Prokaryotic cells: structural organisation of the cytoskeleton and organelles

For many years, prokaryotic cells were distinguished from eukaryotic cells based on the simplicity of their cytoplasm, in which the presence of organelles and cytoskeletal structures had not been discovered. Based on current knowledge, this review describes the complex components of the prokaryotic cell cytoskeleton, including (i) tubulin homologues composed of FtsZ, BtuA, BtuB and several associated proteins, which play a fundamental role in cell division, (ii) actin-like homologues, such as MreB and Mb1, which are involved in controlling cell width and cell length, and (iii) intermediate filament homologues, including crescentin and CfpA, which localise on the concave side of a bacterium and along its inner curvature and associate with its membrane. Some prokaryotes exhibit specialised membrane-bound organelles in the cytoplasm, such as magnetosomes and acidocalcisomes, as well as protein complexes, such as carboxysomes. This review also examines recent data on the presence of nanotubes, which are structures that are well characterised in mammalian cells that allow direct contact and communication between cells.

Variability and resistance mutations in the hepatitis C virus NS3 protease in patients not treated with protease inhibitors

The goal of treatment of chronic hepatitis C is to achieve a sustained virological response, which is defined as exhibiting undetectable hepatitis C virus (HCV) RNA levels in serum following therapy for at least six months. However, the current treatment is only effective in 50% of patients infected with HCV genotype 1, the most prevalent genotype in Brazil. Inhibitors of the serine protease non-structural protein 3 (NS3) have therefore been developed to improve the responses of HCV-infected patients. However, the emergence of drug-resistant variants has been the major obstacle to therapeutic success. The goal of this study was to evaluate the presence of resistance mutations and genetic polymorphisms in the NS3 genomic region of HCV from 37 patients infected with HCV genotype 1 had not been treated with protease inhibitors. Plasma viral RNA was used to amplify and sequence the HCV NS3 gene. The results indicate that the catalytic triad is conserved. A large number of substitutions were observed in codons 153, 40 and 91; the resistant variants T54A, T54S, V55A, R155K and A156T were also detected. This study shows that resistance mutations and genetic polymorphisms are present in the NS3 region of HCV in patients who have not been treated with protease inhibitors, data that are important in determining the efficiency of this new class of drugs in Brazil.

The efficiency of concentration methods used to detect enteric viruses in anaerobically digested sludge

The presence of enteric viruses in biosolids can be underestimated due to the inefficient methods (mainly molecular methods) used to recover the viruses from these matrices. Therefore, the goal of this study was to evaluate the different methods used to recover adenoviruses (AdV), rotavirus species A (RVA), norovirus genogroup II (NoV GII) and the hepatitis A virus (HAV) from biosolid samples at a large urban wastewater treatment plant in Brazil after they had been treated by mesophilic anaerobic digestion. Quantitative polymerase chain reaction (PCR) was used for spiking experiments to compare the detection limits of feasible methods, such as beef extract elution and ultracentrifugation. Tests were performed to detect the inhibition levels and the bacteriophage PP7 was used as an internal control. The results showed that the inhibitors affected the efficiency of the PCR reaction and that beef extract elution is a suitable method for detecting enteric viruses, mainly AdV from biosolid samples. All of the viral groups were detected in the biosolid samples: AdV (90%), RVA, NoV GII (45%) and HAV (18%), indicating the viruses' resistance to the anaerobic treatment process. This is the first study in Brazil to detect the presence of RVA, AdV, NoV GII and HAV in anaerobically digested sludge, highlighting the importance of adequate waste management.

The shift from low to high non-structural protein 1 expression in rotavirus-infected MA-104 cells

A hallmark of group/species A rotavirus (RVA) replication in MA-104 cells is the logarithmic increase in viral mRNAs that occurs four-12 h post-infection. Viral protein synthesis typically lags closely behind mRNA synthesis but continues after mRNA levels plateau. However, RVA non-structural protein 1 (NSP1) is present at very low levels throughout viral replication despite showing robust protein synthesis. NSP1 has the contrasting properties of being susceptible to proteasomal degradation, but being stabilised against proteasomal degradation by viral proteins and/or viral mRNAs. We aimed to determine the kinetics of the accumulation and intracellular distribution of NSP1 in MA-104 cells infected with rhesus rotavirus (RRV). NSP1 preferentially localises to the perinuclear region of the cytoplasm of infected cells, forming abundant granules that are heterogeneous in size. Late in infection, large NSP1 granules predominate, coincident with a shift from low to high NSP1 expression levels. Our results indicate that rotavirus NSP1 is a late viral protein in MA-104 cells infected with RRV, presumably as a result of altered protein turnover.

