Distinct chitinases are expressed during various growth phases of the human pathogen Paracoccidioides brasiliensis

The aim of this work was the partial purification and subsequent evaluation of chitinase expression during the various growth phases of Paracoccidioides brasiliensis. Initially, PbCTS1r was expressed as a recombinant protein and displayed enzymatic activity against 4-MU-[N-acetylglucosamine (GlcNAc)]3 and 4-MU-(GlcNAc)2. Two proteins, 45 kDa and 39 kDa in size, were partially purified from P. brasiliensis yeast crude extract using cation-exchange chromatography coupled with HPLC and were characterised as PbCTS1 and PbCTS2, respectively. Anti-PbCTS1r antibody recognised two proteins in the crude extracts of yeast and the transitional stage between mycelial and yeast phases. In crude extracts of mycelium, only the 45 kDa protein was detected. However, quantitative real-time polymerase chain reaction led to the detection of small quantities of Pbcts2 transcript in the mycelial phase. In the yeast cell wall extract, only the 39 kDa protein was detected. Moreover, both proteins were secreted by the yeast parasitic phase, suggesting that these proteins participate in the modulation of the fungal environment. Phylogenetic analysis of the predicted PbCTS1 and PbCTS2 proteins indicated that they code for distinct chitinases in P. brasiliensis. During evolution, P. brasiliensis could have acquired the paralogues Pbcts1 and Pbcts2 for growth and survival in diverse environments in both saprophytic and parasitic phases.

Cryptic species of Paracoccidioides brasiliensis: impact on paracoccidioidomycosis immunodiagnosis

We aimed to evaluate whether the occurrence of cryptic species of Paracoccidioides brasiliensis, S1, PS2, PS3 and Paracoccidioides lutzii, has implications in the immunodiagnosis of paracoccidioidomycosis (PCM). Small quantities of the antigen gp43 were found in culture filtrates of P. lutzii strains and this molecule appeared to be more variable within P. lutzii because the synonymous-nonsynonymous mutation rate was lower, indicating an evolutionary process different from that of the remaining genotypes. The production of gp43 also varied between isolates belonging to the same species, indicating that speciation events are important, but not sufficient to fully explain the diversity in the production of this antigen. The culture filtrate antigen AgEpm83, which was obtained from a PS3 isolate, showed large quantities of gp43 and reactivity by immunodiffusion assays, similar to the standard antigen (AgB-339) from an S1 isolate. Furthermore, AgEpm83 was capable of serologically differentiating five serum samples from patients from the Botucatu and Jundiaí regions. These patients had confirmed PCM but, were non-reactive to the standard antigen, thus demonstrating an alternative for serological diagnosis in regions in which S1 and PS2 occur. We also emphasise that it is not advisable to use a single antigen preparation to diagnose PCM, a disease that is caused by highly diverse pathogens.

Adherence to statin treatment and associated factors in female users from the Unified Health System (SUS)

Objective: To identify the adherence rate of a statin treatment and possible related factors in female users from the Unified Health System. Method: Seventy-one women were evaluated (64.2 ± 11.0 years) regarding the socio-economic level, comorbidities, current medications, level of physical activity, self-report of muscular pain, adherence to the medical prescription, body composition and biochemical profile. The data were analyzed as frequencies, Chi-Squared test, and Mann Whitney test (p<0.05). Results: 15.5% of women did not adhere to the medical prescription for the statin treatment, whose had less comorbidities (p=0.01), consumed less quantities of medications (p=0.00), and tended to be younger (p=0.06). Those patients also presented higher values of lipid profile (CT: p=0.01; LDL-c: p=0.02). Musculoskeletal complains were not associated to the adherence rate to the medication. Conclusion: The associated factors to adherence of dyslipidemic women to statin medical prescription were age, quantity of comorbidities and quantity of current medication.


