Do water regimes affect iron‐plaque formation and microbial communities in the rhizosphere of paddy rice?

Pot experiments were conducted to investigate the effect of soil water regimes on the formation of iron (Fe) plaque on the root surface of rice seedlings (Oryza sativa L.) and on the microbial functional diversity in a paddy soil. The rice seedlings were subjected to three moisture regimes (submergence, 100%, and 60% water‐holding capacity [WHC]), and were grown for 5 and 11 weeks. Aerobic lithotrophic Fe(II)‐oxidizing (FeOB) and acetate‐utilizing Fe(III)‐reducing bacteria (FeRB) in the rhizosphere and non‐rhizosphere soil were determined at 5 weeks using the most probable number (MPN) method. The carbon substrate use patterns of the microbial communities in the rhizosphere and non‐rhizosphere soil samples were determined at 11 weeks using Biolog‐GN2 plates. The amount of Fe plaque (per unit dry root weight) was much higher under submerged conditions than at lower soil moisture contents and decreased with plant age. There was a positive correlation between the amount of Fe plaque and phosphorus accumulated in the Fe plaque at both sampling times (r = 0.98 and 0.92, respectively, n = 12). Numbers of FeOB and FeRB in the submerged soil were lower than in aerobic soil, but by two orders of magnitude higher in the rhizosphere than in the bulk soil. On the other hand, the functional diversity of the rhizosphere microbial communities was much higher than that of the non‐rhizosphere soil, irrespective of soil water regimes. We conclude that soil flooding results in a decreased number and diversity of Fe‐oxidizing/reducing bacteria, while increasing the Fe‐plaque formation.     

Do metal contamination and plant species affect microbial abundance and bacterial diversity in the rhizosphere of metallophytes growing in mining areas in a semiarid climate?

Journal of Soils and Sediments - Mining areas are low-quality habitats for macro- and microorganisms’ development, mainly due to the degradation of the soil quality by metal pollution. The...     

Do elevated soil concentrations of metals affect the diversity and activity of soil invertebrates in the long‐term?

This study aimed to elucidate the response of diversity and activity of soil invertebrates to elevated soil metal concentrations that were a result of sewage sludge application. Field sampling of soil invertebrates was carried out from 2002 to 2004 at an experimental site established in 1982 to test the effects on crop production of metal contamination from sewage sludge applications with elevated concentrations of zinc (Zn), copper (Cu) and nickel (Ni) with certain treatments exceeding the current UK statutory limits for the safe use of sludge on land. At metal concentrations within the limits, none of the invertebrates sampled showed adverse effects on their abundance or overall community diversity (from Shannon–Weiner index). At concentrations above the limits, individual taxa showed sensitivity to different metals, but overall diversity was not affected. Earthworm abundance was significantly reduced at total Cu concentrations at and above 176 mg kg^?1, while nematode and enchytraeid abundances were sensitive to Cu and high Zn concentrations. Correspondingly, litter decomposition was lower in Zn and Cu treatments although there was no direct relationship between decomposition and soil invertebrate abundance or diversity. Such enduring changes in both soil biodiversity and biological activity around the current UK regulatory limits warrant further investigation to determine whether they indicate detrimental damage to soil functioning over the long‐term.     

How does sonication affect the mineral and organic constituents of soil aggregates?—A review

Application of ultrasound to disperse soil aggregates has been critical in enabling researchers to separate and analyze aggregate building blocks that include organic and mineral particles as well as mineral associated organic matter. But the forces generated in the process may also alter the dispersion products and, thus, potentially interfere with the interpretation of experimental results. This review summarizes present knowledge on experimental conditions that may lead to physical damage and chemical modifications of aggregate building blocks. The energy level at which physical disintegration of organic particles could be detected was as low as 60 J mL^–1. Physical damage of sand‐ and silt‐sized mineral particles was observed to commence at energy levels exceeding 700 J cm^–3. No evidence was found for the disintegration of particles within the clay‐size fraction of soils even though studies analyzing pure minerals such as kaolinite revealed particle breakage after application of energy amounts > 12,000 J cm^–3. Here we outline a strategy to minimize artifacts such as physical damage of mineral or organic particles resulting from ultrasonication by adopting a stepwise dispersion protocol involving successively higher energy levels, accompanied by a sequential separation of organic and mineral compounds.     