Efficiency of the induced mating technique for Toxorhynchites theobaldi (Diptera, Culicidae)

Toxorhynchites mosquitoes play important ecological roles in aquatic microenvironments, and are frequently investigated as potential biological control agents of mosquito disease vectors. Establishment of Toxorhynchites laboratory colonies can be challenging because for some species, mating and insemination either do not occur or require a prohibitive amount of laboratory space for success. Consequently, artificial insemination techniques have been developed to assist with mass rearing of these species. Herein we describe an adapted protocol for colony establishment of T. theobaldi, a species with broad distribution in the Neotropics. The success of the technique and its implications are discussed.

Compositional and structural variabilities of Mg-rich iron oxide spinels from tuffite

Maghemite (γFe2O3) from tuffite is exceptionally rich in Mg, relatively to most of those reportedly found in other mafic lithosystems. To investigate in detail the compositional and structural variabilities of this natural magnetic iron oxide, sets of crystals were isolated from samples collected at different positions in a tuffite weathering mantle. These sets of crystal were individually powdered and studied by X-ray diffractometry, Mössbauer spectroscopy, magnetization measurements and chemical analysis. Lattice parameter of the cubic cell (a0) was found to vary from 0.834(1) to 0.8412(1) nm. Lower a0-values are characteristic of maghemite whereas higher ones are related to a magnetite precursor. FeO content ranges up to 17 mass % and spontaneous magnetization ranges from 8 to 32 J T-1 kg-1. Zero-field room temperature Mössbauer spectra are rather complex, indicating that the hyperfine field distributions due to Fe3+ and mixed valence Fe3+/2+ overlap. The structural variabilities of the (Mg, Ti)-rich iron oxide spinels is essentially related to the range of chemical composition of its precursor (Mg, Ti)-rich magnetite, and probably to the extent to which it has been oxidized during transformation in soil.

Structural quality of soils cultivated with coffee and pasture in an environmental protection area

Studies of soils in Environmental Protection Areas (EPAs) are of great importance, because they are an essential component of ecosystems, directly interfering in environmental sustainability. The objective of this study was to evaluate the structural quality of soil cultivated with coffee and used as pasture in the Capituva's River microbasin, which is located in the Environmental Protection Area in Coqueiral, south of the state of Minas Gerais. Uniaxial compression test (preconsolidation test) and soil resistance to penetration were used. Undisturbed samples were taken from the surface layer (0-5 cm) of the soils in the area: a typic dystrophic Red Latosol (LVd - Oxisol), a typic eutrophic Red Argisol (PVe - Ultisol), and a typic dystrophic Haplic Cambisol (CXbd - Inceptisol). A significant linear positive correlation was observed between the results of the preconsolidation test and soil resistance to penetration. Load bearing capacity of soil could be estimated accordingly by means of penetration resistance for LVd, PVe, and CXbd. Cambisol - CXbd showed lower loading support capacity and resistance to penetration than LVd and PVe, due to the better crop management in this soil that resulted in higher physical quality which accounts for higher production and environmental sustainability.

Diversity and efficiency of bradyrhizobium strains isolated from soil samples collected from around sesbania virgata roots using cowpea as trap species

The genetic diversity of ten Bradyrhizobium strains was evaluated for tolerance to high temperatures, to different salinity levels and for the efficiency of symbiosis with cowpea plants (Vigna unguiculata (L.) Walp.). Eight of these strains were isolated from nodules that appeared on cowpea after inoculation with suspensions of soil sampled from around the root system of Sesbania virgata (wand riverhemp) in ecosystems of South Minas Gerais. The other two strains used in our analyses as references, were from the Amazon and are currently recommended as cowpea inoculants. Genetic diversity was analyzed by amplifying repetitive DNA elements with the BOX primer, revealing high genetic diversity with each strain presenting a unique band profile. Leonard jar assays showed that the strains UFLA 03-30 and UFLA 03-38 had the highest N2-fixing potentials in symbiosis with cowpea. These strains had more shoot and nodule dry matter, more shoot N accumulation, and a higher relative efficiency than the strains recommended as inoculants. All strains grew in media of pH levels ranging from 4.0 to 9.0. The strains with the highest N2-fixing efficiencies in symbiosis with cowpea were also tolerant to the greatest number of antibiotics. However, these strains also had the lowest tolerance to high salt concentrations. All strains, with the exceptions of UFLA 03-84 and UFLA 03-37, tolerated temperatures of up to 40 ºC. The genetic and phenotypic characteristics of the eight strains isolated from soils of the same region were highly variable, as well as their symbiotic efficiencies, despite their common origin. This variability highlights the importance of including these tests in the selection of cowpea inoculant strains.

