Expression of the Mycobacterium bovis P36 gene in Mycobacterium smegmatis and the baculovirus/insect cell system

In the present study we evaluated different systems for the expression of mycobacterial antigen P36 secreted by Mycobacterium bovis. P36 was detected by Western blot using a specific antiserum. The P36 gene was initially expressed in E. coli, under the control of the T7 promoter, but severe proteolysis prevented its purification. We then tried to express P36 in M. smegmatis and insect cells. For M. smegmatis, we used three different plasmid vectors differing in copy number and in the presence of a promoter for expression of heterologous proteins. P36 was detected in the cell extract and culture supernatant in both expression systems and was recognized by sera from M. bovis-infected cattle. To compare the expression level and compartmentalization, the MPB70 antigen was also expressed. The highest production was reached in insect cell supernatants. In conclusion, M. smegmatis and especially the baculovirus expression system are good choices for the production of proteins from pathogenic mycobacteria for the development of mycobacterial vaccines and diagnostic reagents.

Extracellular matrix molecules play diverse roles in the growth and guidance of central nervous system axons

Axon growth and guidance represent complex biological processes in which probably intervene diverse sets of molecular cues that allow for the appropriate wiring of the central nervous system (CNS). The extracellular matrix (ECM) represents a major contributor of molecular signals either diffusible or membrane-bound that may regulate different stages of neural development. Some of the brain ECM molecules form tridimensional structures (tunnels and boundaries) that appear during time- and space-regulated events, possibly playing relevant roles in the control of axon elongation and pathfinding. This short review focuses mainly on the recognized roles played by proteoglycans, laminin, fibronectin and tenascin in axonal development during ontogenesis.

A low concentration of ouabain (0.18 µg/kg) enhances hypertension in spontaneously hypertensive rats by inhibiting the Na+ pump and activating the renin-angiotensin system

We investigated the effects of low ouabain concentrations on systolic (SAP) and diastolic (DAP) arterial pressures and on pressor reactivity in 3-month-old male spontaneously hypertensive rats (SHR). Arterial blood pressure (BP) and pressor reactivity to phenylephrine (PHE) were investigated before and after 0.18 μg/kg ouabain administration (N = 6). The influence of hexamethonium (N = 6), canrenone (N = 6), enalapril (N = 6), and losartan (N = 6) on ouabain actions was evaluated. Ouabain increased BP (SAP: 137 ± 5.1 to 150 ± 4.7; DAP: 93.7 ± 7.7 to 116 ± 3.5 mmHg; P<0.05) but did not change PHE pressor reactivity. Hexamethonium reduced basal BP in control but not in ouabain-treated rats. However, hexamethonium + ouabain increased DAP sensitivity to PHE. Canrenone did not affect basal BP but blocked ouabain effects on SAP. However, after canrenone + ouabain administration, DAP pressor reactivity to PHE still increased. Enalapril and losartan reduced BP and abolished SAP and DAP responses to ouabain. Enalapril + ouabain reduced DAP reactivity to PHE, while losartan + ouabain reduced SAP and DAP reactivity to PHE. In conclusion, a small dose of ouabain administered to SHR increased BP without altering PHE pressor reactivity. Although the renin-angiotensin system (RAS), Na+ pump and autonomic reflexes are involved in the effects of ouabain on PHE reactivity, central mechanisms might blunt the actions of ouabain on PHE pressor reactivity. The effect of ouabain on SAP seems to depend on the inhibition of both Na+ pump and RAS, whereas the effect on DAP seems to depend only on RAS.

