Immunological response to re-infections with clones of the Colombian strain of Trypanosoma cruzi with different degrees of virulence: influence on pathological features during chronic infection in mice

Re-infections with Trypanosoma cruzi are an aggravating factor for Chagas disease morbidity. The Colombian strain of T. cruzirepresents multiclonal populations formed by clonally propagating organisms with different tropisms and degrees of virulence. In the present study, the influence of successive inoculations with clones of the Colombian strain, exhibiting different degrees of virulence, on chronic myocarditis and the humoral and cellular immune responses (Col-C1 high virulence, Col-C8 medium virulence and Col-C5 low virulence) were demonstrated. Mice from three groups with a single infection were evaluated during the acute (14th-30th day) and chronic phases for 175 days. An immunofluorescence assay, ELISA and delayed type hypersensitivity (DTH) cutaneous test were also performed. Mice with a triple infection were studied on the 115th-175th days following first inoculation. The levels of IgM and IgG2a were higher in the animals with a triple infection. DTH showed a higher intensity in the inflammatory infiltrate based on the morphometric analysis during a 48 h period of the triple infection and at 24 h with a single infection. The histopathology of the heart demonstrated significant exacerbation of cardiac inflammatory lesions confirmed by the morphometric test. The humoral responses indicate a reaction to the triple infection, even with clones of the same strain.

Sistemática, filogenia e distribuição geográfica das espécies sul-americanas de Centris (Paracentris) Cameron, 1903 e de Centris (Penthemisia) Moure, 1950, incluindo uma análise filogenética do "grupo Centris" sensu Ayala, 1998 (Hymenoptera, Apoidea, Centridini)

Systematics, phylogeny and geographical distribution of the South American species of Centris (Paracentris) Cameron, 1903, and Centris (Penthemisia) Moure, 1950, including a phylogenetic analysis of the "Centris group" sensu Ayala, 1998 (Hymenoptera, Apoidea, Centridini). A cladistic analysis with the objective of testing the hypothesis of monophily of Centris (Paracentris) Cameron, 1903, and of studying its phylogenetic relationships with the other subgenera that belong to the Centris group, sensu Ayala, 1998, and the relationships among the species that occur in South America, is presented. Centris (Paracentris) is a group of New World bees of amphitropical distribution, especially diversified in the Andes and in the xeric areas of South and North America. Thirty-one species were included in the analysis, four considered as outgroup, and 49 characters, all from external morphology and genitalia of adult specimens. Parsimony analyses with equal weights for the characters and successive weighting were performed with the programs NONA and PAUP, and analyses of implied weighting with the program PeeWee. The strict consensus among the trees obtained in all the analyses indicates that C. (Paracentris), as previously recognized, is a paraphyletic group. In order to eliminate that condition, the subgenera C. (Acritocentris), C. (Exallocentris) and C. (Xerocentris), all described by SNELLING (1974) are synonymized under C. (Paracentris). The subgenus C. (Penthemisia) Moure, 1950, previously considered a synonym of C. (Paracentris), is reinstated, but in a more restricted sense than originally proposed and with the following species: Centris brethesi Schrottky, 1902; C. buchholzi Herbst, 1918; C. chilensis (Spinola, 1851), C. mixta mixta Friese, 1904, and C. mixta tamarugalis Toro & Chiappa, 1989. Centris mixta, previously recognized as the only South American species of the subgenus C. (Xerocentris), a group supposedly amphitropical, came out as the sister-species of C. buchholzi. The following South American species were recognized under Centris (Paracentris): Centris burgdorfi Friese, 1901; C. caelebs Friese, 1900; C. cordillerana Roig-Alsina, 2000; C. euphenax Cockerell, 1913; C. flavohirta Friese, 1900; C. garleppi (Schrottky, 1913); C. klugii Friese, 1900; C. lyngbyei Jensen-Haarup, 1908; C. mourei Roig-Alsina, 2000; C. neffi Moure, 2000; C. nigerrima (Spinola, 1851); C. toroi sp. nov.; C. tricolor Friese, 1900; C. unifasciata (Schrottky, 1913), and C. vogeli Roig-Alsina, 2000. The relationships among the subgenera of the "Centris group" were: (Xanthemisia (Penthemisia (Centris s. str. - Paracentris))). Centris xanthomelaena Moure & Castro 2001, an endemic species of the Caatinga and previously considered a C. (Paracentris), came out as the sister group of C. (Centris) s. str. A new species of C. (Paracentris) from Chile is described: Centris toroi sp. nov. Lectotypus designations and redescriptions are presented for Centris burgdorfi, C. caelebs, C. lyngbyei, C. tricolor, C. autrani Vachal, 1904 and C. smithii Friese, 1900. New synonyms proposed: C. buchholzi Herbst, 1918 = Centris wilmattae Cockerell, 1926 syn. nov.; C. caelebs Friese, 1900 = Paracentris fulvohirta Cameron, 1903. The female of C. vogeli Roig-Alsina, 2000 and the male of C. xanthomelaena are described.

