Microbial and enzymatic activity in soil after organic composting

Microbial activity and biochemical properties are important indicators of the impact of organic composting on soil. The objective of this study was to evaluate some indicators of soil microbial and biochemical processes after application of compost (household waste). A Typic Acrustox, sampled at a depth of 10 cm under Cerrado biome vegetation, was evaluated in three treatments: control (soil without organic compost amendment) and soil with two doses of domestic organic compost (10 and 20 g kg-1 soil). The following properties were evaluated: released C (C-CO2): microbial respiration 15 days after incubation; microbial biomass C (MBC); total glucose (TG); metabolic quotient (qCO2); and enzyme activity of β-glucosidase and acid and alkaline phosphatase. The application of household compost, at doses of 10 and 20 g kg-1 Typic Acrustox, resulted in significant gains in microbial activity, organic C and C stock, as evidenced by increased MBC and TG levels. On the other hand, qCO2 decreases indicated greater microbial diversity and more efficient energy use. The addition of compost, particularly the 20 g kg-1 dose, strongly influenced the enzyme β-glucosidase and phosphatase (acid and alkaline). The β-glucosidase activity was significantly increased and acid phosphatase activity increased more than the alkaline. The ratio of β-glucosidase to MBC was greater in the control than in the composted treatments which suggests that there were more enzymes in the control than in the substrate or that the addition of compost induced a great MBC increase.

An improved method for measuring soil microbial activity by gas phase flow injection analysis

The rate of carbon dioxide production is commonly used as a measure of microbial activity in the soil. The traditional method of CO2 determination involves trapping CO2 in an alkali solution and then determining CO2 concentration indirectly by titration of the remaining alkali in the solution. This method is still commonly employed in laboratories throughout the world due to its relative simplicity and the fact that it does not require expensive, specific equipment. However, there are several drawbacks: the method is time-consuming, requires large amounts of chemicals and the consistency of results depends on the operator's skills. With this in mind, an improved method was developed to analyze CO2 captured in alkali traps, which is cheap and relatively simple, with a substantially shorter sample handling time and reproducibility equivalent to the traditional titration method. A comparison of the concentration values determined by gas phase flow injection analysis (GPFIA) and titration showed no significant difference (p > 0.05), but GPFIA has the advantage that only a tenth of the sample volume of the titration method is required. The GPFIA system does not require the purchase of new, costly equipment but the device was constructed from items commonly found in laboratories, with suggestions for alternative configurations for other detection units. Furthermore, GPFIA for CO2 analysis can be equally applied to samples obtained from either the headspace of microcosms or from a sampling chamber that allows CO2 to be released from alkali trapping solutions. The optimised GPFIA method was applied to analyse CO2 released from degrading hydrocarbons from a site contaminated by diesel spillage.

Industrial and urban organic wastes increase soil microbial activity and biomass

Microbial processes have been used as indicators of soil quality, due to the high sensitivity to small changes in management to evaluate, e.g., the impact of applying organic residues to the soil. In an experiment in a completely randomized factorial design 6 x 13 + 4, (pot without soil and residue or absolute control) the effect of following organic wastes was evaluated: pulp mill sludge, petrochemical complex sludge, municipal sewage sludge, dairy factory sewage sludge, waste from pulp industry and control (soil without organic waste) after 2, 4, 6, 12, 14, 20, 28, 36, 44, 60, 74, 86, and 98 days of incubation on some soil microbial properties, with four replications. The soil microbial activity was highly sensitive to the carbon/nitrogen ratio of the organic wastes. The amount of mineralized carbon was proportional to the quantity of soil-applied carbon. The average carbon dioxide emanating from the soil with pulp mill sludge, corresponding to soil basal respiration, was 0.141 mg C-CO2 100 g-1 soil h-1. This value is 6.4 times higher than in the control, resulting in a significant increase in the metabolic quotient from 0.005 in the control to 0.025 mg C-CO2 g-1 Cmic h-1 in the soil with pulp mill sludge. The metabolic quotient in the other treatments did not differ from the control (p < 0.01), demonstrating that these organic wastes cause no disturbance in the microbial community.

