 phosphorylation site caused by D197N mutation in one epitope, and a N-glycosylation site caused by S246Y and V247I mutations in another epitope. Besides, the analysis of selection pressure have found 8 positive selected sites (w = 9.59) using the codon-based substitution models and maximum-likelihood methods. These studies underscore the importance of this Env region for the virus fitness, for the host immune response and, therefore, for the development of vaccine candidates.

Molecular characterisation of the NSP4 gene of group A human rotavirus G2P[4] strains circulating in São Paulo, Brazil, from 1994 and 2006 to 2010

Group A human rotaviruses (HuRVA) are causative agents of acute gastroenteritis. Six viral structural proteins (VPs) and six nonstructural proteins (NSPs) are produced in RV-infected cells. NSP4 is a diarrhoea-inducing viral enterotoxin and NSP4 gene analysis revealed at least 15 (E1-E15) genotypes. This study analysed the NSP4 genetic diversity of HuRVA G2P[4] strains collected in the state of São Paulo (SP) from 1994 and 2006-2010 using reverse transcription-polymerase chain reaction, sequencing and phylogenetic analysis. Forty (97.6%) G2P[4] strains displayed genotype E2; one strain (2.4%) displayed genotype E1. These results are consistent with the proposed linkage between VP4/VP7 (G2P[4]) and the NSP4 (E2) genotype of HuRVA. NSP4 phylogenetic analysis showed distinct clusters, with grouping of most strains by their genotype and collection year, and most strains from SP were clustered together with strains from other Brazilian states. A deduced amino acid sequence alignment for E2 showed many variations in the C-terminal region, including the VP4-binding domain. Considering the ability of NSP4 to generate host immunity, monitoring NSP4 variations, along with those in the VP4 or VP7 protein, is important for evaluating the circulation and pathogenesis of RV. Finally, the presence of one G2P[4]E1 strain reinforces the idea that new genotype combinations emerge through reassortment and independent segregation.

Preliminary evaluation of acid mine drainage in Minas Gerais State, Brazil

Mining in the State of Minas Gerais-Brazil is one of the activities with the strongest impact on the environment, in spite of its economical importance. Amongst mining activities, acid drainage poses a serious environmental problem due to its widespread practice in gold-extracting areas. It originates from metal-sulfide oxidation, which causes water acidification, increasing the risk of toxic element mobilization and water resource pollution. This research aimed to evaluate the acid drainage problem in Minas Gerais State. The study began with a bibliographic survey at FEAM (Environment Foundation of Minas Gerais State) to identify mining sites where sulfides occur. Substrate samples were collected from these sites to determine AP (acidity potential) and NP (neutralization potential). The AP was evaluated by the procedure of the total sulfide content and by oxygen peroxide oxidation, followed by acidity titration. The NP was evaluated by the calcium carbonate equivalent. Petrographic thin sections were also mounted and described with a special view to sulfides and carbonates. Based on the chemical analysis, the acid-base accounting (ABA) was determined by the difference of AP and NP, and the acid drainage potential obtained by the ABA value and the total volume of material at each site. Results allowed the identification of substrates with potential to generate acid drainage in Minas Gerais state. Altogether these activities represent a potential to produce between 3.1 to 10.4 billions of m³ of water at pH 2 or 31.4 to 103.7 billions of m³ of water at pH 3. This, in turn, would imply in costs of US$ 7.8 to 25.9 millions to neutralize the acidity with commercial limestone. These figures are probably underestimated because some mines were not surveyed, whereas, in other cases, surface samples may not represent reality. A more reliable state-wide evaluation of the acid drainage potential would require further studies, including a larger number of samples. Such investigations should consider other mining operations beyond the scope of this study as well as the kinetics of the acid generation by simulated weathering procedures.

Total lipid and fatty acid accumulation during basidiospore formation in the ectomycorrhizal fungus Pisolithus sp.