Enhanced degradation of metalaxyl in Gley Humic and Dark Red Latosol

Enhanced degradation of the fungicide metalaxyl was investigated in two soils: a gley humic (GH) and a Dark Red Latosol (LE), collected at sites never exposed to the fungicide. The soil samples were treated with successive applications of metalaxyl as a commercial formulation and 14C-metalaxyl in laboratory. Metalaxyl biodegradation was analyzed during 63 days by means of radiometric techniques to verify biomineralization and degradation product formation from the applied 14C-metalaxyl. Although biomineralization (maximum of 14 and 8% in the GH and LE soils, respectively), and partial degradation (about 32 and 48%, respectively) were detected in both soils, enhanced degradation was verified only in the GH soil. Results proved that metalaxyl behaves differently in soils.

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.

Mobility of inorganic and organic phosphorus forms under different levels of phosphate and poultry litter fertilization in soils

The eutrophication of aquifers is strongly linked to the mobility of P in soils. Although P mobility was considered irrelevant in a more distant past, more recent studies have shown that P, both in organic (Po) and inorganic forms (Pi), can be lost by leaching and eluviation through the soil profile, particularly in less weathered and/or sandier soils with low P adsorption capacity. The purpose of this study was to determine losses of P forms by leaching and eluviation from soil columns. Each column consisted of five PVC rings (diameter 5 cm, height 10 cm), filled with two soil types: a clayey Red-Yellow Latosol and a sandy loam Red-Yellow Latosol, which were exposed to water percolation. The soils were previously treated with four P rates (as KH2PO4 ) to reach 0, 12.5, 25.0 and 50 % of the maximum P adsorption capacity (MPAC). The P source was homogenized with the whole soil volume and incubated for 60 days. After this period the soils were placed in the columns; the soil of the top ring was mixed with five poultry litter rates of 0, 20, 40, 80, and 160 t ha-1 (dry weight basis). Treatments consisted of a 4 x 5 x 2 factorial scheme corresponding to four MPAC levels, five poultry litter rates, two soils, with three replications, arranged in a completely randomized block design. Deionized water was percolated through the columns 10 times in 35 days to simulate about 1,200 mm rainfall. In the leachate of each column the inorganic P (reactive P, Pi) and organic P forms (unreactive P, Po) were determined. At the end of the experiment, the columns were disassembled and P was extracted with the extractants Mehlich-1 (HCl 0.05 mol L-1 and H2SO4 0.0125 mol L-1) and Olsen (NaHCO3 0.5 mol L-1; pH 8.5) from the soil of each ring. The Pi and Po fractions were measured by the Olsen extractant. It was found that under higher poultry litter rates the losses of unreactive P (Po) were 6.4 times higher than of reactive P (Pi). Both the previous P fertilization and increasing poultry litter rates caused a vertical movement of P down the soil columns, as verified by P concentrations extracted by Mehlich-1 and NaHCO3 (Olsen). The environmental critical level (ECL), i.e., the P soil concentration above which P leaching increases exponentially, was 100 and 150 mg dm-3 by Mehlich-1 and 40 and 60 mg dm-3 by Olsen, for the sandy loam and clay soils, respectively. In highly weathered soils, where residual P is accumulated by successive crops, P leaching through the profile can be significant, particularly when poultry litter is applied as fertilizer.

Copper, zinc and manganese in soils of two watersheds in Santa Catarina with intensive use of pig slurry