Rhizosphere properties of maize genotypes with contrasting phosphorus efficiency

An experiment was conducted in a growth chamber to evaluate characteristics of the rhizosphere of maize genotypes contrasting in P-use efficiency, by determining length and density of root hairs, the rhizosphere pH and the functional diversity of rhizosphere bacteria. A sample of a Red Oxisol was limed and fertilized with N, K and micronutrients. In the treatment with the highest P level, 174 mg kg-1 P was added. Each experimental unit corresponded to a PVC rhizobox filled with 2.2 dm-3 soil. The experiment was completely randomized with three replications in a 5 x 2 factorial design, corresponding to five genotypes (H1, H2 and H3 = P-efficient hybrids, H4 and H5 = P-inefficient hybrids) and two P levels (low = 3 mg dm-3, high = 29 mg dm-3). It was found that 18 days after transplanting, the nodal roots of the hybrids H3 and H2 had the longest root hairs. In general, the pH in the rhizosphere of the different genotypes was higher than in non-rhizosphere soil, irrespective of the P level. The pH was higher in the rhizosphere of lateral than of nodal roots. At low P levels, the pH variation of the hybrids H2, H4 and H5 was greater in rhizospheric than in non-rhizospheric soil. The functional microbial activity in the rhizosphere of the hybrids H3 and H5 was highest. At low soil P levels, the indices of microbial functional diversity were also higher. The microbial metabolic profile in the rhizosphere of hybrids H1, H2, H3, and H5 remained unaltered when the plants were grown at low P. The variations in the rhizosphere properties could not be related to patterns of P-use efficiency in the tested genotypes.

Structural changes in latosols of the cerrado region: I - relationships between soil physical properties and least limiting water range

The agricultural potential of Latosols of the Brazilian Cerrado region is high, but when intensively cultivated under inappropriate management systems, the porosity can be seriously reduced, leading to rapid soil degradation. Consequently, accelerated erosion and sedimentation of springs and creeks have been observed. Therefore, the objective of this study was to evaluate structural changes of Latosols in Rio Verde, Goiás, based on the Least Limiting Water Range (LLWR), and relationships between LLWR and other physical properties. Soil samples were collected from the B horizons of five oxidic Latosols representing the textural variability of the Latosols of the Cerrado biome. LLWR and other soil physical properties were determined at various soil compaction degrees induced by uniaxial compression. Soil compaction caused effects varying from enhanced plant growth due to higher water retention, to severe restriction of edaphic functions. Also, inverse relationships were observed between clay content and bulk density values (Bd) under different structural conditions. Bd values corresponding to critical soil macroporosity (BdcMAC) were more restrictive to a sustainable use of the studied Latosols than the critical Bd corresponding to LLWR (BdcLLWR). The high tolerable compression potential of these oxidic Latosols was related to the high aeration porosity associated to the granular structure.

Structural changes in latosols of the cerrado region: II - soil compressive behavior and modeling of additional compaction

Currently in Brazil, as in other parts of the world, the concern is great with the increase of degraded agricultural soil, which is mostly related to the occurrence of soil compaction. Although soil texture is recognized as a very important component in the soil compressive behaviors, there are few studies that quantify its influence on the structural changes of Latosols in the Brazilian Cerrado region. This study aimed to evaluate structural changes and the compressive behavior of Latosols in Rio Verde, Goiás, through the modeling of additional soil compaction. The study was carried out using five Latosols with very different textures, under different soil compaction levels. Water retention and soil compression curves, and bearing capacity models were determined from undisturbed samples collected on the B horizons. Results indicated that clayey and very clayey Latosols were more susceptible to compression than medium-textured soils. Soil compression curves at density values associate with edaphic functions were used to determine the beneficial pressure (σ b) , i.e., pressure with optimal water retention, and critical pressure (σcrMAC), i.e., pressure with macroporosity below critical levels. These pressure values were higher than the preconsolidation pressure (σp), and therefore characterized as additional compaction. Based on the compressive behavior of these Latosols, it can be concluded that the combined preconsolidation pressure, beneficial pressure and critical pressure allow a better understanding of compression processes of Latosols.

Nitrogen fertilizer use efficiency, recovery and leaching of an alexandergrass pasture

Nitrogen usually determines the productive potential of forage crops, although it is highly unstable in the environment. Studies on recovery rates and use efficiency are important for more reliable fertilizer recommendations to reduce costs and avoid environmental pollution. The purpose of this study was to evaluate N use efficiency and recovery rate of Alexandergrass pasture (Brachiaria - Syn. Urochloa plantaginea) as well as N-NO3- and N-NH4+ soil concentrations using different levels of N fertilization under two grazing intensities. The experiment was arranged in a randomized block design in a factorial scheme with three replications. Treatments consisted of three N rates (0, 200 and 400 kg ha-1 N) and two grazing intensities termed low mass (LM; forage mass of 2,000 kg ha-1 of DM) and high mass (HM; forage mass of 3,600 kg ha-1 of DM) under continuous stocking and variable stocking rates. Results of N fertilization with 200 kg ha-1 were better than with 400 kg ha-1 N. There was a significant effect of N rates on soil N-NO3-concentration with higher levels in the first layer of the soil profile in the treatment with 400 kg ha-1 N. Grazing intensity also affected soil N-NO3- concentration, by increasing the levels under the higher stocking rate (lower forage mass).

