Purification and biochemical characterization of four iron superoxide dismutases in Trypanosoma cruzi

Four superoxide dismutase (SOD) activities (SOD I, II, III, and IV) have been characterized in the epimastigote form of Trypanosoma cruzi. The total extract was subjected to two successive ammonium sulphate additions between 35 and 85%, and the resulting fraction was purified using two continuous chromatography processes (ion exchange and filtration). Enzymes were insensitive to cyanide but sensitive to hydrogen peroxide, properties characteristic of iron-containing SODs. The molecular masses of the different SODs were 20 kDa (SOD I), 60 kDa (SOD II), 50 kDa (SOD III) and 25 kDa (SOD IV), whereas the isoelectric points were 6.9, 6.8, 5.2 and 3.8, respectively. Subcellular location and digitonin experiments have shown that these SODs are mainly cytosolic, with small amounts in the low-mass organelles (SOD II and SOD I) and the mitochondrion (SOD III), where these enzymes play an important role in minimizing oxidative damage.

Production of TNF-α, nitric oxide and hydrogen peroxide by macrophages from mice with paracoccidioidomycosis that were fed a linseed oil-enriched diet

Omega-3 polyunsaturated fatty acids (n-3 PUFA) can modulate the immune system and their primary effect is on macrophage function. Paracoccidioidomycosis (PCM) is an endemic systemic mycosis in Latin America that is caused by the dimorphic fungus Paracoccidioides brasiliensis (Pb). Macrophages are the main defence against this pathogen and have microbicidal activity that is dependent on interferon-Γ and tumour necrosis factor (TNF)-α. These cytokines stimulate the synthesis of nitric oxide (NO) and hydrogen peroxide (H2O2), leading to the death of the fungus. To study the effect of n-3 PUFA on the host immune response during experimental PCM, macrophages that were obtained from animals infected with Pb18 and fed a diet enriched by linseed (LIN) oil were cultured and challenged with the fungus in vitro. The macrophage function was analysed based on the concentrations of TNF-α, NO and H2O2. LIN oil seems to influence the production of TNF-α during the development of disease. A diet enriched with LIN oil influences the microbicidal activity of the macrophages by inducing the production of cytokines and metabolites such as NO and H2O2, predominantly in the chronic phase of infection.

The level of ascorbate peroxidase is enhanced in benznidazole-resistant populations of Trypanosoma cruzi and its expression is modulated by stress generated by hydrogen peroxide

Ascorbate peroxidases (APX) are class I heme-containing enzymes that convert hydrogen peroxide into water molecules. The gene encoding APX has been characterized in 11 strains of Trypanosoma cruzi that are sensitive or resistant to benznidazole (BZ). Bioinformatic analysis revealed the presence of two complete copies of the T. cruzi APX (TcAPX) gene in the genome of the parasite, while karyotype analysis showed that the gene was present in the 2.000-kb chromosome of all of the strains analyzed. The sequence of TcAPX exhibited greater levels of similarity to those of orthologous enzymes from Leishmania spp than to APXs from the higher plant Arabidopsis thaliana. Northern blot and real-time reverse transcriptase polymerase chain reaction (RT-PCR) analyses revealed no significant differences in TcAPX mRNA levels between the T. cruzi strains analyzed. On the other hand, Western blots showed that the expression levels of TcAPX protein were, respectively, two and three-fold higher in T. cruzi populations with in vitro induced (17 LER) and in vivo selected (BZR) resistance to BZ, in comparison with their corresponding susceptible counterparts. Moreover, the two BZ-resistant populations exhibited higher tolerances to exogenous hydrogen peroxide than their susceptible counterparts and showed TcAPX levels that increased in a dose-dependent manner following exposure to 100 and 200 µM hydrogen peroxide.