Systematic pig slurry application to crop soils may lead to the accumulation of heavy metals in regions with intensive pig raising. The aim of this study was to evaluate the accumulation of Cu, Zn and Mn in soils under systematic pig slurry application. For this purpose, soil samples were collected from two of the most representative watersheds of Santa Catarina where the predominant activity is pig raising. In each watershed, 12 properties were chosen to evaluate the different systems of pig husbandry (complete cycle (CC), farrowing (FaU) and finishing units (FiU)). Based on information of the producers, soil samples were collected in areas with and without systematic manure application. To determine the total Cu, Zn and Mn content in soils and manure, a methodology proposed by the Environmental Protection Agency of the United States (USEPA), method nº 3050B, was used. For the available heavy metal content, Cu and Zn was extracted with HCl 0.1 mol L-1 and Mn with KCl 1 mol L-1. Data were subjected to multivariate analysis, using the canonical discriminant analysis to identify the metals that best differentiate the soils studied within each swine housing system. Successive pig slurry applications cause an increase in Cu, Zn and Mn availability in the soil and this indicates the need for monitoring of the metal concentrations over time. The critical values of Cu in the soil can be reached and exceeded more rapidly than Zn. The results showed that the soil type may be one of the attribute underlying the determination of public policies in pig raising and waste management because soils such as Inceptisols were shown to be more prone to possible contamination since they may more rapidly reach total critical Cu levels.

Water retention in a peatland with organic matter in different decomposition stages

Peatlands are ecosystems formed by successive pedogenetic processes, resulting in progressive accumulation of plant remains in the soil column under conditions that inhibit the activity of most microbial decomposers. In Diamantina, state of Minas Gerais, Brazil, a peatland is located at 1366 m asl, in a region with a quartz-rich lithology and characteristic wet grassland vegetation. For this study, the peat area was divided in 12 transects, from which a total of 90 soil samples were collected at a distance of 20 m from each other. The properties rubbed fiber content (RF), bulk density (Bd), mineral material (MM), organic matter (OM), moisture (Moi) and maximum water holding capacity (MWHC) were analyzed in all samples. From three selected profiles of this whole area, samples were collected every 27 cm from the soil surface down to a depth of 216 cm. In these samples, moisture was additionally determined at a pressure of 10 kPa (Moi10) or 1500 kPa (Moi1500), using Richards' extractor and soil organic matter was fractionated by standard procedures. The OM decomposition stage of this peat was found to increase with soil depth. Moi and MWHC were highest in layers with less advanced stages of OM decomposition. The humin levels were highest in layers in earlier stages of OM decomposition and with higher levels of water retention at MWHC and Moi10. Humic acid contents were higher in layers at an intermediate stage of decomposition of organic matter and with lowest levels of water retention at MWHC, Moi10 and Moi1500.

Electrical conductivity of soil irrigated with swine wastewater estimated by time-domain reflectometry¹

Wastewater application to soil is an alternative for fertilization and water reuse. However, particular care must be taken with this practice, since successive wastewater applications can cause soil salinization. Time-domain reflectometry (TDR) allows for the simultaneous and continuous monitoring of both soil water content and apparent electrical conductivity and thus for the indirect measurement of the electrical conductivity of the soil solution. This study aimed to evaluate the suitability of TDR for the indirect determination of the electrical conductivity (ECse) of the saturated soil extract by using an empirical equation for the apparatus TDR Trase 6050X1. Disturbed soil samples saturated with swine wastewater were used, at soil proportions of 0, 0.45, 0.90, 1.80, 2.70, and 3.60 m³ m-3. The probes were equipped with three handmade 0.20 cm long rods. The fit of the empirical model that associated the TDR measured values of electrical conductivity (EC TDR) to ECse was excellent, indicating this approach as suitable for the determination of electrical conductivity of the soil solution.

Relations between soil surface roughness, tortuosity, tillage treatments, rainfall intensity and soil and water losses from a red yellow latosol

The soil surface roughness increases water retention and infiltration, reduces the runoff volume and speed and influences soil losses by water erosion. Similarly to other parameters, soil roughness is affected by the tillage system and rainfall volume. Based on these assumptions, the main purpose of this study was to evaluate the effect of tillage treatments on soil surface roughness (RR) and tortuosity (T) and to investigate the relationship with soil and water losses in a series of simulated rainfall events. The field study was carried out at the experimental station of EMBRAPA Southeastern Cattle Research Center in São Carlos (Fazenda Canchim), in São Paulo State, Brazil. Experimental plots of 33 m² were treated with two tillage practices in three replications, consisting of: untilled (no-tillage) soil (NTS) and conventionally tilled (plowing plus double disking) soil (CTS). Three successive simulated rain tests were applied in 24 h intervals. The three tests consisted of a first rain of 30 mm/h, a second of 30 mm/h and a third rain of 70 mm/h. Immediately after tilling and each rain simulation test, the surface roughness was measured, using a laser profile meter. The tillage treatments induced significant changes in soil surface roughness and tortuosity, demonstrating the importance of the tillage system for the physical surface conditions, favoring water retention and infiltration in the soil. The increase in surface roughness by the tillage treatments was considerably greater than its reduction by rain action. The surface roughness and tortuosity had more influence on the soil volume lost by surface runoff than in the conventional treatment. Possibly, other variables influenced soil and water losses from the no-tillage treatments, e.g., soil type, declivity, slope length, among others not analyzed in this study.