The basidiospores of Pisolithus sp. contain large amounts of lipids, indicating provision for future germination in the host rhizosphere. However, the accumulation, composition, and mobilization of lipids during formation and germination of these spores are largely unknown. In this study, lipid storage and fatty acid composition during basidiosporogenesis were analyzed in fresh basidiocarps using bright-field microscopy and gas chromatography. Abundant lipid bodies are found in the hyphae, basidia, and basidiospores of fungal basidiocarps. This evidences a considerable C transport in the basidiocarp to meet the C demand during basidiospore formation. Fatty acid composition analysis revealed the presence of 24 compounds with chains of 9 to 18 C atoms, either saturated or insaturated, with one or two insaturations. The fatty acid composition and content varied according to the developmental stage of the peridioles. In free basidiospores, the predominant compounds were 16:0, 16:1w5c, 18:1w9c, and 18:2w6,9c/18:0ante, at concentrations of 76, 46, 192, and 51 µg g-1 dry matter, respectively. Our results indicate that oleic acid is the major constituent of lipid reserves in Pisolithus sp. basidiospores. Further studies are being conducted to determine the factors that induce lipid mobilization during spore germination.

Liming influence on soil chemical properties, nutritional status and yield of alfalfa grown in acid soil

Alfalfa is an important forage crop with high nutritive value, although highly susceptible to soil acidity. Liming is one of the most efficient and prevailing practices to correct soil acidity and improve alfalfa yield. The objective of this study was to evaluate response to liming of alfalfa grown in a greenhouse on a Typic Quartzipsamment soil. The treatments consisted of four lime rates (0, 3.8, 6.6 and 10.3 Mg ha-1) and two cuts. Alfalfa dry matter increased quadratically with increasing lime rates. In general, dry matter yield was maximized by a lime rate of 8.0 Mg ha-1. Except for the control, the dry matter nutrient contents in the treatments were adequate. The positive linear correlation between root and nodule dry matter with lime rates indicated improvement of these plant traits with decreasing soil acidity. The soil acidity indices pH, base saturation, Ca2+ concentration, Mg2+ concentration, and H + Al were relevant factors in the assessment of alfalfa yield. The magnitude of influence of these soil acidity indices on yield as determined by the coefficient of determination (R²) varied and decreased in the order: base saturation, H + Al, pH, Ca and Mg concentrations. Optimum values of selected soil chemical properties were defined for maximum shoot dry matter; these values can serve as a guideline for alfalfa liming to improve the yield of this forage on acid soils.

Total fatty acid composition in the characterization and identification of orchid mycorrhizal fungi Epulorhiza spp.

Rhizoctonia-like fungi are the main mycorrhizal fungi in orchid roots. Morphological characterization and analysis of conserved sequences of genomic DNA are frequently employed in the identification and study of fungi diversity. However, phytopathogenic Rhizoctonia-like fungi have been reliably and accurately characterized and identified through the examination of the fatty acid composition. To evaluate the efficacy of fatty acid composition in characterizing and identifying Rhizoctonia-like mycorrhizal fungi in orchids, three Epulorhiza spp. mycorrhizal fungi from Epidendrum secundum, two unidentified fungi isolated from Epidendrum denticulatum, and a phytopathogenic fungus, Ceratorhiza sp. AGC, were grouped based on the profile of their fatty acids, which was assessed by the Euclidian and Mahalanobis distances and the UPGMA method. Dendrograms distinguished the phytopathogenical isolate of Ceratorhiza sp. AGC from the mycorrhizal fungi studied. The symbionts of E. secundum were grouped into two clades, one containing Epulorhiza sp.1 isolates and the other the Epulorhiza sp.2 isolate. The similarity between the symbionts of E. denticulatum and Epulorhiza spp. fungi suggests that symbionts found in E. denticulatum may be identified as Epulorhiza. These results were corroborated by the analysis of the rDNA ITS region. The dendrogram constructed based on the Mahalanobis distance differentiated the clades most clearly. Fatty acid composition analysis proved to be a useful tool for characterizing and identifying Rhizoctonia-like mycorrhizal fungi.

