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.

Antagonistic activity expressed by Shigella sonnei: identification of a putative new bacteriocin

Bacteriocins are antibacterial, proteinaceous substances that mediate microbial dynamics. Bacteriocin production is a highly disseminated property among all major lineages of bacteria, including Shigella. In this paper, we addressed the purification and characterisation of a bacteriocin produced by a Shigella sonnei strain (SS9) isolated from a child with acute diarrhoea. The substance was purified through ammonium-sulphate precipitation and sequential steps of chromatography. The intracellular fraction obtained at 75% ammonium sulphate maintained activity following exposure to pH values from 1-11 and storage at -80ºC for more than two years and was inactivated by high temperatures and proteases. The molecular mass of the purified bacteriocin was determined by mass spectrometry to be 18.56 kDa. The N-terminal sequence of the bacteriocin did not match any other antibacterial proteins described. A putative new bacteriocin produced by S. sonnei has been detected. This bacteriocin may represent a newly described protein or a previously described protein with a newly detected function. Considering that SS9 expresses antagonism against other diarrhoeagenic bacteria, the bacteriocin may contribute to S. sonnei virulence and is potentially applicable to either preventing or controlling diarrhoeal disease.

Nitrogen fractions in the microbial biomass in soils of southern Brazil

The reaction of nitrogen compounds with ninhydrin can be used as an indicator of cytoplasmic materials released from microbial cells killed by fumigation. Total-N, ninhydrin-reactive-N (NR-N), ammonium-N (A-N), and α-amino-N in the microbial biomass of soils from the State of Rio Grande do Sul, Brazil, were determined, in 1996, in 0.5 mol L-1 K2SO4 extracts of fumigated and non-fumigated soils. Total-N varied from 20.3 to 104.4 mg kg-1 and the ninhydrin-reactive-N corresponded, in average, to 27% of this. The ninhydrin-reactive-N was made up of 67% ammonium-N and 33% aminoacids with the amino group at the α-carbon position. It was concluded that colorimetric analysis of NR-N and A-N may be used as a direct measure of microbial N in soil. This simple and rapid procedure is adequate for routine analyses.

Effect of water and citrate solubility on agronomic effectiveness of acidulated phosphates in three consecutive corn crops

In the process of phosphate rock acidulation, several impure P compounds may be formed along with the desirable Ca and NH4 phosphates. Such compounds normally reduce the content of water-soluble P and thus the agronomic effectiveness of commercial fertilizers. In order to study this problem, a greenhouse experiment consisting of three consecutive corn crops was conducted in samples of a Red-Yellow Latosol (Typical Hapludox) in a completely randomized design (6 x 2 x 2), with four replicates. Six commercial fertilizers were added to 2 kg of soil at a rate of 70 mg kg-1 P, based on the content of soluble P in neutral ammonium citrate plus water (NAC + H2O) of the fertilizers. Fertilizer application occurred either in the original form or leached to remove the water-soluble fraction, either by mixing the fertilizer with the whole soil in the pots or with only 1 % of its volume. The corn plants were harvested 40 days after emergence to determine the shoot dry matter and accumulated P. For the first crop and localized application, the elimination of water-soluble P from the original fertilizers resulted in less bioavailable P for the plants. For the second and third crops, the effects of P source, leaching and application methods were not as evident as for the first, suggesting that the tested P sources may have similar efficiencies when considering successive cropping. The conclusion was drawn that the water-insoluble but NAC-soluble fractions of commercial P fertilizers are not necessarily inert because they can provide P in the long run.

Nitrogen application timing and soil inorganic nitrogen dynamics under no-till oat/maize sequential cropping