Phosphorus accumulation and pollution potential in a hapludult fertilized with pig manure

Successive applications of pig litter to the soil surface can increase the phosphorus (P) content and alter its adsorption, promoting P transfer to surface or subsurface waters. The purpose of this study was to evaluate P accumulation and the pollution potential of a soil after application of pig litter. In March 2010, eight years after the installation of an experiment in Braço do Norte, Santa Catarina, SC, Brazil, on a Typic Hapludult, soil was sampled (layers 0-2.5, 2.5-5, 5-10, 10-15, 15-20 and 20-30 cm) after the following fertilization treatments: no pig litter fertilization, pig slurry application and pig manure application. In this period, 694 and 1,890 kg P2O5 ha-1 were applied in the treatments with pig slurry and pig manure, respectively. The P content was determined, based on Mehlich-1, anion exchange resin (AER), 0.01 mol L-1 CaCl2 and total P in the samples. The adsorption isotherm parameters were also determined by the Langmuir and Koski-Vähälä & Hartikainem models in the layers 0-2.5 and 20-30 cm. The application of 1,890 kg P2O5 ha-1 in the form of pig manure led to P accumulation, as evidenced by Mehlich-1, down to a depth of 15 cm, by AER and 0.01 mol L-1 CaCl2 down to 20 cm and by total P to 30 cm. After application of 1,890 kg P2O5 ha-1 in the form of pig manure, the values of maximum P adsorption capacity were lowest in the deepest layer (20-30 cm), indicating the occupation of part of the adsorption sites of the particles. The application of swine manure to the soil over eight years increased the P quantity in the soil solution of the surface layer, indicating environmental contamination risk for surface and subsurface waters.

Changes in soil acidity and organic carbon in a sandy typic hapludalf after medium-term pig-slurry and deep-litter application

Successive applications of liquid swine waste to the soil can increase the contents of total organic carbon and nutrients and change acidity-related soil chemical properties. However, little information is available on the effects of swine waste application in solid form, as of swine deep-litter. The objective of this study was to evaluate alterations of organic carbon and acidity-related properties of a soil after eight years of pig slurry and deep-litter application. In the eighth year of a field experiment established in Braço do Norte, Santa Catarina (SC) on a sandy Typic Hapludalf samples were taken (layers 0-2.5; 2.5-5; 5-10; 10-15; 15-20 and 20-30 cm) from unfertilized plots and plots with pig slurry or deep-litter applications, providing the simple or double rate of N requirement of Zea mays and Avena strigosa in rotation. Soil total organic carbon, water pH, exchangeable Al, Ca and Mg, and cation exchange capacity (CECeffective and CECpH7.0), H+Al, base saturation, and aluminum saturation were measured. The application of pig slurry and deep-litter for eight years increased total organic carbon and CEC in all soil layers. The pig slurry and deep-litter applications reduced active acidity and aluminum saturation and increased base saturation down to a depth of 30 cm. Eight years of pig slurry application did not affect soil acidity.

Lead and zinc in the structure of organic and mineral soil components

In addition to the more reactive forms, metals can occur in the structure of minerals, and the sum of all these forms defines their total contents in different soil fractions. The isomorphic substitution of heavy metals for example alters the dimensions of the unit cell and mineral size. This study proposed a method of chemical fractionation of heavy metals, using more powerful extraction methods, to remove the organic and different mineral phases completely. Soil samples were taken from eight soil profiles (0-10, 10-20 and 20-40 cm) in a Pb mining and metallurgy area in Adrianópolis, Paraná, Brazil. The Pb and Zn concentrations were determined in the following fractions (complete phase removal in each sequential extraction): exchangeable; carbonates; organic matter; amorphous and crystalline Fe oxides; Al oxide, amorphous aluminosilicates and kaolinite; and residual fractions. The complete removal of organic matter and mineral phases in sequential extractions resulted in low participation of residual forms of Pb and Zn in the total concentrations of these metals in the soils: there was lower association of metals with primary and 2:1 minerals and refractory oxides. The powerful methods used here allow an identification of the complete metal-mineral associations, such as the occurrence of Pb and Zn in the structure of the minerals. The higher incidence of Zn than Pb in the structure of Fe oxides, due to isomorphic substitution, was attributed to a smaller difference between the ionic radius of Zn2+ and Fe3+.