Application of alkaline waste from pulp industry to acid soil with pine

In Brazil extensive areas are covered with pine forests, planted for pulp and paper production. This industry generates solid alkaline waste, such as dregs. The application of this dregs to forest soils is an alternative for soil acidity correction and plant nutrient supply, as well as a solution for its proper disposal. The purpose of this study was to compare the residual effect of surface application of dregs and dolomitic lime on (a) changes in the physical and chemical properties of an acidic soil and (b) pine tree development. The experiment was carried out in 2004 in Bocaina do Sul, Santa Catarina, consisting of the application of increasing dreg and lime rates to a Pinus taeda L. production area, on a Humic Cambisol, in a randomized block design with four replications and 10 x 10 m plots. The treatments consisted of levels of soil acidity amendments corresponding to the recommendations by the SMP method to reach pH 5.5 in the 0-20 cm layer, as follows: no soil amendment; dregs at 5.08 (1/4 SMP), 10.15 (1/2 SMP) and 20.3 Mg ha-1 (1 SMP); and lime at 8.35 (1/2 SMP) and 16.7 Mg ha-1 (1 SMP). Soil layers were sampled in 2010 for analyses of soil chemical and physical properties. The diameter at breast height of the 6.5 year old pine trees was also evaluated. Surface application of dregs improved soil chemical fertility by reducing acidity and increasing base saturation, similar to liming, especially in surface layers. Dregs, comparable to lime, reduced the degree of clay flocculation, but did not affect the soil physical quality. There was no effect of the amendments on increase in pine tree diameter. Thus, the alternative to raise the pH in forest soils to 5.5 with dregs is promising for the forestry sector with a view to dispose of the waste and increase soil fertility.

Organic acid adsorption and mineralization in oxisols with different textures

Organic acids play an important role in the nutritional conditions of plants. Their relevance is related to their formation dynamics, mineralization rate and adsorption by soil colloids. This study was carried out to evaluate the dynamics of mineralization and adsorption of organic acid (acetic acid - AA, citric acid - CA and humic acid - HA) applied to the soil. Samples of two Oxisols were used: Rhodic Haplustox (LV) and Typic Haplustox (LVA). The mineralization experiment was arranged in a 2 x 3 x 5 factorial design, based on the factors: two soils (LV and LVA) x three organic acid (OA) types (AA, CA and HA) x five OA rates (0, 1, 2, 4, and 8 mmol dm-3). Organic carbon mineralization in samples was measured by the C-CO2 efflux, produced by the microbial activity, in a 30-day (measurements after 4, 8, 12, 21, and 30 days) and in a 4-day experiment (measured after 24, 48, 72 and 96 h). Organic acid adsorption was tested in a 2 x 2 x 5 x 4 factorial design, with the factors and levels: two Oxisols; two organic acids (AA and CA); five OA rates (0, 1, 2, 4, and 8 mmol dm-3) and four adsorption periods (6, 24, 48, and 72 h). The C-CO2 production of soil treated with CA was highest. In the adsorption experiment, the affinity of CA to soil adsorption sites was greatest. The adsorption of organic acids to soils may be an important mechanism by which bioavailability and thus mineralization capacity by microbial activity are reduced.

Characterization of Ornamental Rock Residue and Potassium Liberation Via Organic Acid Application

ABSTRACT Organic acids present in organic matter and, or, exudates by microorganisms and plants can increase the liberation of potassium present in minerals. The objective of this study was to characterize the residue from ornamental rocks and evaluate the release of K from these residues after the application of organic acids. The experiment was conducted under laboratory conditions and followed a 2 × 3 × 5 factorial design with three replicates. The studied factors were: two organic acids (citric acid and malic acid), three ornamental rock residues (R1, R2 and R3) and five organic acid rates (0, 5, 10, 20 and 40 mmol L-1). After agitation, K concentrations were determined in the equilibrium solution. Successive extractions were performed (1, 5, 10, 15, 30 and 60 days after the start of the experiment). The organic acids used (citric and malic) promoted the release of up to 4.86 and 4.34 % of the total K contained in the residue, respectively, reinforcing the role of organic acids in the weathering of minerals and in providing K to the soil. The K quantities were, on average, 6.1 % higher when extracted with citric acid compared to malic acid.

Matriconditioning integrated with gibberellic acid to hasten seed germination and improve stand establishment of pepper and tomato

Emergence and stand establishment of tomato (Lycopersicon lycopersicum (L.) (Karsten ex Farw) and pepper (Capsicum annus L.) seeds are often slow and erratic, particularly under stress conditions. Field emergence trials sometimes have not responded to priming in pepper. This study examined the combined effects of matriconditioning and gibberellin application on the germination and stand establishment of pepper and tomato seeds. Pepper and tomato seeds were conditioned with a solid carrier, Micro Cel E, in the presence of gibberellic acid (GA) for 1, 2 , 3 and 4 days at 15 and 25ºC. The results showed that, in all cases, even under stress conditions, the combination of matriconditioning with GA was effective in improving germination and emergence of pepper and tomato. The germination time was, in average, reduced by 2 to 3 days by primed seeds. Thus, matriconditioning, during which germination is suspended, provides an unique means to rapidly and efficiently digest the endosperm by GA-induced enzymes and reduce the mechanical restraints of endosperm thus providing energy to start and sustain embryo growth.