The timing of N application to maize is a key factor to be considered in no-till oat/maize sequential cropping. This study aimed to evaluate the influence of pre-planting, planting and sidedress N application on oat residue decomposition, on soil N immobilisation and remineralisation and on N uptake by maize plants in no-till oat/maize sequential cropping. Undisturbed soil cores of 10 and 20 cm diameter were collected from the 0-15 cm layer of a no-till Red Latossol, when the oat cover crop was in the milk-grain stage. Two greenhouse experiments were conducted simultaneously. Experiment A, established in the 10 cm diameter cores and without plant cultivation, was used to asses N dynamics in soil and oat residues. Experiment B, established in the 20 cm diameter cores and with maize cultivation, was used to assess plant growth and N uptake. An amount of 6.0 Mg ha-1 dry matter of oat residues was spread on the surface of the cores. A rate of 90 kg N ha-1 applied as ammonium sulphate in both experiments was split in pre-planting, planting and sidedress applications as follows: (a) 00-00-00 (control), (b) 90-00-00 (pre-planting application, 20 days before planting), (c) 00-90-00 (planting application), (d) 00-30-60 (split in a planting and a sidedress application 31 days after emergence), (e) 00-00-00* (control, without oat residue) and (f) 90-00-00* (pre-planting application, without oat residue). The N concentration and N content in oat residues were not affected during decomposition by N fertilisation. Most of the fertiliser NH4+-N was converted into NO3--N within 20 days after application. A significant decrease in NO3--N contents in the 0-4 cm layer was observed in all treatments between 40 and 60 days after the oat residue placement on the soil surface, suggesting the occurrence of N immobilisation in this period. Considering that most of the inorganic N was converted into NO3- and that no immobilisation of the pre planting fertiliser N occurred at the time of its application, it was possible to conclude that pre-planting applied N was prone to losses by leaching. On the other hand, with split N applications, maize plants showed N deficiency symptoms before sidedress application. Two indications for fertiliser-N management in no-till oat/maize sequential cropping could be suggested: (a) in case of split application, the sidedress should be earlier than 30 days after emergence, and (b) if integral application is preferred to save field operations, this should be done at planting.

The effect of nitrogen and sulphur fertilization on yield and quality of kohlrabi (Brassica oleracea, L.)

In a greenhouse pot experiment with kohlrabi, variety Luna, we explored the joint effect of N (0.6 g N per pot = 6 kg of soil) and S in the soil (25-35-45 mg kg-1 of S) on yields, on N, S and NO3- content in tubers and leaves, and on alterations in the amino acids concentration in the tubers. S fertilisation had no effect on tuber yields. The ranges of N content in tubers and leaves were narrow (between 1.42-1.48 % N and 1.21-1.35 % N, respectively) and the effect of S fertilisation was insignificant. S concentration in the tubers ranged between 0.59 and 0.64 % S. S fertilisation had a more pronounced effect on the S concentration in leaf tissues where it increased from 0.50 to 0.58 or to 0.76 % S under the applied dose. The NO3- content was higher in tubers than in leaves. Increasing the S level in the soil significantly reduced NO3- concentrations in the tubers by 42.2-53.6 % and in the leaves by 8.8-21.7 %. Increasing the S content in the soil reduced the concentration of cysteine + methionine by 16-28 %. The values of valine, tyrosine, aspartic acid and serine were constant. In the S0, S1, and S2 treatments the levels of threonine, isoleucine, leucine, arginine, the sum of essential amino acids and alanine decreased from 37 to 9 %. The histidine concentration increased with increasing S fertilisation. S fertilisation of kohlrabi can be recommended to stabilize the yield and reduce the undesirable NO3- contained in the parts used for consumption.

Fertilizer 15N balance in a coffee cropping system: a case study in Brazil

Knowledge about the fate of fertilizer nitrogen in agricultural systems is essential for the improvement of management practices in order to maximize nitrogen (N) recovery by the crop and reduce N losses from the system to a minimum. This study involves fertilizer management practices using the 15N isotope label applied in a single rate to determine the fertilizer-N balance in a particular soil-coffee-atmosphere system and to deepen the understanding of N plant dynamics. Five replicates consisting of plots of about 120 plants each were randomly defined within a 0.2 ha coffee plantation planted in 2001, in Piracicaba, SP, Brazil. Nine plants of each plot were separated in sub-plots for the 15N balance studies and treated with N rates of 280 and 350 kg ha-1 during 2003/2004 and 2004/2005, respectively, both of them as ammonium sulfate enriched to a 15N abundance of 2.072 atom %. Plant shoots were considered as separate parts: the orthotropic central branch, productive branches, leaves of productive branches, vegetative branches, leaves of vegetative branches and fruit. Litter, consisting of dead leaves accumulated below the plant canopy, was measured by the difference between leaves at harvest and at the beginning of the following flowering. Roots and soil were sampled down to a depth of 1.0 at intervals of 0.2 m. Samples from the isotopic sub-plots were used to evaluate total N and 15N, and plants outside sub-plots were used to evaluate dry matter. Volatilization losses of NH3 were estimated using special collectors. Leaching of fertilizer-N was estimated from deep drainage water fluxes and 15N concentrations of the soil solution at 1 m soil depth. At the end of the 2-year evaluation, the recovery of 15N applied as ammonium sulfate was 19.1 % in aerial plant parts, 9.4 % in the roots, 23.8 % in the litter, 26.3 % in the fruit and 12.6 % remaining in the 0_1.0 m soil profile. Annual leaching and volatilization losses were very small (2.0 % and 0.9 %, respectively). After two years, only 6.2 % N were missing in the balance (100 %) which can be attributed to other non-estimated compartments and experimental errors. Results show that an enrichment of only 2 % atom 15N allows the study of the partition of fertilizer-N in a perennial crop such as coffee during a period of two years.