Disinfection with sodium hypochlorite in hospital environmental surfaces in the reduction of contamination and infection prevention: a systematic review

OBJECTIVETo search for evidence of the efficiency of sodium hypochlorite on environmental surfaces in reducing contamination and prevention of healthcare-associated infection HAIs.METHODSystematic review in accordance with the Cochrane Collaboration.RESULTSWe analyzed 14 studies, all controlled trials, published between 1989-2013. Most studies resulted in inhibition of microorganism growth. Some decreased infection, microorganism resistance and colonization, loss of efficiency in the presence of dirty and surface-dried viruses.CONCLUSIONThe hypochlorite is an effective disinfectant, however, the issue of the direct relation with the reduction of HAIs remains. The absence of control for confounding variables in the analyzed studies made the meta-analysis performance inadequate. The evaluation of internal validity using CONSORT and TREND was not possible because its contents were not appropriate to laboratory and microbiological studies. As a result, there is an urgent need for developing specific protocol for evaluating such studies.

Chemical changes in argisols under irrigated grape production in the central São Francisco river valley, Brazil

This study compares the chemical composition of the solution and exchange complex of soil in a 3-year-old irrigated vineyard (Vitis vinifera L., Red Globe cultivar) with that of adjacent clearing in the native hyperxerophyllic 'caatinga' vegetation. The soils are classified as Plinthic Eutrophic Red-Yellow Argisol; according to Soil Taxonomy they are isohyperthermic Plinthustalfs. Detailed physiographic characterization revealed an impermeable gravel and cobble covering the crystalline rocks; the relief of this layer was more undulating than the level surface. Significant higher concentrations of extractable Na, K, Mg and Ca were observed within the vineyard. Lower soil acidity, higher Ca/Mg ratios, as well as lower sodium adsorption and Na/K ratios reflected additions of dolomitic lime, superphosphate and K-bearing fertilizers. As the water of the São Francisco River is of good quality for irrigation (C1S1), the increases in Na were primarily attributed to capillary rise from the saline groundwater table. None of the soil in the study area was found to be sodic. About 62% of the vineyard had an Ap horizon with salinity levels above 1.5dSm-1 (considered detrimental for grape production); according to average values for this horizon, a potential 13% reduction in grape production was predicted. Differences in chemical composition in function of distance to the collector canals were observed in the clearing, but not in the vineyard. The influence of differences in the elevations of the surface and impermeable layers, as well as pediment thickness, was generally weaker under irrigation. Under irrigation, soil moisture was greater in points of convergent surface waterflow; the effect of surface curvature on chemical properties, though less consistent, was also stronger in the vineyard.

Aluminum hydroxy-interlayered minerals and chemical properties of a subtropical Brazilian Oxisol under no-tillage and conventional tillage

No tillage systems significantly influence the soil system, but knowledge about the effects on the mineralogy of tropical and subtropical soils is limited. This study evaluated the long-term effects (26 years) of no-tillage (NT) on aluminum hydroxy-interlayered minerals of a subtropical Oxisol in Southern Brazil (Guarapuava, PR), compared to the same soil under conventional tillage (CT). The clay fraction (< 2 µm) in soil samples of the surface horizons of a field experiment under both management systems was analyzed by X-ray diffraction (XRD) to identify and characterize Al hydroxy-interlayered minerals before and after treatment with sodium citrate to remove intra-layer material. Soil liquid (solution) and solid phases were also characterized. The contents of total organic C, exchangeable cations, P, and the values of extractable acidity and cation exchange capacity as well as electrical conductivity and levels of dissolved organic C, basic cations, aluminum, Si, and sulfur in the soil solution were higher in the NT soil. Under both soil management systems, more than 90 % of the total soluble Al was complexed with organic compounds, with similar Al activity. No significant changes were detected by 2:1 clay mineral XRD analyses in terms of extension or intercalation of Al-hydroxy-polymers in the no-tilled in comparison to the conventionally tilled soil. In both soil management systems, Al and Si activities in the soil solution indicated thermodynamic stability of 2:1 clay minerals with partially occupied by hydroxy-Al, suggesting deceleration in the intercalation process and a tendency of transforming clay minerals from extensive into partial intercalation.