Does the compositional change of soil organic matter in the rhizosphere and bulk soil of tea plants induced by tea polyphenols correlate with Pb bioavailability?

Purpose

Soil organic matter (SOM) plays a vital role in controlling metal bioavailability. However, the relationship between SOM and its fractions, including water-soluble substances (WSS), fulvic acid (FA), humic acid (HA), and soil microbial biomass (SMB), to metal bioavailability in plants has not been thoroughly investigated. This study examined the compositional change of SOM after tea polyphenols (TPs) were added to the soil and its correlation with Pb bioavailability.

Materials and methods

Ultisol samples were collected from Fuyang, spiked with two levels (0 and 300 mg kg^?1 DW) of Pb, and aged for 30 days. Four uniform seedlings were transplanted to each plastic pot, which were filled with 3 kg of air-dried soil. After successful transplantation, three levels (0, 300, and 600 mg kg^?1 DW) of TPs were amended as irrigation solution for the pots. The Pb concentrations in different tissues of the tea plants were determined after 6 months. SOM, WSS, FA, HA, and SMB were extracted and quantified using a Multi N/C Total Organic Carbon Analyser.

Results and discussion

Adding TPs to Pb-polluted soils alleviated Pb toxicity to microorganisms and increased SMB and the rhizosphere effect. The rhizosphere SOM was lower than bulk SOM in Pb-unspiked soils, while the opposite results were observed in Pb-spiked soils. A similar inconsistency for HA in the rhizosphere and bulk soil between Pb-unspiked and Pb-spiked soils might explain the difference in SOM. FA increased with the addition of TPs in both the rhizosphere and bulk soils, which might be the result of TP transformation. Positive correlations are present between the compositions of rhizosphere SOM and Pb in different tissues of the tea plant. SMB correlated negatively with Pb in young leaves and stems. Compared to rhizosphere soil, SOM components in bulk soil were less strongly correlated with Pb in tea plants.

Conclusions

Addition of TPs to soil changes the components of SOM and Pb bioavailability. SOM and its fractions, including WSS, FA, HA, and SMB, show a close relationship to Pb in different tissues of the tea plants.     

For how long does the quality and quantity of residues in the soil affect the carbon compartments and CO<Subscript>2</Subscript>-C emissions?

Purpose

The mineralization/immobilization of nutrients from the crop residues is correlated with the quality of the plant material and carbon compartments in the recalcitrant and labile soil fractions. The objective of this study was to correlate the quality and quantity of crop residues incubated in the soil with carbon compartments and CO₂-C emission, using multivariate analysis.

Materials and methods

The experiment was conducted in factorial 4?+?2?+?5 with three replicates, referring to three types of residues (control, sugarcane, Brachiaria, and soybean), and two contributions of the crop residues in constant rate, CR (10 Mg ha^?1 residue), and agronomic rate, AR (20, 8, and 5 Mg ha^?1 residue, respectively, for sugarcane, soybean, and Brachiaria), evaluated five times (1, 3, 6, 12, and 48 days after incubation). At each time, we determined the CO₂-C emission, nitrogen and organic carbon in the soil, and the residues. In addition, the microbial biomass and water-soluble, labile, and humic substance carbons fractionated into fulvic acids, humic acids and humin were quantified.

Results and discussion

Higher CO₂-C emissions occurred in the soil with added residue ranging from 0.5 to 1.1 g CO₂-C m^?2 h^?1 in the first 6 days of incubation, and there was a positive correlation with the less labile organic soil fractions as well as residue type. In the final period, after 12 days of soil incubation, there was a higher relation of CO₂-C emission with carbon humin. The sugarcane and soybean residue (20 Mg ha^?1) promoted higher CO₂-C emission and the reduction of carbon residue. The addition of residue contributed to an 82.32 % increase in the emission of CO₂-C, being more significant in the residue with higher nitrogen availability.

Conclusions

This study shows that the quality and quantity of residue added to soil affects the carbon sequestration and CO₂-C emission. In the first 6 days of incubation, there was a higher CO₂-C emission ratio which correlates with the less stable soil carbon compartments as well as residue. In the final period of incubation, there is no effect of quality and quantity of residue added to soil on the CO₂-C emission.

     

Variations in the soil microbial community composition of a tropical montane forest ecosystem: Does tree species matter?