Arthropods associated with pig carrion in two vegetation profiles of Cerrado in the State of Minas Gerais, Brazil

Forensic Entomology research has been concentrated in only a few localities of the "Cerrado" vegetation, the Brazilian Savannah. The present study had, as its objective, an examination of the diversity of arthropod fauna associated with the carcasses of Sus scrofa (Linnaeus) in this biome. The study was conducted during the dry and humid periods in two Cerrado vegetation profiles of the State of Minas Gerais. The decaying process was slower and greater quantities of arthropods were collected during the dry period. Insects represented 99% of 161,116 arthropods collected. The majority of these were Diptera (80.2%) and Coleoptera (8.8%). The entomofauna belong to 85 families and at least 212 species. Diptera were represented by 31 families and at least 132 species. Sarcophagidae (Diptera) and Scarabaeidae (Coleoptera) were the richest groups. Oxysarcodexia (Sarcophagidae) presented the largest number of attracted species, however none of these species bred in the carcasses. The Coleoptera collected belong to at least 50 species of 21 families. Among these species, Dermestes maculatus and Necrobia rufipes were observed breeding in the carcasses. This study showed species with potential importance for estimating the postmortem interval (PMI), indicative of seasonal and environmental type located.

Crepuscular activity of culicids (Diptera, Culicidae) in the peridomicile and in the remaining riparian forest in Tibagi river, State of Paraná, Brazil

Crepuscular activity of culicids (Diptera, Culicidae) in the peridomicile and in the remaining riparian forest in Tibagi river, State of Paraná, Brazil. Human-attracted mosquitoes were collected for one hour, around sunset time (half hour before and half after), from April to December 2006, in two environments (riparian forest and near houses), in Tibagi river basin, Palmeira municipality, State of Paraná. Seven-hundred forty-nine mosquitoes, belonging to 13 species, were collected. Psorophora champerico Dyar & Knab, 1906 (42.86%) and Psorophora discrucians (Walker, 1856) (40.59%) were the most frequent species. No significant differences between quantities of Ps. champerico (t = -0.792; d.f. = 16; p = 0.43) and Ps. discrucians (t = 0.689; d.f. = 16; p = 0.49) obtained in riparian forest and near houses were observed, indicating similar conditions for crepuscular activity of these species in both environments. Psorophora champerico and Ps. discrucians responded (haematophagic activity) to environmental stimuli associated with the twilight hours differently in distinct habitats studied. The former species is registered for the first time in the Atlantic forest biome.

Occurrence of adults and biological aspects of Geniates borelli Camerano (Coleoptera, Scarabaeidae, Rutelinae) in Aquidauana, MS, Brazil

Occurrence of adults and biological aspects of Geniates borelli Camerano (Coleoptera, Scarabaeidae, Rutelinae) in Aquidauana, Mato Grosso do Sul, Brazil. Due to the importance of Geniates borelli Camerano as a pest in many crops, studies were developed at Universidade Estadual de Mato Grosso do Sul, Aquidauana campus, MS, Brazil. Adults were collected with a light trap from January 2006 to December 2007. 3,320 adults were collected, and the highest quantities were obtained in October 2006 and November 2007, with 1,548 and 802 adults recorded, respectively. Collected adults were kept in plastic containers with soil and Brachiaria decumbens seedlings for oviposition. 535 eggs measuring 2.30 × 1.60 mm were obtained. As the embryonic development progressed, eggs increased in size to 3.00 × 2.70 mm, and this change occurred between 6 and 10 days after oviposition. The embryonic period lasted 17.9 days. The first, second, and third instars lasted 37.6, 49.7, and 74 days, respectively. The prepupal stage lasted 65.9 days and the pupal stage lasted an average of 18.5 days. The biological cycle is completed in 315.8 days, which characterizes the species as univoltine. The average longevity of females was 35.4 days and 28.5 days for males.

Biological aspects of Leucothyreus alvarengai Frey and Leucothyreus aff. semipruinosus Ohaus (Coleoptera, Melolonthidae, Rutelinae) in crop succession at central Brazil

Biological aspects of Leucothyreus alvarengai Frey and Leucothyreus aff. semipruinosus Ohaus (Coleoptera, Melolonthidae, Rutelinae) in crop succession at central Brazil. Beetles of the family Melolonthidae make up a large group and some species are considered pests of planted crops. Little information is available on the basic biological aspects of the genus Leucothyreus, such as association with cultivated crops and their occurrence periods. Therefore studies were developed in soybean and corn crops in Tangará da Serra, Mato Grosso, Brazil, with the objective of studying the occurrence and biological aspects of Leucothyreus alvarengai Frey and Leucothyreus aff. semipruinosus Ohaus. For acquisition of immature specimens of both species, in April 2011 sampling was performed in corn fields, in July and October in the fallow area, and in soybeans fields planted in December; in 2012 sampling was performed in January and February in soybean fields and in March in corn fields. In 2011 the total number of larvae obtained in April, July, October and December were 100, 6, 30 and 27, and in January, February and March of 2012 these quantities were 32, 52 and 65 larvae, respectively. In all sampling events the larvae of L. alvarengai were collected in greater quantity. At the beginning of the reproductive period of L. alvarengai and L. aff. semipruinosus, it was observed that the adults began to fly and soon after started oviposition in the field in September. The appearance of larvae coincides with the time of soybean planting in the field, thus the larvae feed on roots of soybean plants at the beginning of their development and the cycle from egg to adult of the two species was completed in one year.