Chemical dispersants and pre-treatments to determine clay in soils with different mineralogy

Knowledge of the soil physical properties, including the clay content, is of utmost importance for agriculture. The behavior of apparently similar soils can differ in intrinsic characteristics determined by different formation processes and nature of the parent material. The purpose of this study was to assess the efficacy of separate or combined pre-treatments, dispersion methods and chemical dispersant agents to determine clay in some soil classes, selected according to their mineralogy. Two Brazilian Oxisols, two Alfisols and one Mollisol with contrasting mineralogy were selected. Different treatments were applied: chemical substances as dispersants (lithium hydroxide, sodium hydroxide, and hexametaphosphate); pre-treatment with dithionite, ammonium oxalate, and hydrogen peroxide to eliminate organic matter; and coarse sand as abrasive and ultrasound, to test their mechanical action. The conclusion was drawn that different treatments must be applied to determine clay, in view of the soil mineralogy. Lithium hydroxide was not efficient to disperse low-CEC electropositive soils and very efficient in dispersing high-CEC electronegative soils. The use of coarse sand as an abrasive increased the clay content of all soils and in all treatments in which dispersion occurred, with or without the use of chemical dispersants. The efficiency of coarse sand is not the same for all soil classes.

Characterization and nutrient release from silicate rocks and influence on chemical changes in soil

The expansion of Brazilian agriculture has led to a heavy dependence on imported fertilizers to ensure the supply of the growing food demand. This fact has contributed to a growing interest in alternative nutrient sources, such as ground silicate rocks. It is necessary, however, to know the potential of nutrient release and changes these materials can cause in soils. The purpose of this study was to characterize six silicate rocks and evaluate their effects on the chemical properties of treated soil, assessed by chemical extractants after greenhouse incubation. The experimental design consisted of completely randomized plots, in a 3 x 6 factorial scheme, with four replications. The factors were potassium levels (0-control: without silicate rock application; 200; 400; 600 kg ha-1 of K2O), supplied as six silicate rock types (breccia, biotite schist, ultramafic rock, phlogopite schist and two types of mining waste). The chemical, physical and mineralogical properties of the alternative rock fertilizers were characterized. Treatments were applied to a dystrophic Red-Yellow Oxisol (Ferralsol), which was incubated for 100 days, at 70 % (w/w) moisture in 3.7 kg/pots. The soil was evaluated for pH; calcium and magnesium were extracted with KCl 1 mol L-1; potassium, phosphorus and sodium by Mehlich 1; nickel, copper and zinc with DTPA; and the saturation of the cation exchange capacity was calculated for aluminum, calcium, magnesium, potassium, and sodium, and overall base saturation. The alternative fertilizers affected soil chemical properties. Ultramafic rock and Chapada mining byproduct (CMB) were the silicate rocks that most influenced soil pH, while the mining byproduct (MB) led to high K levels. Zinc availability was highest in the treatments with mining byproduct and Cu in soil fertilized with Chapada and mining byproduct.

Phosphorus extracted by ion exchange resins and mehlich-1 from oxisols (latosols) treated with different phosphorus rates and sources for varied soil-source contact periods

Despite the large number of studies addressing the quantification of phosphorus (P) availability by different extraction methods, many questions remain unanswered. The aim of this paper was to compare the effectiveness of the extractors Mehlich-1, Anionic Resin (AR) and Mixed Resin (MR), to determine the availability of P under different experimental conditions. The laboratory study was arranged in randomized blocks in a [(3 x 3 x 2) + 3] x 4 factorial design, with four replications, testing the response of three soils with different texture: a very clayey Red Latosol (LV), a sandy clay loam Red Yellow Latosol (LVA), and a sandy loam Yellow Latosol (LA), to three sources (triple superphosphate, reactive phosphate rock from Gafsa-Tunisia; and natural phosphate from Araxá-Minas Gerais) at two P rates (75 and 150 mg dm-3), plus three control treatments (each soil without P application) after four contact periods (15, 30, 60, and 120 days) of the P sources with soil. The soil acidity of LV and LVA was adjusted by raising base saturation to 60 % with the application of CaCO3 and MgCO3 at a 4:1 molar ratio (LA required no correction). These samples were maintained at field moisture capacity for 30 days. After the contact periods, the samples were collected to quantify the available P concentrations by the three extractants. In general, all three indicated that the available P-content in soils was reduced after longer contact periods with the P sources. Of the three sources, this reduction was most pronounced for triple superphosphate, intermediate for reactive phosphate, while Araxá phosphate was least sensitive to the effect of time. It was observed that AR extracted lower P levels from all three soils when the sources were phosphate rocks, while MR extracted values close to Mehlich-1 in LV (clay) and LVA (medium texture) for reactive phosphate. For Araxá phosphate, much higher P values were determined by Mehlich-1 than by the resins, because of the acidity of the extractor. For triple superphosphate, both resins extracted higher P levels than Mehlich-1, due to the consumption of this extractor, particularly when used for LV and LVA.