We investigated tree species effects on the soil microbial community in the tropical montane forest on Mt. Kinabalu, in Malaysian Borneo. We investigated microbial composition (lipid profile) and soil physicochemical parameters (pH, moisture, total C, N and phenolics concentration) in top 5-cm soils underneath two conifers (Dacrycarpus imbricatus and Dacrydium gracilis) and three broad-leaves (Lithocarpus clementianus, Palaquium rioence and Tristaniopsis clementis). We found that the primary difference in microbial composition was between conifer versus broad-leaves. The abundance of specific microbial biomarker lipids correlated with soil pH, total C and N. We conclude that tree species have significant impacts on the soil microbial community through their effects on soil pH, total C and N.     

Does altered rainfall regime change pesticide effects in soil? A terrestrial model ecosystem study from Mediterranean Portugal on the effects of pyrimethanil to soil microbial communities under extremes in rainfall

In this century, agroecosystems are subjected to multiple global change stressors acting in concert such as alterations in rainfall regimes and pesticide use. Alterations in rainfall regimes, characterised by more extreme intra-annual rainfall regimes, have been forecasted for the Mediterranean region. At the same time, the use of pesticides continues to rise. Here, we report the responses of soil microbial community to a model pesticide, i.e., fungicide pyrimethanil (PYR) under altered rainfall regimes (i.e., drought and heavy rainfall) two and eight weeks after PYR application. We measured the functional responses as enzyme activities, potential nitrification and BIOLOG carbon substrate utilisation. We also characterised the soil bacterial communities using polymerase chain reaction–denaturing gradient gel electrophoresis (PCR–DGGE) method. After two weeks, enzyme activities were mainly responsive to PYR and kinetic parameters, calculated from BIOLOG carbon substrate utilisation, indicated interaction effects from PYR and rain treatments. Bacterial band richness increased with PYR treatment under normal rain and drought regimes, but bacterial band richness was higher at 1X than 5X PYR under heavy rainfall. Bacterial community structure was also different with the PYR and rainfall treatments. By week eight, PYR treated soils remained functionally different from untreated soils. Bacterial band richness was consistent across PYR treatment regardless of rain regime. However, the bacterial community structure remained significantly different among the PYR treatments under different rain regimes. We conclude that rainfall extremes can alter the effect of PYR on the soil microbial community structure without altering PYR effects on soil functions (measured as enzyme activities, potential nitrification and BIOLOG carbon substrate utilisation).     

Making knowledge and meaning in communities of practice: What role may science play? The case of sustainable soil management in England

Social learning is gaining popularity as a tool for understanding and designing interactions between experts and farming communities to enhance the uptake of sustainable and innovative farming practices. To date, the literature has mainly focused on the technical role scientists and researchers play in social learning, as sources of or co‐producers of knowledge. Social learning, however, implies a dynamic between the creation of knowledge (what can be done) and the creation of meaning (what is considered worth doing). This paper addresses this research gap by exploring the roles that “expert” actors and their narratives perform in meaning‐creation. I argue that a sustainable soil management community of practice is emerging in England and discuss the dynamics of farmer participation in this community. I further argue that members of this community use scientific “experts” and narratives to inspire, justify and legitimise sustainable soil management as a valid way of being a “good farmer.” This paper thus stresses the role that scientific or “expert” actors and narratives play in communities of practice as contributors to meaning‐creation inherent to social learning. How soil degradation will be addressed is as much a technical question, a question of what can be done, as a question of meaning, a question of what land managers consider worthwhile doing. The scientific community thus needs to work with the farming community not only to co‐produce technical solutions, but also to co‐produce shared visions of agrarian futures which put soils at their heart.     

Variation of soil microbial biomass C and hydrolytic enzyme activities in a rangeland ecosystem: are slope aspect and position effective?