Nutrient relations during an eucalyptus cycle at different population densities

To synchronize nutrient availability with the requirements of eucalyptus during a cultivation cycle, the nutrient flow of this system must be well understood. Essential, for example, is information about nutrient dynamics in eucalyptus plantations throughout a cultivation cycle, as well as impacts on soil nutrient reserves caused by the accumulation and subsequent export of nutrients via biomass. It is also important to quantify the effect of some management practices, such as tree population density (PD) on these fluxes. Some nutrient relations in an experiment with Eucalyptus grandis, grown at different PDs in Santa Barbara, state of Minas Gerais, Brazil, were evaluated for one cultivation cycle. At forest ages of 0.25, 2.5, 4.5, and 6.75 years, evaluations were carried out in the stands at seven different PDs (between 500 and 5,000 trees ha-1) which consisted in chemical analyses of plant tissue sampled from components of the aboveground parts of the tree, from the forest floor and the litterfall. Nutrient contents and allocations of the different biomass components were estimated. In general, there were only small and statistically insignificant effects of PD on the nutrient concentration in trees. With increasing forest age, P, K, Ca and Mg concentrations were reduced in the aboveground components and the forest floor. The magnitud of biochemical nutrient cycling followed the sequence: P > K > N > Mg. At the end of the cycle, the quantities of N, P, Ca and Mg immobilized in the forest floor were higher than in the other components.

Phosphorus transformation in poultry litter and litter-treated Oxisol of Brazil assessed by 31P-NMR and wet chemical fractionation

Large quantities of poultry litter are being produced in Brazil, which contain appreciable amounts of phosphorus (P) that could be of environmental concern. To assess the immediate environmental threat, five poultry litters composed of diverse bedding material were incubated for 43 days under greenhouse conditions. The litters consisted of: coffee bean husk (CH); wood chips (WC); rice husk (RH); ground corn cobs (CC) and ground napier grass (NG) (Pennisetum purpureum Schum.), in which the change in forms of soluble P was evaluated using 31P NMR spectroscopy. On average, 80.2 and 19.8 % of the total P in the extract, respectively, accounted for the inorganic and organic forms before incubation and 48 % of the organic P was mineralized to inorganic P in 43 days of incubation. Wide variation in the organic P mineralization rate (from 82 % -WC to 4 % - NG) was observed among litters. Inorganic orthophosphate (99.9 %) and pyrophosphate (0.1 %) were the only inorganic P forms, whereas the organic P forms orthophosphate monoesters (76.3 %) and diester (23.7 %) were detected. Diester P compounds were mineralized almost completely in all litters, except in the CH litter, within the incubation period. Pyrophosphates contributed with less than 0.5% and remained unaltered during the incubation period. Wood-chip litter had a higher organic P (40 %) content and a higher diester: monoester ratio; it was therefore mineralized rapidly, within the first 15 days, achieving steady state by the 29th day. Distinct mineralization patterns were observed in the litter when incubated with a clayey Oxisol. The substantial decrease observed in the organic P fraction (Po) of the litter types followed the order: CH (45 %) > CC (25 %) > RH (13 %) ≈ NG (12 %) > WC (5 %), whereas the Pi fraction increased. Incubation of RH litter in soil slowed down the mineralization of organic P.