Genesis of the sombric horizon in ultisols (red argisols) in southern Santa Catarina, Brazil

Dark subsurface horizons, with properties similar to the sombric horizon characterized by the USA Soil Taxonomy, are frequent in Southern Brazil. The genesis of this horizon is controversial and poorly understood. This study aimed to describe the occurrence of sombric-like horizons in Ultisols in the South of Santa Catarina State, at low altitudes, and suggest possible processes of humus transference, accumulation and persistence in these horizons. Physical, chemical and mineralogical properties of four Ultisols were evaluated; three were sampled in a toposequence, and another representative one in an isolated profile (RSP). The dark subsurface horizons coincide with the AB and BA transitional genetic horizons; they are acid, low in base saturation, and have a similar clay mineralogy in all horizons. Very high amounts of Fe and Al extracted by ammonium oxalate and sodium pyrophosphate solution as well as maximum Al extracted by CuCl2 solution were observed in these dark subsurface horizons, indicating a possible migration of these elements in the form of organometallic complexes. The contents of Al plus ½ Fe extracted from the RSP soil horizons with ammonium oxalate indicated spodic materials in the sombric-like horizon, although the soil morphology was not compatible with Spodosols. Maximum contents of fine clay were also found in the sombric-like horizon, suggesting Fe and Al migration as clay-humic substances. However, the hypothesis that sombric-like horizons in these soils are a relict feature of a grass paleovegetation, different from the current dense seasonal forest, should not be discarded but investigated in further studies.

15N uptake fom manure and fertilizer sources by three consecutive crops under controlled conditions

There are several regions of the world where soil N analysis and/or N budgets are not used to determine how much N to apply, resulting in higher than needed N inputs, especially when manure is used. One such region is the North Central "La Comarca Lagunera", one of the most important dairy production areas of Mexico. We conducted a unique controlled greenhouse study using 15N fertilizer and 15N isotopic-labeled manure that was labeled under local conditions to monitor N cycling and recovery under higher N inputs. The manure-N treatment was applied only once and was incorporated in the soil before planting the first forage crop at an equivalent rate of 30, 60 and 120 Mg ha-1 dry manure. The 15N treatments were equivalent to 120 and 240 kg ha-1 (NH4)2SO4-N for each crop. The total N fertilizer for each N fertilized treatment were 360, and 720 kg ha-1 N. We found very low N recoveries: about 9 % from the manure N inputs, lower than the 22 to 25 % from the fertilizer N inputs. The manure N recovered belowground in soil and roots ranged from 82 to 88 %. The low recoveries of N by the aboveground and low soil inorganic nitrate (NO3-N) and ammonium (NH4-N) content after the third harvested suggested that most of the 15N recovered belowground was in the soil organic form. The losses from manure N inputs ranged from 3 to 11 %, lower than the 34 to 39 % lost from fertilizer N sources. Our study shows that excessive applications of manure or fertilizer N that are traditionally used in this region will not increase the rate of N uptake by aboveground compartment but will increase the potential for N losses to the environment.