This study was conducted to investigate the effects of slope aspect and position on microbial biomass C (MBC) and some hydrolytic enzyme activities involved in soil N, P, and S cycles in a rangeland ecosystem of west central Iran. Soil samples were collected from three slope positions (summit, backslope, and footslope) of contiguous north- and south-facing slopes. Results indicated higher silt and clay content, soil organic C (SOC), total N (TN), and C/N ratio on the north-facing slope. Furthermore, MBC, alkaline phosphomonoesterase (ALP), acid phosphomonoesterase (ACP), arylsulfatase (ARS), urease (URS), L-asparaginase (LAS), and L-glutaminase (LGL) activities were greater by 46.1, 65.9, 58.6, 59.6, 52.6, 62.8, and 65.7%, respectively, on the north-facing slope compared to the south-facing one. Higher ratios of enzyme activities to MBC were observed on the north-facing slope. In contrast, per cent of inorganic N and microbial quotient were greater on the south-facing slope. The activity of ALP, ACP, ARS along with SOC, TN, and MBC values decreased from summit to footslope. Overall, our findings indicate that north-facing slope and summit position support greater microbial biomass and hydrolytic diversity.     

Steam treatment of surface soil: how does it affect water-soluble organic matter, C mineralization, and bacterial community composition?

Steam soil disinfestation is efficiently used in the field for pre-planting pest control. Providing steam to the soil must have consequences, either beneficial or detrimental for the soil functioning. We set up a laboratory experiment to quantify the soil quality dynamics induced by this agricultural practice. In steamed and control soil, we monitored the size, the activity, and the genetic structure of the bacterial community in the top 2 cm soil every second day over a 10-day period after the treatment. We also characterized the bioavailable organic matter using fluorescence and ATR-FTIR spectroscopy. We showed that steaming induced the release of twice as much dissolved organic carbon in steamed soil as in the control soil. This extra carbon was much less fluorescent and contained a higher proportion of aliphatic compounds (alkyl chains, primary alcohols). After an initial drop in the bacterial community, we observed a tenfold increase in the number of bacteria, a flush in carbon mineralization, and genetic structure modification, which could be related to the newly released carbon. Steam treatment showed strong but quickly reversible impacts on the soil functioning, enabling farmers to sow approximately 1 week after treatment.     

Linking Ni and Cr concentrations to soil mineralogy: does it help to assess metal contamination when the natural background is high?

Purpose

In soils from serpentinitic areas the natural background of Ni and Cr is so high that the assessment of contamination by comparing metal concentrations with some fixed thresholds may give unreliable results. We therefore sought a quantitative relation between serpentines and Ni and Cr concentrations in uncontaminated soils, evaluated if the approach may help in establishing a baseline, and discussed if additional anthropogenic inputs of Ni and Cr can be realistically individuated in these areas.

Materials and methods

We analysed the total, acid-extractable and exchangeable concentrations of Ni and the total and acid-extractable concentrations of Cr in 66 soil horizons, belonging to 19 poorly developed and uncontaminated Alpine soils. The soils had different amounts of serpentines, depending on the abundance of these minerals in the parent material. We calculated an index of abundance of serpentines in the clay fraction by XRD and related total metal contents to the mineralogical index. We then tested the regressions on potentially contaminated soils, developed on the alluvial plain of the same watershed.

Results and discussion

We found extremely high total concentrations of Ni (up to 1,887 mg kg^–1) and Cr (up to 2,218 mg kg^–1) in the uncontaminated soils, but only a small proportion was extractable. Total Ni and Cr contents were significantly related to serpentine abundance (r ²?=?0.86 and 0.74, respectively). The regressions indicated that even small amounts of serpentines induced metal contents above 200 mg kg^–1, and the 95% confidence limits were 75 and 111 mg kg^–1 of Ni and Cr, respectively. When the regressions were tested on the potentially contaminated soils, a good estimate was obtained for Cr, while the Ni concentration was overestimated, probably because of some leaching of this element.

Conclusions

The concentrations of Ni and Cr that can be expected in soils because of the presence of small amounts of serpentines are comparable to the amounts accumulated in the soil because of diffuse contamination and potentially contaminated soils had metal concentrations falling in the range expected from the presence of natural sources. Only in the case of very severe contamination events, the identification of anthropogenic sources adding to the natural background would be feasible.     

How do environmental factors and different fertilizer strategies affect soil CO2 emission and carbon sequestration in the upland soils of southern China?