Nutrient extraction and exportation by common bean cultivars under different fertilization levels: I - macronutrients

The use of cultivars with a higher yield potential and the adoption of new technology have achieved high grain yields in common bean, which probably changed the demand for nutrients in this crop. However, there is almost no information about the periods of the cycle in which nutrients are most demanded at which quantities by the main cultivars. The objective of this study was to evaluate the macronutrient extraction and exportation by the common bean cultivars Pérola and IAC Alvorada, under different levels of NPK fertilization, on a dystroferric Red Nitosol, in Botucatu, São Paulo State, Brazil. The experiment was arranged in a randomized complete block (split plot) design with four replications. The plots consisted of six treatments based on a 2 x 3 factorial model, represented by two cultivars and three NPK levels (PD0 - 'Pérola' without fertilization, PD1 - 'Pérola' with 50 % of recommended fertilization, PD2 - 'Pérola' with 100 % of recommended fertilization, AD0 - 'IAC Alvorada' without fertilization, AD1 - 'IAC Alvorada' with 50 % of recommended fertilization, and AD2 - 'IAC Alvorada' with 100 % of recommended fertilization) and subplots sampled seven times during the cycle. At higher levels of NPK fertilization, the grain yield and macronutrient extraction and exportation of both cultivars were higher, but without statistical differences. Macronutrient absorption was higher in the treatments with 100 % of recommended NPK fertilization (average amounts per hectare: 140 kg N, 16.5 kg P, 120 kg K, 69 kg Ca, 17.9 kg Mg, and 16.3 kg S). Regardless of the treatment, the demand for N, P, K, Ca, and Mg was highest from 45 to 55 days after emergence (DAE), i.e., in the R7 stage (pod formation), while the highest S absorption rates were concentrated between 55 and 65 DAE. More than 70 % of P, between 58 and 69 % of N, 40 and 52 % of S, 40 and 48 % of K, and 35 and 45 % of Mg absorbed during the cycle was exported with grains, whereas less than 15 % of Ca was exported.

Silicate fertilization of tropical soils: silicon availability and recovery index of sugarcane

Sugarcane is considered a Si-accumulating plant, but in Brazil, where several soil types are used for cultivation, there is little information about silicon (Si) fertilization. The objectives of this study were to evaluate the silicon availability, uptake and recovery index of Si from the applied silicate on tropical soils with and without silicate fertilization, in three crops. The experiments in pots (100 L) were performed with specific Si rates (0, 185, 370 and 555 kg ha-1 Si), three soils (Quartzipsamment-Q, 6 % clay; Rhodic Hapludox-RH, 22 % clay; and Rhodic Acrudox-RA, 68 % clay), with four replications. The silicon source was Ca-Mg silicate. The same Ca and Mg quantities were applied to all pots, with lime and/or MgCl2, when necessary. Sugarcane was harvested in the plant cane and first- and second-ratoon crops. The silicon rates increased soil Si availability and Si uptake by sugarcane and had a strong residual effect. The contents of soluble Si were reduced by harvesting and increased with silicate application in the following decreasing order: Q>RH>RA. The silicate rates promoted an increase in soluble Si-acetic acid at harvest for all crops and in all soils, except RA. The amounts of Si-CaCl2 were not influenced by silicate in the ratoon crops. The plant Si uptake increased according to the Si rates and was highest in RA at all harvests. The recovery index of applied Si (RI) of sugarcane increased over time, and was highest in RA.

Quantification of aluminium in soil of the Solimões Formation, Acre State, Brazil

The variety of soils in the State of Acre is wide and their chemical profiles are still not fully understood. The nature of the material of origin of these soils is indicated by the high aluminium (Al) content, commonly associated with high calcium (Ca) and magnesium (Mg) contents. The study objective was to use different methods to quantify Al in soils from toposequences formed from material of a sedimentary nature originating from the Solimões Formation, in Acre, Brazil. Trenches were opened at three distinct points in the landscape: shoulder, backslope and footslope positions. Soil samples were collected for physical, chemical, mineralogical analyses. The Al content was quantified using different methods. High Al contents were found in most of these horizons, associated with high Ca and Mg levels, representing the predominant cations in the sum of exchangeable bases. The mineralogy indicates that the soils are still in a low weathering phase, with the presence of significant quantities of 2:1 minerals. Similar Al contents were determined by the methods of NaOH titration, xylenol orange spectrometry and inductively coupled plasma optical emission spectrometry. However, no consistent data were obtained by the pyrocatechol violet method. Extraction with KCl overestimated the exchangeable Al content due to its ability to extract the non-exchangeable Al present in the smectite interlayers. It was observed that high Al contents are related to the instability of the hydroxyl-Al smectite interlayers.