Spodosols pedogenesis under barreiras formation and sandbank environments in the south of Bahia

Morphologically differentiated Spodosols usually occur in the Coastal Plain of the South of Bahia and North of Espírito Santo. They are found in profiles known as "muçungas", i.e. sandy soils that accumulate water. In these areas, two kinds of Spodosols, different from those in the Restinga area, can be found: Spodosols with E albic horizon (white muçunungas) and without this horizon (black muçunungas). Eight soil profiles with spodic characteristics were collected and described in order to evaluate differences in the formation process of Barreiras and Restinga Spodosols in the South of Bahia. The soil profiles were also characterized chemically, physically and mineralogically. Additionally, texture and chemical analysis, Fe and Al extraction by sodium dithionite-citrate-bicarbonate (DBC), acid ammonium oxalate and sodium pyrophosphate, ammonium oxalate extract optic density (DOox), sulphuric acid attack, and X ray difractometry of the clay fraction were performed. In the Spodosols of the Barreiras area, fragipan was found the spodic layers. Cemented B spodic horizon were observed in the white muçunungas, and granular structure and dark color from the surface in the black muçunungas. There was no fragipan or hard spodic horizon in the Restinga Spodosol. This soil is acid, dystrophic and alic, with sandy texture and high clay percentages in the spodic horizons. The CEC, based on H + Al, is predominantly represented by the organic matter. The most representative components of the mineral phase of the clay fraction are kaolinite and possibly vermiculite traces with interlayered hydroxy. Chemical, physical, morphological and mineralogical differences were observed between the Barreiras and Restinga environments. The black and white muçunungas differ in morphologic and chemical properties only.

Nitrous oxide and methane fluxes in south Brazilian gleysol as affected by nitrogen fertilizers

Nitrogen fertilizers increase the nitrous oxide (N2O) emission and can reduce the methane (CH4) oxidation from agricultural soils. However, the magnitude of this effect is unknown in Southern Brazilian edaphoclimatic conditions, as well as the potential of different sources of mineral N fertilizers in such an effect. The aim of this study was to investigate the effects of different mineral N sources (urea, ammonium sulphate, calcium nitrate, ammonium nitrate, Uran, controlled- release N fertilizer, and urea with urease inhibitor) on N2O and CH4 fluxes from Gleysol in the South of Brazil (Porto Alegre, RS), in comparison to a control treatment without a N application. The experiment was arranged in a randomized block with three replications, and the N fertilizer was applied to corn at the V5 growth stage. Air samples were collected from a static chambers for 15 days after the N application and the N2O and CH4 concentration were determined by gas chromatography. The topmost emissions occurred three days after the N fertilizer application and ranged from 187.8 to 8587.4 µg m-2 h-1 N. The greatest emissions were observed for N-nitric based fertilizers, while N sources with a urease inhibitor and controlled release N presented the smallest values and the N-ammonium and amidic were intermediate. This peak of N2O emissions was related to soil NO3--N (R² = 0.56, p < 0.08) when the soil water-filled pore space was up to 70 % and it indicated that N2O was predominantly produced by a denitrification process in the soil. Soil CH4 fluxes ranged from -30.1 µg m-2 h-1 C (absorption) to +32.5 µg m-2 h-1 C (emission), and the accumulated emission in the period was related to the soil NH4+-N concentration (R² = 0.82, p < 0.001), probably due to enzymatic competition between nitrification and metanotrophy processes. Despite both of the gas fluxes being affected by N fertilizers, in the average of the treatments, the impact on CH4 emission (0.2 kg ha-1 equivalent CO2-C ) was a hundredfold minor than for N2O (132.8 kg ha-1 equivalent CO2-C). Accounting for the N2O and CH4 emissions plus energetic costs of N fertilizers of 1.3 kg CO2-C kg-1 N regarding the manufacture, transport and application, we estimated an environmental impact of N sources ranging from 220.4 to 664.5 kg ha-1 CO2 -C , which can only be partially offset by C sequestration in the soil, as no study in South Brazil reported an annual net soil C accumulation rate larger than 160 kg ha-1 C due to N fertilization. The N2O mitigation can be obtained by the replacement of N-nitric sources by ammonium and amidic fertilizers. Controlled release N fertilizers and urea with urease inhibitor are also potential alternatives to N2O emission mitigation to atmospheric and systematic studies are necessary to quantify their potential in Brazilian agroecosystems.