Effects of nickel and nitrogen soil fertilization on lettuce growth and urease activity

Nickel is a micronutrient involved in nitrogen metabolism and a constituent of the urease molecule. Plant growth and urease activity were evaluated in lettuce (Lactuca sativa L.) grown in soil-filled pots in a 2 x 8 factorial design with two nitrogen (N) sources and eight Ni rates, with five replications. Nitrogen was applied at 200 mg dm-3 (half the dose incorporated into the soil at seedling transplanting and half top-dressed later) using the sources NH4NO3 (AN) and CO(NH2)2 (Ur). The Ni treatments (0, 2, 4, 8, 12, 16, 24 and 32 mg dm-3) were applied as NiCl2. The shoot dry-matter yield, leaf urease activity, Ni levels in the lettuce leaves and Ni levels extracted from soil with Mehlich-3 (M-3) and DTPA were determined. In the plants supplied with AN, the shoot dry-matter yield was higher than in those supplied with Ur. There was no difference in shoot dry matter in response to soil-applied Ni. The leaf urease activity increased with Ni application, regardless of the N source. The extractions with M-3 and DTPA were efficient to evaluate Ni availability for lettuce in the Red-Yellow Latosol.

Forms and accumulation of copper and zinc in a sandy typic hapludalf soil after long-term application of pig slurry and deep litter

Successive applications of pig slurry and pig deep litter may lead to an accumulation of copper (Cu) and zinc (Zn) fractions in the soil profile. The objective of this study was to evaluate the Cu and Zn forms and accumulation in a Sandy Typic Hapludalf soil after long-term application of pig slurry and deep litter. In March 2010, eight years after initiating an experiment in Braço do Norte, Santa Catarina (SC), Brazil, on a Sandy Typic Hapludalf soil, soil samples were collected from the 0-2.5, 2.5-5.0, 5-10 and 10-15 cm layers in treatments consisting of no manure application (control) and with applications of pig slurry and deep litter at two levels: the single and double rate of N requirement for maize and black oat succession. The soil was dried, ground in an agate mortar and analyzed for Cu and Zn contents by 0.01 mol L-1 EDTA and chemically fractionated to determine Cu and Zn. The applications of Pig deep litter and slurry at doses equivalent to 90 kg ha-1 N increased the contents of available Cu and Zn in the surface soil layer, if the double of this dose was applied in pig deep litter or double this dose in pig slurry, Cu and Zn migrated to a depth of 15 cm. Copper is accumulated mainly in the organic and residual fractions, and zinc preferentially in the fraction linked to clay minerals, especially in the surface soil layers.

Accumulation of phosphorus fractions and contamination potential in vineyard soils in the southern region of the state of Santa Catarina, Brazil

In vineyards, if phosphate is applied both before planting and at intervals during growth without consideration of technical criteria, the soil P fractions may be increased and their proportions altered. This study was carried out to evaluate the accumulation of P fractions and the parameters of the adsorption isotherm in a sandy Typic Hapludalf soil in vineyards with a history of successive and excessive phosphate fertilization. In December 2010, two vineyards were selected, one 4 and the other 15 years old, in Urussanga, State of Santa Catarina (Brazil). Three trenches were dug in each area and soil was collected from the 0-5, 5-10 and 10-20 cm depth ranges. The soil samples were dried in a forced-air oven, sieved and subjected to chemical analyses, P chemical fractionation and P adsorption isotherms. Excessive phosphate fertilization, before and during cultivation, particularly in the older vineyard and, consequently, with a longer history of phosphate fertilization, increased the inorganic P concentrations to the depth of 20 cm, especially in labile fractions extracted by anion exchange resin and NaHCO3 in the non-labile fraction, as well as in the non-labile fraction extracted by 1.0 mol L-1 HCl. The application of phosphate fertilizers and the long cultivation period increased the P levels in the organic labile fraction extracted by 0.5 mol L-1 NaHCO3, and especially in the moderately labile fraction extracted by 0.1 and 0.5 mol L-1 NaOH. Phosphate fertilization of older vineyards, i.e., cultivated for 15 years, increased the amounts of P desorbed in water, indicating a risk of contamination of surface waters and groundwater. The phosphate fertilization before planting, without considering the results of soil analysis, and during cultivation, disregarding the results of soil analysis, leaf analysis and expected yield, led to a reduction in the maximum P adsorption capacity in the 0-5 cm layer of vineyard 2, indicating saturation of part of the reactive particle adsorption sites.