SEQUENTIAL EXTRACTION OF PHOSPHORUS BY MEHLICH-1 AND ION EXCHANGE RESIN FROM B HORIZONS OF FERRIC AND PERFERRIC LATOSOLS (OXISOLS)

In general, Latosols have low levels of available P, however, the influence of the parent material seems to be decisive in defining the pool and predominant form of P in these soils. This study evaluated P availability by extraction with Mehlich-1 (M-1) and Ion Exchange Resin (IER), from samples of B horizons of Ferric and Perferric Latosols developed from different parent materials. To this end, in addition to the physical and chemical characterization of soils, 10 sequential extractions were performed with M-1 and IER from samples of B horizons (depth between 0.8 and 1.0 m). Total contents of Ca, P, Fe, Al, and Ti were determined after digestion with nitric, hydrofluoric and perchloric acids. The effects of sequential P extractions on Fe oxides were also evaluated from the analyses of dithionite-citrate-bicarbonate and ammonium acid oxalate. The high similarity between contents of P accumulated after sequential extractions with M-1 and IER in soils developed on tuffite indicated a predominance of P-Ca. Higher contents of P after a single IER extraction show greater efficiency in P removal from highly weathered soils, as from the Latosols studied here. The P contents also show the high sensitivity of extractant M-1 in highly buffered soils. Furthermore, a single extraction with extractant M-1 or IER is not sufficient to estimate the amount of labile P in these soils.

Physiological responses of two tropical weeds to shade: II. Leaf gas exchange and nitrogen content

Ipomoea asarifolia (Desr.) Roem. & Schultz (Convolvulaceae) and Stachytarpheta cayennensis (Rich) Vahl. (Verbenaceae), two weeds found in pastures and crop areas in the Brazilian Amazonia, Brazil, were grown in controlled environment cabinets under high (800-1000 µmol m-² s-¹) and low (200-350 µmol m-² s-¹) light regimes during a 40-day period. The objective was to determine the effect of shade on photosynthetic features and leaf nitrogen content of I. asarifolia and S. cayennensis. High-irradiance grown I. asarifolia leaves had significantly higher dark respiration and light saturated rates of photosynthesis than low-irradiance leaves. No significant differences for these traits, between treatments, were observed in S. cayennensis. Low-irradiance leaves of both species displayed higher CO2 assimilation rates under low irradiance. High-irradiance grown leaves of both species had less nitrogen per unit of weight. Low-irradiance S. cayennensis had more nitrogen per unit of leaf area than high-irradiance plants; however, I. asarifolia showed no consistent pattern for this variable through time. For S. cayennensis, leaf nitrogen content and CO2 assimilation were inversely correlated to the amount of biomass allocated to developing reproductive structures. These results are discussed in relation to their ecological and weed management implications.