Upland soils have been identified as a major CO₂ source induced by human activities, such as fertilizer applications. The aim of this study is to identify the characteristics of soil CO₂ emission and carbon balance in cropland ecosystems after continuous fertilizer applications over decades. The measurements of soil surface CO₂ fluxes throughout the years of 2009 and 2010 were carried out based on a fertilization experiment (from 1990) in a double cropping system rotated with winter wheat (Triticum aestivum L.) and maize (Zea mays L.) in upland soil in southern China. Four treatments were chosen from the experiment for this study: no-fertilizer application (SR), nitrogen–phosphorus–potassium chemical fertilizers (NPK), NPK plus pig manure (NPKM) and pig manure alone (M). Results showed that the mean value of soil CO₂ fluxes from 08:00 to 10:00 am could represent its daily mean value in summer period (June–August) and that from 09:00 am to 12:00 pm for the rest season of a year. Soil temperature and moisture combined together could explain 70–83% of variations of CO₂ emission. Annual cumulative soil CO₂ fluxes in the treatments with manure applications (8.2 ± 0.8 and 11.0 ± 1.2 t C ha⁻¹ in 2009, and 7.9 ± 0.9 and 11.1 ± 1.2 t C ha⁻¹ in 2010 in NPKM and M, respectively) were significantly higher than those in the treatments with non-manure addition (2.5 ± 0.2 and 3.4 ± 0.2 t C ha⁻¹ in 2009, and 2.1 ± 0.2 and 3.7 ± 0.3 t C ha⁻¹ in 2010 in SR and NPK, respectively). However, the treatments with manure applications represented a carbon sink in the soil (carbon output/input ratio < 1.0), which demonstrated potential for carbon sequestration.     

How does the application of different nitrification inhibitors affect nitrous oxide emissions and nitrate leaching from cow urine in grazed pastures?

Nitrous oxide (N₂O) is a potent greenhouse gas, and nitrate () is a water contaminant. In grazed grassland, the major source of both leaching and N₂O emissions is nitrogen (N) deposited in animal excreta, particularly in the urine. The objective of this study was to determine the effectiveness of two nitrification inhibitors: (i) a solution of dicyandiamide (DCD) and (ii) a liquid formulation of 3,4‐dimethylpyrazole phosphate (DMPP) for reducing N₂O emissions and leaching from urine patch areas in two grazed pasture soils under different environmental conditions. In the Canterbury Templeton soil, the nitrification rate of ammonium from the animal urine applied at 1000 kg N/ha was significantly decreased by the application of DCD (10 kg/ha) and DMPP (5 kg/ha). N₂O emissions, measured over a 3‐month period, from dairy cow urine applied to the Canterbury Templeton soil were 1.14 kg N₂O‐N/ha, and this was reduced to 0.43 and 0.39 kg N₂O‐N/ha by DCD and the liquid DMPP, respectively. These are equivalent to 62–66% reductions in the total N₂O emissions. Nitrate leaching losses from dairy cow urine applied to the Waikato Horotiu soil lysimeters were reduced from 628.6 kg ‐N/ha to 400.6 and 451.5 kg ‐N/ha by the application of DCD (10 kg/ha) or DMPP (1 kg/ha), respectively. There was no significant difference between the DCD solution and the liquid DMPP in terms of their effectiveness in reducing N₂O emissions or leaching under the experimental conditions of this study. These results suggest that both the liquid formulations of DCD and DMPP have the potential to be used as nitrification inhibitors to reduce N₂O emissions and leaching in grazed pasture soils.     

Trace metal pollution in freshwater sediments of the world’s largest mercury mining district: sources,spatial distribution,and environmental implications

Purpose

The Almadén mining district has suffered long-term extraction activity, and this has left significant areas of decommissioned mining liabilities. Nowadays, the uncontrolled runoff and related erosion and transport of trace metal-enriched soils and sediments affect the whole freshwater ecosystem. The goal of this study was to distinguish geogenic from mining-related sources of trace metals in freshwater sediments, to understand their dispersion in the watershed, and, finally, to evaluate the potential environmental implications for future corrective plans.

Materials and methods

Freshwater surface sediment samples were collected from ten points along the main streams of the watershed (nine inside the mining district and one control point outside the district). Sediments were air dried and analyzed by different standard methods for pH, total major and trace element concentrations, total organic carbon, and grain size. In addition to the determination of the enrichment factor, a multi-statistical approach was applied involving discriminant analysis, Student’s t test, and Mann-Whitney U analysis.

