Preliminary characterization of a Rhodnius prolixus hemolymph trypanolytic protein, this being a determinant of Trypanosoma rangeli KP1(+) and KP1(-) subpopulations' vectorial ability

Rhodnius prolixus is the main Trypanosoma rangeli vector in several Latin-American countries and is susceptible to infection with KP1(+) strains; however, it presents an invasion-resistant response to KP1(-) strains. The present work has identified a trypanolytic protein against T. rangeli KP1(-) in the R. prolixus hemolymph which was fractioned with ammonium sulfate (following dialysis). The results revealed a protein component which did not depend on divalent cations for its biological function whilst keeping its trypanolytic activity at temperatures ranging from -20ºC to 37ºC, at 7.0 to 10.5 pH. The protein was partially purified by gel filtration chromatography and ionic exchange chromatography. The major component presented a molecular weight of around 79 kDa and an isoelectric point between 4.9 and 6.3 and may be directly related to hemolymph trypanolytic activity against T. rangeli KP1(-) populations.

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.

Anti-flagellin antibody responses elicited in mice orally immunized with attenuated Salmonella enterica serovar Typhimurium vaccine strains

In the present study we investigated the flagellin-specific serum (IgG) and fecal (IgA) antibody responses elicited in BALB/c mice immunized with isogenic mutant derivatives of the attenuated Salmonella enterica serovar Typhimurium (S. Typhimurium) SL3261 strain expressing phase 1 (FliCi), phase 2 (FljB), or no endogenous flagellin. The data reported here indicate that mice orally immunized with recombinant S. Typhimurium strains do not mount significant systemic or secreted antibody responses to FliCi, FljB or heterologous B-cell epitopes genetically fused to FliCi. These findings are particularly relevant for those interested in the use of flagellins as molecular carriers of heterologous antigens vectored by attenuated S. Typhimurium strains.

Detection of Toxoplasma gondii oocysts in water: proposition of a strategy and evaluation in Champagne-Ardenne Region, France

Water is a vehicle for disseminating human and veterinary toxoplasmosis due to oocyst contamination. Several outbreaks of toxoplasmosis throughout the world have been related to contaminated drinking water. We have developed a method for the detection of Toxoplasma gondii oocysts in water and we propose a strategy for the detection of multiple waterborne parasites, including Cryptosporidium spp. and Giardia. Water samples were filtered to recover Toxoplasma oocysts and, after the detection of Cryptosporidium oocysts and Giardia cysts by immunofluorescence, as recommended by French norm procedure NF T 90-455, the samples were purified on a sucrose density gradient. Detection of Toxoplasma was based on PCR amplification and mouse inoculation to determine the presence and infectivity of recovered oocysts. After experimental seeding assays, we determined that the PCR assay was more sensitive than the bioassay. This strategy was then applied to 482 environmental water samples collected since 2001. We detected Toxoplasma DNA in 37 environmental samples (7.7%), including public drinking water; however, none of them were positive by bioassay. This strategy efficiently detects Toxoplasma oocysts in water and may be suitable as a public health sentinel method. Alternative methods can be used in conjunction with this one to determine the infectivity of parasites that were detected by molecular methods.

Absence of Fas-L aggravates renal injury in acute Trypanosoma cruzi infection

Trypanosoma cruzi infection induces diverse alterations in immunocompetent cells and organs, myocarditis and congestive heart failure. However, the physiological network of disturbances imposed by the infection has not been addressed thoroughly. Regarding myocarditis induced by the infection, we observed in our previous work that Fas-L-/- mice (gld/gld) have very mild inflammatory infiltration when compared to BALB/c mice. However, all mice from both lineages die in the early acute phase. Therefore, in this work we studied the physiological connection relating arterial pressure, renal function/damage and cardiac insufficiency as causes of death. Our results show that a broader set of dysfunctions that could be classified as a cardio/anaemic/renal syndrome is more likely responsible for cardiac failure and death in both lineages. However, gld/gld mice had very early glomerular deposition of IgM and a more intense renal inflammatory response with reduced renal filtration, which is probably responsible for the premature death in the absence of significant myocarditis in gld/gld.