MICROBIAL CHARACTERISTICS OF SOILS UNDER AN INTEGRATED CROP-LIVESTOCK SYSTEM

Integrated crop-livestock systems (ICLs) are a viable strategy for the recovery and maintenance of soil characteristics. In the present study, an ICL experiment was conducted by the Instituto Agronômico do Paraná in the municipality of Xambre, Parana (PR), Brazil, to evaluate the effects of various grazing intensities. The objective of the present study was to quantify the levels of microbial biomass carbon (MBC) and soil enzymatic activity in an ICL of soybean (summer) and Brachiaria ruziziensis (winter), with B. ruziziensis subjected to various grazing intensities. Treatments consisted of varying pasture heights and grazing intensities (GI): 10, 20, 30, and 40 cm (GI-10, GI-20, GI-30, and GI-40, respectively) and a no grazing (NG) control. The microbial characteristics analysed were MBC, microbial respiration (MR), metabolic quotient (qCO2), the activities of acid phosphatase, β-glucosidase, arylsuphatase, and cellulase, and fluorescein diacetate (FDA) hydrolysis. Following the second grazing cycle, the GI-20 treatment (20-cm - moderate) grazing intensity) contained the highest MBC concentrations and lowest qCO2 concentrations. Following the second soybean cycle, the treatment with the highest grazing intensity (GI-10) contained the lowest MBC concentration. Soil MBC concentrations in the pasture were favoured by the introduction of animals to the system. High grazing intensity (10-cm pasture height) during the pasture cycle may cause a decrease in soil MBC and have a negative effect on the microbial biomass during the succeeding crop. Of all the enzymes analyzed, only arylsuphatase and cellulase activities were altered by ICL management, with differences between the moderate grazing intensity (GI-20) and no grazing (NG) treatments.

Síntese de regioisômeros quirais a partir de D-manitol: obtenção de uma mistura de álcoois acetilênicos

The synthesis of chiral acetylenic regioisomers was described by using an appropriate intermediate such as isopropylidene glycerol, a synthon widely used in the enantioselective syntheses. This intermediate was prepared from D-mannitol. The nine obtained compounds have been characterized by their respective spectral data. The mixture of chiral acetylenic alcohols showed activity against Escherichia coli when tested through the monitoring of CO2 released during microbial respiration by using a conductimetric system.

Effect of compaction on microbial activity and carbon and nitrogen transformations in two oxisols with different mineralogy

The use of machinery in agricultural and forest management activities frequently increases soil compaction, resulting in greater soil density and microporosity, which in turn reduces hydraulic conductivity and O2 and CO2 diffusion rates, among other negative effects. Thus, soil compaction has the potential to affect soil microbial activity and the processes involved in organic matter decomposition and nutrient cycling. This study was carried out under controlled conditions to evaluate the effect of soil compaction on microbial activity and carbon (C) and nitrogen (N) mineralization. Two Oxisols with different mineralogy were utilized: a clayey oxidic-gibbsitic Typic Acrustox and a clayey kaolinitic Xantic Haplustox (Latossolo Vermelho-Amarelo ácrico - LVA, and Latossolo Amarelo distrófico - LA, respectively, in the Brazil Soil Classification System). Eight treatments (compaction levels) were assessed for each soil type in a complete block design, with six repetitions. The experimental unit consisted of PVC rings (height 6 cm, internal diameter 4.55 cm, volume 97.6 cm³). The PVC rings were filled with enough soil mass to reach a final density of 1.05 and 1.10 kg dm-3, respectively, in the LVA and LA. Then the soil samples were wetted (0.20 kg kg-1 = 80 % of field capacity) and compacted by a hydraulic press at pressures of 0, 60, 120, 240, 360, 540, 720 and 900 kPa. After soil compression the new bulk density was calculated according to the new volume occupied by the soil. Subsequently each PVC ring was placed within a 1 L plastic pot which was then tightly closed. The soils were incubated under aerobic conditions for 35 days and the basal respiration rate (CO2-C production) was estimated in the last two weeks. After the incubation period, the following soil chemical and microbiological properties were detremined: soil microbial biomass C (C MIC), total soil organic C (TOC), total N, and mineral N (NH4+-N and NO3--N). After that, mineral N, organic N and the rate of net N mineralization was calculated. Soil compaction increased NH4+-N and net N mineralization in both, LVA and LA, and NO3--N in the LVA; diminished the rate of TOC loss in both soils and the concentration of NO3--N in the LA and CO2-C in the LVA. It also decreased the C MIC at higher compaction levels in the LA. Thus, soil compaction decreases the TOC turnover probably due to increased physical protection of soil organic matter and lower aerobic microbial activity. Therefore, it is possible to conclude that under controlled conditions, the oxidic-gibbsitic Oxisol (LVA) was more susceptible to the effects of high compaction than the kaolinitic (LA) as far as organic matter cycling is concerned; and compaction pressures above 540 kPa reduced the total and organic nitrogen in the kaolinitic soil (LA), which was attributed to gaseous N losses.