Results and discussion

Sediments inside the district contained high levels of major and trace elements with respect to the control point. The predominance of fine fractions in these sediment samples appears to be one of the most important factors that affects trace metal concentrations. Among the trace elements, not only Hg but also As, Pb, and Zn are discriminative geochemical markers, thus allowing the identification of the different mining sources and their individual or combined impact throughout the district. Furthermore, the high enrichment factors obtained for As, Hg, Pb, and Zn with respect to the local background values highlight the persistent and severe impact from the decommissioned mines on the freshwater surficial sediments and their potential geoavailable risk for aquatic organisms.

Conclusions

The geochemistry of freshwater sediments alone demonstrates that different contamination sources are recognizable within the mining district and these can be related to the specific decommissioned mines. In addition, the discrete sources can be clearly distinguished on the basis of the statistical analysis of the geochemical data. Despite the closure of the mines, stream sediments are still the main repository of trace metals within the district, and they are therefore a potential threat to the freshwater ecosystem.

     

Homogeneous inoculation vs. microbial hot spots of isolated strain and microbial community: What is the most promising approach in remediating 1,2,4-TCB contaminated soils?

We investigated the effectiveness of different inoculation approaches in enhancing the mineralization of [U-¹⁴C] labeled 1,2,4-trichlorobenzene (1,2,4-TCB) in soil. Inoculation was conducted with a soil-borne 1,2,4-TCB mineralizing microbial community (MC) as well as the Bordetella sp. strain F2 originally isolated from this community as the key degrader organism (IS). Both were applied either via liquid medium (LM) or attached on clay particles (CP). Fluorescence in-situ hybridization in combination with ¹⁴C-1,2,4-TCB mineralization measurements as well as measurements of ¹⁴C-residues in soil were used to investigate the bioaugmentation efficiency of the different approaches. Bordetella sp. cell numbers increased about 2-5 times during the incubation process, indicating that the bacteria could survive and develop in the new soil habitat. While the native soil showed negligible 1,2,4-TCB mineralization rates, soil inoculated with the MC attached on CP showed the highest 1,2,4-TCB mineralization rate per Bordetella cell, whereas the other inoculum approaches showed an increased but lower contaminant mineralization. Additionally, the MC-CP approach showed the highest cumulative 1,2,4-TCB mineralization as well as the highest formation of bound ¹⁴C-residues which is most likely equivalent to ¹⁴C incorporated into the microbial biomass. Thus, our results allow the conclusion that the application of a specific microbe-clay-particle-complex is the most promising approach for an accelerated in-situ mineralization of chemicals in agricultural soils.     

Nitrogen pools and turnover in arable soils under different durations of organic farming: II: Source‐and‐sink function of the soil microbial biomass or competition with growing plants?

The following parameters were measured on seven field plots at 3 sites which had been under organic farming for different periods of time: mineral nitrogen (N _min) contents, in situ net nitrogen mineralization (N _net), soil microbial biomass carbon (C _mic), and nitrogen (N _mic) contents, and extractable organic N contents. The measurements were conducted every three weeks from spring 1995/1996 to autumn 1997. The objective was to test whether, under organic farming: 1) temporal fluctuations of N_mic contents over the course of the year are indicative for a source‐and‐sink function for plant‐available N of the soil microbial biomass, and 2) temporal variations in N_mic content can be related with in situ N_net or plant N uptake. N_min contents gradually increased after ploughing in autumn until late winter. During intensive plant growth in spring, values rapidly declined. In situ N_net fluctuated only moderately and reached high values during intensive plant growth (May—July) as well as after soil cultivation in autumn. The C_mic and N_mic contents generally were low in winter, increased in spring and reached maxima in late spring or summer. In spring, the increase in C_mic contents preceded the increase in N_mic contents, resulting in elevated C_mic:N_mic ratios until shooting of winter wheat. This corresponds to an uptake of available soil nitrogen by the plants at the expense of soil micro‐organisms. The subsequent increase in N_mic contents, coinciding with high plant N uptake rates, indicates an enhanced, plant‐induced N mobilization at that time. Possible mobilization mechanisms are discussed. Soil microbial biomass exerted a source‐and‐sink function for extractable organic N on some of the field plots. Estimates of in situ N_net measurements were neither correlated significantly with soil microbial biomass N, N_mic flux, N_mic turnover, nor with plant N uptake. Lower N_mic turnover rates on 41 years versus 3 years organically managed fields indicate a stabilizing effect of organic farming on soil microflora.