Dietary effects on muscle fatty acid composition of finished heifers

The effects of diet on Longissimus muscle fatty acid composition was determined using 24 crossbred heifers of Simmental vs. Nelore and Limousin vs. Nelore. The experimental diets were: 1) corn and yeast (CY); 2) corn, cottonseed meal + meat and bones meal (CMB); 3) cassava hull and yeast (CHY); 4) cassava hull, cottonseed meal + meat and bones meal (CHMB). Feeding CHMB diets resulted in lower lipid and higher cholesterol contents (P<0.05) for both crosses. Most of the identified fatty acids were monounsaturated, and the highest percentage was found to oleic acid (C18:1w9), with values ranging from 32.54 to 46.42%. Among the saturated fatty acids the palmitic acid (C16:0) showed the highest percentage, with its contents ranging between 19.40 and 32.44%. The highest polyunsaturated/saturated fatty acid ratio was of 0.30, and the lowest was of 0.08. Feeding CY diets resulted in lower cholesterol and higher polyunsaturated fatty acid contents of the Longissimus muscle.

Evaluation of acid digestion techniques to estimate chromium contents in cattle feces

The objective of this work was to evaluate the accuracy of digestion techniques using nitric and perchloric acid at the ratios of 2:1, 3:1, and 4:1 v v-1, in one- or two-step digestion, to estimate chromium contents in cattle feces, using sodium molybdate as a catalyst. Fecal standards containing known chromium contents (0, 2, 4, 6, 8, and 10 g kg-1) were produced from feces of five animals. The chromium content in cattle feces is accurately estimated using digestion techniques based on nitric and perchloric acids, at a 3:1 v v-1 ratio, in one-step digestion, with sodium molybdate as a catalyst.

Indol-butyric acid levels on cashew cloning by air-layering process

A study was conducted to determine the possibility of cashew (Anacardium occidentale) cloning by air-layering and influence of IBA (indol-butyric acid) on this process. It was adopted a completely randomized design with 4 treatments, 10 air layers each and 4 replications, reaching 160 air layers. The IBA levels on the treatments were, as follow: 0, 1000, 3000 and 5000 mg.kg-1. It was evaluated: survival, callus and rooting percentage, average number and length of roots. The highest survival rate (67.5%) was registered with no growth regulator and IBA at 1000 mg.kg-1, while the best rooting percentage (82%) referred to 1000 mg.kg-1. In spite of average number and length of roots, the highest results were observed with IBA at 5000 mg.kg-1. IBA concentrations had no influence on cashew air-layering formation.

Soil nutrient availability and its impact on fruit quality of Tahiti acid lime

The Tahiti acid lime in Brazil is mostly grown in the São Paulo State. The value of this crop production ranks among the ten most important fruits in the country. The Brazilian exports of Tahiti limes have increased in the last years with a corresponding increased demand for superior quality of fresh fruits, which is affected by mineral nutrients. Therefore, this study evaluated nutrient soil availability and its influence on nutritional status of trees based on the determination of leaf and fruit nutrient concentrations, fruit characteristics, and post harvest quality. Eleven commercial groves with trees older than 4-yr and differently managed were studied. Plots with six trees in each grove were sampled for soil (0-20 cm depth layer), leaf and fruit analyses with three replicates. Correlation coefficients were pair wised established for all variables. The results showed that N leaf concentration was well correlated with green color of fruit peel as measured by a color index (r = -0.71**), and which was optimum with Leaf-N around 22 g kg-1. Leaf-Ca was inversely correlated with fruit water loss after 14-day interval from harvest (r = -0.54*) demonstrating that Ca plays an important role in Tahiti fruit shelf-life. Data also suggested that increased fruit K concentration correlated with increased fruit water losses during storage (r >0.58*).