In vitro and in situ activation of the complement system by the fungus Lacazia loboi

Since there are no studies evaluating the participation of the complement system (CS) in Jorge Lobo's disease and its activity on the fungus Lacazia loboi, we carried out the present investigation. Fungal cells with a viability index of 48% were obtained from the footpads of BALB/c mice and incubated with a pool of inactivated serum from patients with the mycosis or with sterile saline for 30 min at 37 ºC. Next, the tubes were incubated for 2 h with a pool of noninactivated AB+ serum, inactivated serum, serum diluted in EGTA-MgCl2, and serum diluted in EDTA. The viability of L. loboi was evaluated and the fungal suspension was cytocentrifuged. The slides were submitted to immunofluorescence staining using human anti-C3 antibody. The results revealed that 98% of the fungi activated the CS by the alternative pathway and no significant difference in L. loboi viability was observed after CS activation. In parallel, frozen histological sections from 11 patients were analyzed regarding the presence of C3 and IgG by immunofluorescence staining. C3 and IgG deposits were observed in the fungal wall of 100% and 91% of the lesions evaluated, respectively. The results suggest that the CS and immunoglobulins may contribute to the defense mechanisms of the host against L. loboi.

Microbial biomass and soil chemical properties under different land use systems in northeastern Pará

The increase in agricultural production in the Brazilian Amazon region is mostly a result of the agricultural frontier expansion, into areas previously influenced by humans or of native vegetation. At the same time, burning is still used to clear areas in small-scale agricultural systems, leading to a loss of the soil productive capacity shortly after, forcing the opening of new areas. This study had the objective of evaluating the effect of soil preparation methods that involve plant residue shredding, left on the surface or incorporated to the soil, with or without chemical fertilization, on the soil chemical and biological properties. The experiment was conducted in 1995, in an experimental field of Yellow Latosol (Oxisol) of the Embrapa Amazônia Oriental, northeastern Pará (Brazil). The experiment was arranged in randomized blocks, in a 2x6 factorial design, with two management systems and six treatments evaluated twice. The management systems consisted of rice (Oriza sativa), followed by cowpea (Vigna unguiculata) with manioc (Manihot esculenta). In the first system the crops were planted in two consecutive cycles, followed by a three-year fallow period (natural regrowth); the second system consisted of one cultivation cycle and was left fallow for three years. The following treatments were applied to the secondary forest vegetation: slash and burn, fertilized with NPK (Q+NPK); slash and burn, without fertilizer NPK (Q-NPK); cutting and shredding, leaving the residues on the soil surface, fertilized with NPK (C+NPK); cutting and shredding, leaving residues on the soil surface, without fertilizer (C-NPK); cutting and shredding, with residue incorporation and fertilized with NPK (I+NPK); cutting and shredding, with residue incorporation and without NPK fertilizer (I-NPK). The soil was sampled in the rainier season (April 2006) and in the drier season (September 2006), in the 0-0.1 m layer. From each plot, 10 simple samples were collected in order to generate a composite sample. In the more intensive management system the contents of microbial C (Cmic) and microbial N (Nmic) were higher, while the C (Corg) level was higher in the less intensive system. The treatments with highest Cmic and Nmic levels were those with cutting, shredding and distribution of biomass on the soil surface. Under both management systems, the chemical characteristics were in ranges that classify the soil as little fertile, although P and K (in the rainy season) were higher in the less intensive management system.

Effect of management systems and cover crops on organic matter dynamics of soil under vegetables