Microbial and soil properties in restoration areas in the jequitinhonha valley, Minas Gerais

To mitigate the impacts of eucalypt monoculture, forestry companies in the Upper Jequitinhonha Valley (MG) have adopted the insertion of strips of native vegetation in-between the commercial plantations. The method used for the creation of these corridors is to allow spontaneous regrowth of native vegetation in areas previously under eucalypt. The objective of this study was to evaluate the effect of cover crops on microbial and soil properties for a detailed description of the restoration process of native vegetation in forest soils of the Jequitinhonha Valley. The treatments were represented by an initial restoration stage (< 4 years) with or without remaining eucalypt and the advanced restoration stage (> 4 years) with or without remaining eucalypt, plus the three controls: commercial eucalypt plantation, Cerrado vegetation and native forest. Soil samples were collected for three consecutive years in the dry and rainy season (August and February, respectively). The microbial activity, regardless of the presence of remaining eucalypt , did not differ among the restoration areas, except for the metabolic quotient (qCO2) in the rainy season of February 2007. At this time, this microbial activity was higher in the advanced restoration stage without eucalypt than initial restoration without eucalypt and advanced restoration with eucalypt. The restoration areas, in general, did not differ from the control: eucalypt plantation and Cerrado either. Compared to the forest, the levels of organic C, microbial C, basal respiration (Rbasal) and hydrolysis of fluorescein diacetate (FDA) in the restoration areas were, in general, lower and did not differ in qCO2 and microbial quotient (qMIC). In general, the soil quality was similar in the initial and advanced restoration stages. Most of the soil and microbial properties in the three years indicated that the restoration areas were most similar to the Cerrado. In the advanced restoration areas without eucalypt compared to Cerrado, the lower Rbasal in the 3rd year and the lower FDA and qMIC and higher qCO2 in the 2nd year indicated that the removal of the remaining eucalypt trees was unfavorable for restoration.

Microbial biomass and activity in litter during the initial development of pure and mixed plantations of Eucalyptus grandis and Acacia mangium

Studies on microbial activity and biomass in forestry plantations often overlook the role of litter, typically focusing instead on soil nutrient contents to explain plant and microorganism development. However, since the litter is a significant source of recycled nutrients that affect nutrient dynamics in the soil, litter composition may be more strongly correlated with forest growth and development than soil nutrient contents. This study aimed to test this hypothesis by examining correlations between soil C, N, and P; litter C, N, P, lignin content, and polyphenol content; and microbial biomass and activity in pure and mixed second-rotation plantations of Eucalyptus grandis and Acacia mangium before and after senescent leaf drop. The numbers of cultivable fungi and bacteria were also estimated. All properties were correlated with litter C, N, P, lignin and polyphenols, and with soil C and N. We found higher microbial activity (CO2 evolution) in litter than in soil. In the E. grandis monoculture before senescent leaf drop, microbial biomass C was 46 % higher in litter than in soil. After leaf drop, this difference decreased to 16 %. In A. mangium plantations, however, microbial biomass C was lower in litter than in soil both before and after leaf drop. Microbial biomass N of litter was approximately 94 % greater than that of the soil in summer and winter in all plantations. The number of cultivable fungi and bacteria increased after leaf drop, especially so in the litter. Fungi were also more abundant in the E. grandis litter. In general, the A. mangium monoculture was associated with higher levels of litter lignin and N, especially after leaf drop. In contrast, the polyphenol and C levels in E. grandis monoculture litter were higher after leaf drop. These properties were negatively correlated with total soil C and N. Litter in the mixed stands had lower C:N and C:P ratios and higher N, P, and C levels in the microbial biomass. This suggests more effective nutrient cycling in mixed plantations in the long term, greater stimulation of microbial activity in litter and soil, and a more sustainable system in general.