Remaining phosphorus and sodium fluoride pH in soils with different clay contents and clay mineralogies

The remaining phosphorus (Prem) has been used for estimating the phosphorus buffer capacity (PBC) of soils of some Brazilian regions. Furthermore, the remaining phosphorus can also be used for estimating P, S and Zn soil critical levels determined with PBC-sensible extractants and for defining P and S levels to be used not only in P and S adsorption studies but also for the establishment of P and S response curves. The objective of this work was to evaluate the effects of soil clay content and clay mineralogy on Prem and its relationship with pH values measured in saturated NaF solution (pH NaF). Ammonium-oxalate-extractable aluminum exerts the major impacts on both Prem and pH NaF, which, in turn, are less dependent on soil clay content. Although Prem and pH NaF have consistent correlation, the former has a soil-PBC discriminatory capacity much greater than pH NaF.

Performance of Holstein heifers fed sugarcane silages treated with urea, sodium benzoate or Lactobacillus buchneri

The objective of this work was to evaluate the performance of heifers fed sugarcane silages produced with and without additives. Thirty-two Holstein heifers were randomly assigned, in a block design, to evaluate rations (46% silage; 54% concentrate; 12% crude protein) containing silages treated with (fresh basis) urea (0.5%), sodium benzoate (0.1%) or Lactobacillus buchneri (3.64x10(5) cfu g-1 ). Inoculation with L. buchneri improved daily gain (1.24 vs. 0.94 kg day-1 ), and the addition of benzoate resulted in better feed conversion (7.6 vs. 9.4 kg of dry matter per kg of live weight), in relation to the untreated silage (control). Treatments did not affect dry matter intake (mean of 2.19% of live weight). Rations containing silages treated with benzoate or L. buchneri showed lower cost per kg of weight gain. Treatment with urea did not improve animal performance, but the cost per kg of weight gain was lower than that of the control ration.

Oxygen consumption of Litopenaeus vannamei juveniles in heterotrophic medium with zero water exchange

This work aimed at determining the dissolved oxygen consumption rate of Litopenaeus vannamei juveniles maintained in a microbial biofloc raceway system at high density with no aeration. Three 4 L bottles were filled for each treatment, sealed hermetically, and placed in an enclosed greenhouse raceway system. Four shrimp (13.2±1.42 g) were assigned to two sets of the bottles, which underwent the following treatments: light conditions with no shrimp; dark conditions with no shrimp; light conditions with shrimp; and dark conditions with shrimp. Dissolved oxygen content was measured every 10 min for 30 min. A quadratic behavior was observed in dissolved oxygen concentration over time. Significant differences for oxigen consumption were observed only at 10 and 20 min between shrimp maintained in the dark and those under light conditions. At 10 min, a higher value was observed in shrimp maintained under light, and at 20 min, in the dark. Significant differences between 10 and 20 min and between 10 and 30 min were observed when oxygen consumption was analyzed over time in the presence of light. Under dark conditions there were significant differences only between 20 and 30 min. Lethal oxygen concentration (0.65 mg L-1) would be reached in less than one hour either under light or dark conditions with no aeration.

Gas exchange rates, plant height, yield components, and productivity of upland rice as affected by plant regulators

The objective of this work was to evaluate gas exchange rates, plant height, yield components, and productivity of upland rice, as affected by type and application time of plant growth regulators. A randomized block design, in a 4x2 factorial arrangement, with four replicates was used. Treatments consisted of three growth regulators (mepiquat chloride, trinexapac-ethyl, and paclobutrazol), besides a control treatment applied at two different phenological stages: early tillering or panicle primordial differentiation. The experiment was performed under sprinkler-irrigated field conditions. Net CO2 assimilation, stomatal conductance, plant transpiration, and water-use efficiency were measured four times in Primavera upland rice cultivar, between booting and milky grain phenophases. Gas exchange rates were neither influenced by growth regulators nor by application time. There was, however, interaction between these factors on the other variables. Application of trinexapac-ethyl at both tillering and differentiation stages reduced plant height and negatively affected yield components and rice productivity. However, paclobutrazol and mepiquat chloride applied at tillering, reduced plant height without affecting rice yield. Mepiquat chloride acted as a growth stimulator when applied at the differentiation stage, and significantly increased plant height, panicle number, and grain yield of upland rice.