Soluble carbon in oxisol under the effect of organic residue rates

The application of organic residues to the soil can increase soluble organic carbon (SOC) and affect the pH and electrolytic conductivity (EC) of the soil. However, the magnitude of these changes depends on the type of residue and the applied dose. This study aimed to evaluate the effect of increasing C rates contained in organic residue on the pH, EC, water-extractable total carbon (WETC), water-extractable organic carbon (WEOC), and water-extractable inorganic carbon (WEIC) in soil treated with manure (chicken, swine, and quail), sawdust, coffee husk, and sewage sludge. The levels of total C (TC- KH2PO4), organic carbon (OC- KH2PO4), and inorganic C (IC- KH2PO4) extractable by a 0.1 mol L-1 KH2PO4 solution were also quantified in soil under the effect of increasing rates of chicken and quail manures. The following rates of organic residue C were applied to a dystrophic Red Latosol (Oxisol) sample: 0, 2,000, 5,000, 10,000, and 20,000 mg kg-1. The addition of organic residues to the soil increased pH, except in the case of sewage sludge, which acidified the soil. The acidity correction potential of chicken and quail manure was highest, dependent on the manure rate applied; regardless of the dose used, sawdust barely alters the soil pH. At all tested rates, the EC of the soil treated with swine manure, coffee husk, and sawdust remained below 2.0 dS m-1, which is a critical level for salinity-sensitive crops. However, the application of chicken or quail manure and sewage sludge at certain rates increased the EC to values above this threshold level. Highest levels of WETC, WEOC, and WEIC were obtained when chicken and quail manure and coffee husk were applied to the Oxisol. The quantities of SOC extracted by KH2PO4 were higher than the quantities extracted by water, demonstrating the ability of soil to adsorb C into its colloids.

Crop response to organic fertilization with supplementary mineral nitrogen

Animal manure is applied to the soil as a nutrient source, especially of nitrogen, to plants. However, manure application rates can be reduced with the use of N fertilizer in topdressing. The aim of this study was to evaluate crop responses to different application rates of animal manure sources, used alone and supplemented with mineral N topdressing, in a no-tillage system. The study was carried out from 2005 to 2008 on a Hapludalf soil. The treatments consisted of rates of 10, 20 and 30 m³ ha-1 of pig slurry (PS), and of 1 and 2 t ha-1 of turkey manure (TM), applied alone and supplemented with topdressed N fertilizer (TNF), as well as two controls, mineral fertilization (NPK) and one control without fertilizer application. Grain yield in common bean and maize, and dry matter yield and nutrient accumulation in common bean, maize and black oat crops were evaluated. Nitrogen application in topdressing in maize and common bean, especially when PS was used at rates of 20 and 30 m³ ha-1, and TM, at 2 t ha-1, proved effective in increasing the crop grain yields, showing the viability of the combined use of organic and industrialized mineral sources. Nitrogen accumulation in maize and common bean tissues was the indicator most strongly related to grain yield, in contrast with the apparent nutrient recovery, which was not related to the N, P and K quantities applied in the organic sources. No clear residual effect of N topdressing of maize and common bean was observed on the dry matter yield of black oat grown in succession to the main crops with PS and TM applications.

Effects of pig slurry application on soil physical and chemical properties and glyphosate mobility

Pig slurry applied to soil at different rates may affect soil properties and the mobility of chemical compounds within the soil. The purpose of this study was to evaluate the effects of rates of pig slurry application in agricultural areas on soil physical and chemical properties and on the mobility of glyphosate through the soil profile. The study was carried out in the 12th year of an experiment with pig slurry applied at rates of 0 (control), 50, 100 and 200 m³ ha-1 yr-1 on a Latossolo Vermelho distrófico (Hapludox) soil. In the control, the quantities of P and K removed by harvested grains were replaced in the next crop cycle. Soil physical properties (bulk density, porosity, texture, and saturated hydraulic conductivity) and chemical properties (organic matter, pH, extractable P, and exchangeable K) were measured. Soil solution samples were collected at depths of 20, 40 and 80 cm using suction lysimeters, and glyphosate concentrations were measured over a 60-day period after slurry application. Soil physical and chemical properties were little affected by the pig slurry applications, but soil pH was reduced and P levels increased in the surface layers. In turn, K levels were increased in sub-surface layers. Glyphosate concentrations tended to decrease over time but were not affected by pig slurry application. The concentrations of glyphosate found in different depths show that the pratice of this application in agricultural soils has the potential for contamination of groundwater, especially when the water table is the surface and heavy rains occur immediately after application.