Relationships between microbial activity and soil physical and chemical properties in native and reforested Araucaria angustifolia forests in the state of São Paulo, Brazil

Araucaria angustifolia (Bert.) O. Kuntze is the main component of the Mixed Ombrophilous forest and, in the State of São Paulo, it is associated with a high diversity of soil organisms, essential for the maintenance of soil quality, making the conservation of this ecosystem a major and pressing challenge. The objective of this study was to identify the physical and chemical properties that are most closely correlated with dehydrogenase enzyme activity, basal respiration and microbial biomass under native (NF) and replanted (RF) Araucaria angustifolia forests in three regions of the state of São Paulo, in winter and summer. The main differentiating factors between the areas were also determined. Each forest was represented by three true replications; at each site, from around the araucaria trees, 15 soil samples (0-20 cm) were collected to evaluate the soil physical, chemical and microbiological properties. At the same points, forest litter was sampled to assess mass and chemical properties. The following microbiological properties were evaluated: microbial biomass carbon (MBC), basal respiration (CO2-C), metabolic quotient (Q: CO2), dehydrogenase enzyme activity (DHA) as well as the physical properties (moisture, bulk density, macroporosity and total porosity), soil chemical properties [pH, organic carbon (org-C), P, Ca, K, Mg, Al, H+Al], litter dry mass, and C, N and S contents. The data were subjected to analysis of variance (TWO-WAY: ANOVA). A Canonical Discriminant Analysis (CDA) and a Canonical Correlation Analysis (CCA) were also performed. In the soil under NF, the values of K, P, soil macroporosity, and litter dry mass were higher and Q: CO2 and DHA lower, regardless of the sampling period, and DHA was lower in winter. In the RF areas, the levels of moisture, porosity and Q: CO2 were higher in both sampling periods, and DHA was higher in winter. The MBC was only higher under NF in the summer, while the litter contents of C, N and S were greater in winter. In winter, CCA showed a high correlation of DHA with CO2-C, pH and H+Al, while in the summer org-C, moisture, Mg, pH and litter C were more associated with DHA and CO2-C. The CDA indicated H+Al, available P, total porosity, litter S content, and soil moisture as the most discriminating variables between NF and RF, but moisture was the most relevant, in both seasons and CO2-C only in winter. The combined analysis of CCA and CDA showed that the contribution of the microbiological variables to a differentiation of the areas was small at both samplings, which may indicate that the period after reforestation was long enough to allow an almost complete recovery of the microbial activity.

EFFECT OF WATER AVAILABILITY ON SOIL MICROBIAL BIOMASS IN SECONDARY FOREST IN EASTERN AMAZONIA

Soil microbial biomass (SMB) plays an important role in nutrient cycling in agroecosystems, and is limited by several factors, such as soil water availability. This study assessed the effects of soil water availability on microbial biomass and its variation over time in the Latossolo Amarelo concrecionário of a secondary forest in eastern Amazonia. The fumigation-extraction method was used to estimate the soil microbial biomass carbon and nitrogen content (SMBC and SMBN). An adaptation of the fumigation-incubation method was used to determine basal respiration (CO2-SMB). The metabolic quotient (qCO2) and ratio of microbial carbon:organic carbon (CMIC:CORG) were calculated based on those results. Soil moisture was generally significantly lower during the dry season and in the control plots. Irrigation raised soil moisture to levels close to those observed during the rainy season, but had no significant effect on SMB. The variables did not vary on a seasonal basis, except for the microbial C/N ratio that suggested the occurrence of seasonal shifts in the structure of the microbial community.

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.

Dinámica microbial del suelo asociada a diferentes estrategias de manejo de Phytophthora cinnamomi Rands en aguacate