The significance of the amoebocyte-producing organ in Biomphalaria glabrata

In molluscs, internal defence against microorganisms is performed by a single cell type, i.e., the haemocyte or amoebocyte. The origin of these cells in Biomphalaria glabrata was initially thought to be localised within the vasculo-connective tissue. More recently, origin from a single organ, termed the amoebocyte-producing organ (APO), has been postulated based on the occurrence of hyperplasia and mitoses during Schistosoma mansoni infection. The present investigation represents a histological, immuno-histochemical and ultra-structural study of the B. glabrata APO, whereby histological identification was facilitated by means of collecting epithelial basophilic cells. These cells were comprised of single-cell layers that cover a portion of the stroma, which contains many small, round cells and haemolymph sinuses, as well as a small area of the pericardial surface of the reno-pericardial region. On occasion, this epithelial component vaguely resembled the vertebrate juxtaglomerular apparatus, which reinforces its presumed relationship to the kidney. Both in normal and infected molluscs, mitoses were only occasionally found. The present quantitative studies failed to demonstrate the presence of APO cellular hyperplasia, either in normal or schistosome-infected B. glabrata. Conversely, several structural details from the APO region in B. glabrata were found to be consistent with the hypothesis that the APO is a filtration organ, i.e., it is more closely related to the kidney rather than the bone marrow, as has been suggested in the literature.

Growth and metabolic activity of the extramatrical mycelium of endomycorrhizal maize plants

The objective of this experiment was to quantify the extramatrical mycelium of the arbuscular mycorrhizal (AM) fungus Glomus etunicatum (Becker & Gerdemann) grown on maize (Zea mays L. var. Piranão) provided with various levels of phosphate fertilizer and harvested at 30, 60 and 90 days after planting (DAP). Total extramatrical mycelium (TEM) was extracted from soil using a modified membrane filtration method, followed by quantification using a grid intersection technique. Active extramatrical mycelium (AEM) proportion was determined using an enzymatic method which measured dehydrogenase activity by following iodonitrotetrazolium reduction. At low levels of added P, there was relatively less TEM than at high levels of added P, but the AEM proportion at low soil P availability was significantly greater than at high soil P.

Modelagem numérica para o processo da evaporação da água do solo

Foram realizados vários ensaios laboratoriais para avaliar o desempenho de um modelo numérico em simular o processo unidimensional da evaporação da água do solo. Este modelo foi desenvolvido a partir da linearização da equação de Richards no espaço e do uso da técnica iterativa de Newton-Raphson, para resolver essa equação não-linear no tempo, e leva em conta o fluxo de vapor dentro do perfil de solo e a taxa de evaporação na sua superfície. Os resultados mostraram que o modelo foi capaz de simular satisfatoriamente o processo, tanto em meios porosos homogêneos quanto estratificados

Nodulação e produção do caupi (Vigna unguiculata L. Walp) sob efeito de plantas de cobertura e inoculação

Um experimento de campo foi realizado no município de Crato - Ceará para estudar como a planta de cobertura afeta a nodulação e produtividade de caupi (Vigna unguiculata (L.) Walp), submetido a diferentes inoculações. Empregou-se o delineamento em blocos casualizados, no esquema em parcelas subsubdivididas, com quatro repetições. Os tratamentos consistiram de plantas de cobertura, formando a parcela principal; inoculação, como subparcela; e amostragens quinzenais, como subsubparcela. As plantas de cobertura foram: milheto - Pennisetum americanum; mucuna-preta - Stizolobium aterrimum ou vegetação espontânea, enquanto as fontes de N foram: sem inoculação; sem inoculação + 50 kg ha-1 de N na forma de uréia; inoculação com estirpes rizobianas (Bradyrhizobium sp.) recomendadas pela RELARE -BR 3301 + BR 3302; ou pela UFRPE - NFB 6156 + NFB 700. Foi utilizada a cultivar Patativa de caupi, recomendada para a região. A partir de sete dias após a emergência e até a maturidade final da cultura, foram realizadas amostras quinzenais, colhendo-se duas plantas, por subparcela para determinação do número de nódulos, matéria seca de nódulos e matéria seca da parte aérea. A partir dos 60 dias após emergência, foram realizadas colheitas semanais da área útil da subparcela, para determinação da produtividade de grãos. A mucuna aumentou a nodulação do caupi não-inoculado. A inoculação do caupi com estirpes recomendadas, ou o aumento da população nativa, permitiu produção de biomassa e grãos semelhante da adubação com 50 kg ha-1 de N.