Vegetable production in conservation tillage has increased in Brazil, with positive effects on the soil quality. Since management systems alter the quantity and quality of organic matter, this study evaluated the influence of different management systems and cover crops on the organic matter dynamics of a dystrophic Red Latosol under vegetables. The treatments consisted of the combination of three soil tillage systems: no-tillage (NT), reduced tillage (RT) and conventional tillage (CT) and of two cover crops: maize monoculture and maize-mucuna intercrop. Vegetables were grown in the winter and the cover crops in the summer for straw production. The experiment was arranged in a randomized block design with four replications. Soil samples were collected between the crop rows in three layers (0.0-0.05, 0.05-0.10, and 0.10-0.30 m) twice: in October, before planting cover crops for straw, and in July, during vegetable cultivation. The total organic carbon (TOC), microbial biomass carbon (MBC), oxidizable fractions, and the carbon fractions fulvic acid (C FA), humic acid (C HA) and humin (C HUM) were determined. The main changes in these properties occurred in the upper layers (0.0-0.05 and 0.05-0.10 m) where, in general, TOC levels were highest in NT with maize straw. The MBC levels were lowest in CT systems, indicating sensitivity to soil disturbance. Under mucuna, the levels of C HA were lower in RT than NT systems, while the C FA levels were lower in RT than CT. For vegetable production, the C HUM values were lowest in the 0.05-0.10 m layer under CT. With regard to the oxidizable fractions, the tillage systems differed only in the most labile C fractions, with higher levels in NT than CT in the 0.0-0.05 m layer in both summer and winter, with no differences between these systems in the other layers. The cabbage yield was not influenced by the soil management system, but benefited from the mulch production of the preceding maize-mucuna intercrop as cover plant.

Structural changes and degradation of Red Latosols under different management systems for 20 years

Soils are the foundation of terrestrial ecosystems and their role in food production is fundamental, although physical degradation has been observed in recent years, caused by different cultural practices that modify structures and consequently the functioning of soils. The objective of this study was to evaluate possible structural changes and degradation in an Oxisol under different managements for 20 years: no-tillage cultivation with and without crop rotation, perennial crop and conventional tillage, plus a forested area (reference). Initially, the crop profile was described and subsequently, 10 samples per management system and forest soil were collected to quantify soil organic matter, flocculation degree, bulk density, and macroporosity. The results indicated structural changes down to a soil depth of 50 cm, with predominance of structural units ∆μ (intermediate compaction level) under perennial crop and no-tillage crop rotation, and of structural units ∆ (compacted) under conventional tillage and no-tillage. The soil was increasingly degraded in the increasing order: forest => no-tillage crop rotation => perennial crop => no-tillage without crop rotation => conventional tillage. In all managements, the values of organic matter and macroporosity were always below and bulk density always above those of the reference area (forest) and, under no-tillage crop rotation and perennial crop, the flocculation degree was proportionally equal to that of the reference area.

Early Changes in Soil Metabolic Diversity and Bacterial Community Structure in Sugarcane under Two Harvest Management Systems

Preharvest burning is widely used in Brazil for sugarcane cropping. However, due to environmental restrictions, harvest without burning is becoming the predominant option. Consequently, changes in the microbial community are expected from crop residue accumulation on the soil surface, as well as alterations in soil metabolic diversity as of the first harvest. Because biological properties respond quickly and can be used to monitor environmental changes, we evaluated soil metabolic diversity and bacterial community structure after the first harvest under sugarcane management without burning compared to management with preharvest burning. Soil samples were collected under three sugarcane varieties (SP813250, SP801842 and RB72454) and two harvest management systems (without and with preharvest burning). Microbial biomass C (MBC), carbon (C) substrate utilization profiles, bacterial community structure (based on profiles of 16S rRNA gene amplicons), and soil chemical properties were determined. MBC was not different among the treatments. C-substrate utilization and metabolic diversity were lower in soil without burning, except for the evenness index of C-substrate utilization. Soil samples under the variety SP801842 showed the greatest changes in substrate utilization and metabolic diversity, but showed no differences in bacterial community structure, regardless of the harvest management system. In conclusion, combined analysis of soil chemical and microbiological data can detect early changes in microbial metabolic capacity and diversity, with lower values in management without burning. However, after the first harvest, there were no changes in the soil bacterial community structure detected by PCR-DGGE under the sugarcane variety SP801842. Therefore, the metabolic profile is a more sensitive indicator of early changes in the soil microbial community caused by the harvest management system.