Chemical and structural characterization of soil humic substances under agroforestry and conventional systems

Studies have proven that the agroforestry systems in the semi-arid region of the State of Ceará, Brazil, induce an increase in soil organic C levels. Notwithstanding, there is no information if this increase also results in qualitative changes in different pools of soil organic matter. The objective of this study was to verify the possible chemical and structural alterations in fulvic and humic acids of a Luvisol in areas adopting agroforestry, traditional intensive cultivation and native forest in a long-term experiment conducted in the semi-arid region of Ceará State, Brazil. The study was conducted in an experimental area of the National Goat Research Center (Embrapa) in Sobral, CE. The following treatments were evaluated: agrosilvopasture (AGP), silvopasture (SILV), intensive cultivation under fallow (ICF), and areas with native forest (NF). Soil fulvic and humic acids fractions were extracted from the 0-6 and 6-12 cm layers and characterized by elemental composition, thermogravimetry and infrared spectroscopy analyses. The elemental composition analysis of humic acids confirmed the data found for fulvic acids, showing reduction in the C, H and N levels, followed by an increase in O contents in the AGP and ICF treatments over SILV and NF. In all treatments, except to SILV in the 0-6 cm layer, the percentage of mass loss was highest (300-600 °C) for humic acids in the thermally most stable region. Despite the similarity between infrared spectra, soil fulvic acids in the SILV treatment extracted from 6-12 cm depth decrease the absorption bands at 1708 and 1408 cm-1 followed by an increase in the absorption band at 1608 cm-1 attributed to aromatic C=C groups. This behavior suggests an increase in the aromatic character of the structure. The AGP and ICF treatments, which increase the soil tilling, favored the maintenance of humic substances with a more aromatic character in the soil than SILV and NF. The less aromatic humic substances in the SILV treatment resulted in an increase of exchange sites of soil organic matter, indicating improved nutrient cycling and maintenance of productivity in the system.

Assessment of Efficiency and Adequacy of Retention Terraces

Due to losses caused by water erosion, the development of techniques that increase the efficiency of soil conservation practices is fundamental. Terracing of agricultural lands is an important conservation practice. Bearing in mind that improperly built terraces may negatively affect the landscape, the purpose of this work was to evaluate the efficiency as well as the adequacy of retention terraces. Assessments were performed in four terraces implanted in different states, all located in the mideastern region of the state of Minas Gerais. The water storage efficiency of the terraces was determined by comparing the effective with the required storage capacity, as established in the project. Proposals were also made for the adequacy of the assessed terraces, based on the correction of the characteristics that jeopardized storage efficiency. The storage efficiency of three of the four assessed terraces was below the required levels (0.5-13 %). The main properties influencing storage capacity were: uniformity of ridge crest height, terrace end closure, and the cross section finishing. In two of the three low-efficiency terraces, the correction of these characteristics proved sufficient to raise the storage efficiency to nearly 100 %.

Structural quality of a no-tillage red latosol 50 months after gypsum aplication

Gypsum application may enhance the soil quality for plants in terms of soil chemical and physical properties. The objective of this study was to evaluate the effects of gypsum application on the structural quality of a no-tillage Red Latosol. The experiment was initiated in September 2005 in Guarapuava-PR, with gypsum applications of 0; 4; 8; and 12 Mg ha-1 on the soil surface. In November 2009, two soil blocks were sampled from the 0-0.3 m layer for visual evaluation of the soil structure quality (Sq) and to determine the aggregate-tensile strength (ATS). Soil penetration resistance (PR) and gravimetric moisture (H%) of the 0-0.300 m layer were evaluated, and soil cores were collected (layers 0.000-0.075 and 0.075-0.150 m), to determine soil bulk density (BD), total soil porosity (TP), microporosity (Mi), and macroporosity (Ma). Data were subjected to analysis of regression at 5 %. No significant effects of gypsum application on ATS and H % of aggregates were observed, but for Sq, a quadratic effect (0.000-0.075 m) and linear increase (0.075-0.150 and 0.150-0.300 m) were stated, indicating soil quality decrease, although Sq remained mostly below 3.0, with good to intermediate soil quality. Soil PR increased with gypsum, but also remained below critical levels. No effect was observed for soil H % at the moment of PR determination on the field. The gypsum applications decreased BD in the 0.075-0.150 m layer, and increased PT and Ma, while in 0.000-0.075 m some Ma was converted to Mi, without affecting PT and BD. These last results indicate a gain in soil structural quality by gypsum applications, but the higher scores of soil structure and values of soil penetration resistance, though still below thresholds, should be monitored to prevent limitations to soil use in the future.