Carbon dioxide efflux in a rhodic hapludox as affected by tillage systems in southern Brazil

Agricultural soils can act as a source or sink of atmospheric C, according to the soil management. This long-term experiment (22 years) was evaluated during 30 days in autumn, to quantify the effect of tillage systems (conventional tillage-CT and no-till-NT) on the soil CO2-C flux in a Rhodic Hapludox in Rio Grande do Sul State, Southern Brazil. A closed-dynamic system (Flux Chamber 6400-09, Licor) and a static system (alkali absorption) were used to measure soil CO2-C flux immediately after soybean harvest. Soil temperature and soil moisture were measured simultaneously with CO2-C flux, by Licor-6400 soil temperature probe and manual TDR, respectively. During the entire month, a CO2-C emission of less than 30 % of the C input through soybean crop residues was estimated. In the mean of a 30 day period, the CO2-C flux in NT soil was similar to CT, independent of the chamber type used for measurements. Differences in tillage systems with dynamic chamber were verified only in short term (daily evaluation), where NT had higher CO2-C flux than CT at the beginning of the evaluation period and lower flux at the end. The dynamic chamber was more efficient than the static chamber in capturing variations in CO2-C flux as a function of abiotic factors. In this chamber, the soil temperature and the water-filled pore space (WFPS), in the NT soil, explained 83 and 62 % of CO2-C flux, respectively. The Q10 factor, which evaluates CO2-C flux dependence on soil temperature, was estimated as 3.93, suggesting a high sensitivity of the biological activity to changes in soil temperature during fall season. The CO2-C flux measured in a closed dynamic chamber was correlated with the static alkali adsorption chamber only in the NT system, although the values were underestimated in comparison to the other, particularly in the case of high flux values. At low soil temperature and WFPS conditions, soil tillage caused a limited increase in soil CO2-C flux.

Programa de ajuste multiparamétrico de curvas de titulação potenciométricas de ácidos húmicos

Tendo em vista a dificuldade de determinação dos pontos de inflexão na curva de titulação potenciométrica de ácidos húmicos, por metodologias tradicionais, foi desenvolvido foi desenvolvido um programa na linguagem Delphi para ajuste multiparamétrico de dados de titulação potenciométrica. Para isso um processo iterativo para estimar as raízes de um polinômio, com base no método de Newton-Raphson, foi utilizado. Os dados das titulações potenciométricas de ácidos húmicos usados nas regressões foram obtidos em um sistema automatizado de titulação potenciométrica. O programa desenvolvido na linguagem Delphi permite maior versatilidade e facilidade de operação, com uma interação mais amigável com o usuário. As curvas de titulação potenciométricas ajustadas sobrepuseram-se quase que totalmente às curvas experimentais. Os valores de pKa e as percentagens de grupos tituláveis dos ácidos húmicos, parâmetros ajustáveis na regressão multiparamétrica, apresentaram valores comparáveis com dados da literatura.

Influence of liming on residual soil respiration and chemical properties in a tropical no-tillage system

Because of the climate changes occurring across the planet, especially global warming, the different forms of agricultural soil use have attracted researchers´ attention. Changes in soil management may influence soil respiration and, consequently, C sequestration. The objectives of this study were to evaluate the long-term influence of liming on soil respiration and correlate it with soil chemical properties after two years of liming in a no-tillage system. A randomized complete block design was used with six replications. The experimental treatments consisted of four lime rates and a control treatment without lime. Two years after liming, soil CO2 emission was measured and the soil sampled (layers 0-5, 5-10, 10-20, and 20-30 cm). The P, Ca2+ e Mg2+ soil contents and pH and base saturation were determined. CO2 emission from soil limed at the recommended rate was 24.1 % higher, and at twice the recommended rate, 47.4 % higher than from unlimed soil. Liming improved the chemical properties, and the linear increase in soil respiration rate correlated positively with the P, Ca2+ and Mg2+ soil contents, pH and base saturation, and negatively with H + Al and Al3+ contents. The correlation coefficient between soil respiration rate and chemical properties was highest in the 10-20 cm layer.