Ammonia volatilization in no-till system in the south-central region of the State of Paraná, Brazil

Ammonia (NH3) volatilization can reduce the efficiency of urea applied to the surface of no-till (NT) soils. Thus, the objectives of this study were to evaluate the magnitude of NH3 losses from surface-applied urea and to determine if this loss justifies the urea incorporation in soil or its substitution for other N sources under the subtropical climatic conditions of South-Central region of Paraná State, Brazil. The experiment, performed over four harvesting seasons in a clayey Hapludox followed a randomized block design with four replicates. A single dose of N (150 kg ha-1) to V5 growth stage of corn cultivated under NT system was applied and seven treatments were evaluated, including surface-applied urea, ammonium sulfate, ammonium nitrate, urea with urease inhibitor, controlled-release N source, a liquid N source, incorporated urea, and a control treatment with no N application. Ammonia volatilization was evaluated for 20 days after N application using a semi-open static system. The average cumulative NH3 loss due to the superficial application of urea was low (12.5 % of the applied N) compared to the losses observed in warmer regions of Southeastern Brazil (greater than 50 %). The greatest NH3 losses were observed in dry years (up to 25.4 % of the applied N), and losses decreased exponentially as the amount of rainfall after N application increased. Incorporated urea and alternative N sources, with the exception of controlled-release N source, decreased NH3 volatilization in comparison with surface-applied urea. Urea incorporation is advantageous for the reduction of NH3 volatilization; however, other aspects as its low operating efficiency should be considered before this practice is adopted. In the South-Central region of Paraná, the low NH3 losses from the surface-applied urea in NT system due to wet springs and mild temperatures do not justify its replacement for other N sources.

Recovery of degraded pasture in Rondônia: macronutrients and productivity of brachiaria brizantha

Pasture is the main form of land use in Amazonia. Over time the pasture grass loses vigor and yields decrease, indicating a certain degree of degeneration. The main causes of degradation are lack of pasture maintenance and subsequent weed infestation, the choice of regionally unsuitable forage species and excessive grazing. The main purpose of this study was to evaluate the impact of different recovery managements on soil chemical properties and grass yield of a degraded pasture in Rondônia. For this purpose, an experiment was installed in October 2001, consisting of five treatments: C = control; HA = harrowing + NPK + micronutrients; HE = Herbicide + NK + micronutrients; R = No-tillage rice + NPK + micronutrients; and S = No-tillage soybean + PK + micronutrients. The following N, P and K sources were used: ammonium sulfate for N, calcined phosphate for P and potassium chloride for K. The experiment was arranged in a randomized block design with four replications. The shoot dry matter yield of the grass was analyzed as of the 35th month of experimentation, in a dry and a rainy period. Phosphorus fertilization resulted in significant increases in Ca2+ and Mg2+ and increasing trend of P in the topsoil in the initial months of the experiment in treatments HA and S and increases in Ca2+ and P (trend) in the treatment R. The cumulative production of Brachiaria brizantha, from Sep/2004 to Mar/2005, was 30,025, 28,267 and 27,735 kg ha-1 shoot dry matter in the treatments HA, R and S, respectively. These values differed significantly from treatments C and HE, with 17,040 and 17,057 kg ha-1, respectively. It was concluded that phosphorus fertilization associated to pasture reform was effective to raise the dry matter yield of Brachiaria brizantha. Rice or soybean under no-tillage is recommended as a practice of pasture recovery, due to the residual effect of fertilization.

Nitrogen washing from C3 and C4 cover grasses residues by rain

Crop species with the C4 photosynthetic pathway are more efficient in assimilating N than C3 plants, which results in different N amounts prone to be washed from its straw by rain water. Such differences may affect N recycling in agricultural systems where these species are grown as cover crops. In this experiment, phytomass production and N leaching from the straw of grasses with different photosynthetic pathways were studied in response to N application. Pearl millet (Pennisetum glaucum) and congo grass (Brachiaria ruziziensis) with the C4 photosynthetic pathway, and black oat (Avena Strigosa) and triticale (X Triticosecale), with the C3 photosynthetic pathway, were grown for 47 days. After determining dry matter yields and N and C contents, a 30 mm rainfall was simulated over 8 t ha-1 of dry matter of each plant residue and the leached amounts of ammonium and nitrate were determined. C4 grasses responded to higher fertilizer rates, whereas N contents in plant tissue were lower. The amount of N leached from C4 grass residues was lower, probably because the C/N ratio is higher and N is more tightly bound to organic compounds. When planning a crop rotation system it is important to take into account the difference in N release of different plant residues which may affect N nutrition of the subsequent crop.

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.