Effect of soil management on the white grub population and damage in soybean

To evaluate the effect of soil management systems on population of white grubs, (Phyllophaga cuyabana Moser), and on its damage in soybean, experiments were set up under no-tillage and conventional tillage (one disk plow, and a leveling disk harrow) areas. Primary tillage equipment, used in other soil management systems, such as moldboard plow, disk plow, chisel plow and heavy duty disk harrow were also tested. Fluctuation of P. cuyabana population and the extent of its damage to soybean was similar under no-tillage and conventional tillage systems. Results comparing a range of primary tillage equipment showed that it affected soil insect populations differently, depending on the time during the season in which tillage was executed. Larval mortality could mostly be attributed to their exposure to adverse factors, soon after tillage, than to changes in soil conditions. Reduction of white grub population was more evident in plots managed by heavier equipment, such as the moldboard plow. Soil tillage could be one component within the soil pest management system in soybean, however, its use can not be generalized.

Effects of soil management systems on soil microbial activity, bulk density and chemical properties

The objective of this experiment was to study the effects of soil management systems on the bulk density, chemical soil properties, and on the soil microbial activity on a Latossolo Vermelho distrófico (Oxisol). Soil samples were collected from plots under the following management conditions: a) natural dense "cerrado" vegetation (savanna); b) degraded Brachiaria decumbens pasture, 20 years old; c) no-tillage treatment with annual crop sequence (bean, corn, soybean and dark-oat in continuous rotation), 8 years old; d) conventional tillage treatment with crop residues added to the soil, and annual crop sequence, 10 years old. The continuous use of no-tillage system resulted in an increase in microbial biomass and decrease in soil basal respiration, therefore displaying evident long-term effects on the increase of soil C content. The no-tillage system also provided an improvement in bulk density and chemical properties of the soil. Hence, the no-tillage management system could be an alternative for the conservation and maintenance of physical and chemical conditions and the productive potential of "cerrado" soils.

Energetic analysis of landrace varieties and hybrids of corn produced in different technological levels of management

The aim of this study was to analyze, under the energetic point of view, the cultivation of corn in three management systems (low, medium and high-tech), using two landrace varieties ('Argentino' and 'BR da Várzea'), a double hybrid cultivar (SHS 4080) and a simple hybrid (IAC 8333). Five performance indicators were used: energy efficiency, liquid cultural energy, cultural efficiency, energy balance and productive energy efficiency. From the perspective of family farming, it was verified the largest social importance of the systems under low and medium levels of technology, due to the increase employment capacity of rural labor. The liquid cultural energy and energy balance were more favorable for the system under high technological level, unlike cultural efficiency and productive energy efficiency, which were significantly higher for medium and low technological levels. The variety 'Argentino' showed lower productive energy efficiency. The variety 'BR da Várzea', on the other hand, presented the potential to generate energy as much as the hybrids. In general, the biggest sustainability in the corn crop was achieved when the management system under medium and lower levels of technology were used.

Population density and weed infestation in organic no-tillage corn cropping system under different soil covers

Currently, one of the biggest challenges faced by organic no-tillage farming is weed control. Thus, the use of cropping practices that help in the control of weeds is extremely important. The objective of this study was to evaluate population density and level of weed infestation in an organic no-tillage corn cropping system under different soil covers. The experiment was conducted in a randomized block design with six repetitions and five treatments, consisting of three soil covers in an organic no-tillage system, and an organic and a conventional system, both without soil cover. The treatments with soil cover used a grass species represented by the black oat, a leguminous species represented by the white lupine, and intercropping between both species. Corn was sown with spacing of 1.0 m between rows and 0.20 m between plants, using the commercial hybrid AG 1051. Infestation in corn was evaluated at stages V5 and V10, and weed density was evaluated at stage V5. The use of black oat straw alone or intercropped with white lupine, in the organic no-tillage corn cropping system, reduced the percentage of weed infestation and absolute weed density. Management-intensive systems and systems without soil cover showed higher relative densities for species Oxalis spp., Galinsoga quadriradiata and Stachys arvensis. The species Cyperus rotundus showed the highest relative density on organic no-tillage corn cropping systems. Black oat straw in the organic no-tillage cropping system limited the productive potential of corn.