Magnetic susceptibility to identify landscape segments on a detailed scale in the region of Jaboticabal, São Paulo, Brazil

The agricultural potential is generally assessed and managed based on a one-dimensional vision of the soil profile, however, the increased appreciation of sustainable production has stimulated studies on faster and more accurate evaluation techniques and methods of the agricultural potential on detailed scales. The objective of this study was to investigate the possibility of using soil magnetic susceptibility for the identification of landscape segments on a detailed scale in the region of Jaboticabal, São Paulo State. The studied area has two slope curvatures: linear and concave, subdivided into three landscape segments: upper slope (US, concave), middle slope (MS, linear) and lower slope (LS, linear). In each of these segments, 20 points were randomly sampled from a database with 207 samples forming a regular grid installed in each landscape segment. The soil physical and chemical properties, CO2 emissions (FCO2) and magnetic susceptibility (MS) of the samples were evaluated represented by: magnetic susceptibility of air-dried fine earth (MS ADFE), magnetic susceptibility of the total sand fraction (MS TS) and magnetic susceptibility of the clay fraction (MS Cl) in the 0.00 - 0.15 m layer. The principal component analysis showed that MS is an important property that can be used to identify landscape segments, because the correlation of this property within the first principal component was high. The hierarchical cluster analysis method identified two groups based on the variables selected by principal component analysis; of the six selected variables, three were related to magnetic susceptibility. The landscape segments were differentiated similarly by the principal component analysis and by the cluster analysis using only the properties with higher discriminatory power. The cluster analysis of MS ADFE, MS TS and MS Cl allowed the formation of three groups that agree with the segment division established in the field. The grouping by cluster analysis indicated MS as a tool that could facilitate the identification of landscape segments and enable the mapping of more homogeneous areas at similar locations.

Nodulação e eficiência da fixação do N2 em feijão-caupi por efeito da taxa do inóculo

O feijão-caupi (Vigna unguiculata (L.) Walp.) pode adquirir nitrogênio em quantidades adequadas para suprir suas necessidades, por meio do processo de fixação biológica do nitrogênio (FBN), quando associado com rizóbios específicos e eficientes. O objetivo deste trabalho foi avaliar o efeito da aplicação de diferentes taxas de inóculo na nodulação e FBN na cultivar BRS Pujante de feijão-caupi. O experimento foi conduzido em casa de vegetação, utilizando-se vasos plásticos com 2 kg de solo, em delineamento experimental de blocos ao acaso com sete tratamentos e quatro repetições. Os tratamentos constaram de cinco concentrações de células da estirpe de Bradyrhizobium sp. BR 3267 (células/semente), um tratamento sem inoculação e com adição de fertilizante nitrogenado (controle + N) e um controle sem inoculação e sem adição de fertilizante nitrogenado (controle). Foram avaliados o número e a massa seca de nódulos, a massa seca da parte aérea e o acúmulo de N na parte aérea. À medida que houve aumento da concentração de células na semente, elevaram-se os parâmetros da nodulação e fixação do N2. A aplicação da maior taxa de inóculo, que foi de 6,65 x 10(7) células da estirpe BR 3267/semente, promoveu aumento da massa seca da parte aérea por planta correspondente a 27 % da massa seca da parte aérea do tratamento controle e semelhante ao tratamento controle + N. A inoculação a partir da aplicação de 8 x 10(5) células da estirpe BR 3267/semente forneceu maior quantidade de N para as plantas, em relação à dos tratamentos controle e controle + N. A cultivar BRS Pujante foi beneficiada pela FBN quando inoculada com a estirpe BR 3267, proporcionalmente à taxa de inóculo.

Uncertainties in the prediction of spatial variability of soil CO2 emissions and related properties

The soil CO2 emission has high spatial variability because it depends strongly on soil properties. The purpose of this study was to (i) characterize the spatial variability of soil respiration and related properties, (ii) evaluate the accuracy of results of the ordinary kriging method and sequential Gaussian simulation, and (iii) evaluate the uncertainty in predicting the spatial variability of soil CO2 emission and other properties using sequential Gaussian simulations. The study was conducted in a sugarcane area, using a regular sampling grid with 141 points, where soil CO2 emission, soil temperature, air-filled pore space, soil organic matter and soil bulk density were evaluated. All variables showed spatial dependence structure. The soil CO2 emission was positively correlated with organic matter (r = 0.25, p < 0.05) and air-filled pore space (r = 0.27, p < 0.01) and negatively with soil bulk density (r = -0.41, p < 0.01). However, when the estimated spatial values were considered, the air-filled pore space was the variable mainly responsible for the spatial characteristics of soil respiration, with a correlation of 0.26 (p < 0.01). For all variables, individual simulations represented the cumulative distribution functions and variograms better than ordinary kriging and E-type estimates. The greatest uncertainties in predicting soil CO2 emission were associated with areas with the highest estimated values, which produced estimates from 0.18 to 1.85 t CO2 ha-1, according to the different scenarios considered. The knowledge of the uncertainties generated by the different scenarios can be used in inventories of greenhouse gases, to provide conservative estimates of the potential emission of these gases.