La marchitez del aguacate es la enfermedad más limitante de este cultivo, cuyo agente causal más relevante es el oomycete Phytophthora cinnamomi Rands. Es por esto que se han desarrollado diferentes estrategias para su manejo integrado, pero aún prevalece el uso de productos químicos, como única medida de manejo, generando impactos negativos en el ambiente y la salud. Uno de los efectos perjudiciales que se ocasiona es la alteración de las poblaciones microbianas en el suelo. Este trabajo estuvo encaminado a conocer la dinámica microbiana del suelo, bajo diferentes estrategias de manejo de esta enfermedad, para lo cual se midió su dinamismo mediante unidades formadoras de colonias (UFC), para hongos, bacterias y actinomicetos, a partir de muestras de suelo y rizósfera de la raíz, bajo incubación en condiciones de anaerobiosis y aerobiosis, además se midió la actividad microbiana total, en condiciones de laboratorio, como complemento se cuantificaron microorganismos como: Trichiderma spp, bacterias formadoras de endosporas (BAFE), celulolíticos, proteolíticos, amilolíticos, solubilizadores de fosfato, fijadores asimbióticos de nitrógeno y promotores del crecimiento, como Pseudomonas spp., fluorescentes. Los resultados encontrados en esta investigación, sugieren que el uso individual y combinado de mantillo orgánico, material compostado de estiércol bovino, enmienda mineral y cascarilla de arroz y la propuesta de integración; incrementan significativamente la población y actividad microbiana aerobia, en la cual se identificaron microorganismos antagonistas como, Trichiderma spp., celulolíticos, Pseudomonas spp. fluorescentes y BAFE.

New Strategies on Molecular Biology Applied to Microbial Systematics

Systematics is the study of diversity of the organisms and their relationships comprising classification, nomenclature and identification. The term classification or taxonomy means the arrangement of the organisms in groups (rate) and the nomenclature is the attribution of correct international scientific names to organisms and identification is the inclusion of unknown strains in groups derived from classification. Therefore, classification for a stable nomenclature and a perfect identification are required previously. The beginning of the new bacterial systematics era can be remembered by the introduction and application of new taxonomic concepts and techniques, from the 50’s and 60’s. Important progress were achieved using numerical taxonomy and molecular taxonomy. Molecular taxonomy, brought into effect after the emergence of the Molecular Biology resources, provided knowledge that comprises systematics of bacteria, in which occurs great evolutionary interest, or where is observed the necessity of eliminating any environmental interference. When you study the composition and disposition of nucleotides in certain portions of the genetic material, you study searching their genome, much less susceptible to environmental alterations than proteins, codified based on it. In the molecular taxonomy, you can research both DNA and RNA, and the main techniques that have been used in the systematics comprise the build of restriction maps, DNA-DNA hybridization, DNA-RNA hybridization, sequencing of DNA sequencing of sub-units 16S and 23S of rRNA, RAPD, RFLP, PFGE etc. Techniques such as base sequencing, though they are extremely sensible and greatly precise, are relatively onerous and impracticable to the great majority of the bacterial taxonomy laboratories. Several specialized techniques have been applied to taxonomic studies of microorganisms. In the last years, these have included preliminary electrophoretic analysis of soluble proteins and isoenzymes, and subsequently determination of deoxyribonucleic acid base composition and assessment of base sequence homology by means of DNA-RNA hybrid experiments beside others. These various techniques, as expected, have generally indicated a lack of taxonomic information in microbial systematics. There are numberless techniques and methodologies that make bacteria identification and classification study possible, part of them described here, allowing establish different degrees of subspecific and interspecific similarity through phenetic-genetic polymorphism analysis. However, was pointed out the necessity of using more than one technique for better establish similarity degrees within microorganisms. Obtaining data resulting from application of a sole technique isolatedly may not provide significant information from Bacterial Systematics viewpoint

Immunomodulation by microbial ribosomes

Over the past twenty years, many authors have reported evidence of the immunoprotective capacity of ribosomes isolated from bacteria, fungi and parasites. Since 1971 we have explored the protective capacity of ribosomes isolated from a large variety of microorganisms responsible for human and animal diseases. More recently, using monoclonal antibodies raised against ribosomes and then selected for their ability to confer passive immunity to mice, we have studied the mechanism of the protection induced by ribosomes. These studies, in parallel with the development of a technology for the large scale production of ribosomes, have allowed us to achieve a new regard for ribosomal vaccines for use in human. The general concept of ribosomal vaccines in presented and examples of two such vaccines are described with data on the specific protection that they induce in mice against experimental infections with Klebsiella peneumoniae, Streptococcus pneumoniae, S. pyogenes and Haemophilus influenzae for the first one, and against Candida albicans type A and type B for the second one. Because of their high immunogenicity and their innocuity these vaccines represent a decisive improvement over classical microbial vaccines.