Seven years of enhanced water availability influences the physiological,structural, and functional attributes of a soil microbial community

Water availability is known to influence many aspects of microbial growth and physiology, but less is known about how complex soil microbial communities respond to changing water status. To understand how long-term enhancement of soil water availability (without flooding) influences microbial communities, we measured the seasonal dynamics of several community-level traits following >7 years of irrigation in a drought-prone tallgrass prairie soil. From late May to mid-September, water was supplied to the irrigated treatments based on calculated plant water demand. Phospholipid fatty acids (PLFA) were used to assess changes in microbial community structure and physiology. To assess the community-level physiological profile, microbial utilization of BIOLOG substrates was determined. After incubation for 2 days, the distribution of added ¹³C-glucose in microbial and respired pools was used as an index of substrate utilization efficiency. We also measured the relative contribution of fungi and bacteria to soil microbial biomass via substrate-induced respiration (SIR). Multivariate analysis of mol% PLFA and BIOLOG substrate utilization indicated that both water availability and sampling time influenced both the physiological and structural characteristics of the soil microbial community. Specific change in biomarker PLFA revealed a decreased ratio of cyclopropyl to ω7-precursors due to water addition, suggesting community-level stresses were reduced. Over the growing season, continuously greater water availability resulted in a 53% greater ratio of fungal to bacterial biomass using SIR, and a 65% increase in fungal PLFA. The number of substrates utilized by the cultivable microbial community tended to be greater in continuously wetted soil, especially during periods of low rainfall. While water dynamics appeared to be associated with some of the shifts in microbial community activity, structural and functional changes in the community appeared to be more closely linked to the cumulative effects of water regime on ecosystem properties. Seasonality strongly influenced microbial communities. The environmental factors associated with seasonal change need to be more closely probed to better understand the drivers of community structure and function.     

Multivariate approach to characterizing soil microbial communities in pristine and agricultural sites in Northwest Argentina

Land use effects on microbial communities may have profound impacts on agricultural productivity and ecosystem sustainability as they are critical in soil quality and health. The main aim of this study was to characterize the microbial communities of pristine and agricultural soils in the central Yungas region in Northwest Argentina. As a first step in the development of biological indicators of soil quality in this region, a comprehensive approach involving a structural and functional evaluation of microbial communities was used to detect changes in soil as consequence of land use. The sites selected included two pristine montane forest sites (MF1 and MF2), two plots under sugarcane monoculture for 40 and 100 years (SC40 and SC100), one plot under 20 years of soybean monoculture (SB20), a recently deforested and soybean cropped site (RC), and two reference sites of native forest adjacent to the sugarcane and soybean plots (PF1 and PF2). We used three microbial community profiling methods: denaturing gradient gel electrophoresis (DGGE) analysis of PCR amplified 16S rRNA genes, community-level physiological profiling (CLPP) using a BD oxygen biosensor system (BDOBS-CLPP) and phospholipid fatty acid (PLFA) analysis. Deforestation and agriculture caused expected increases in pH and decreases in organic carbon and microbial biomass. Additionally, shifts in the microbial community structure and physiology were detected with disturbance, including reduced diversity based on PLFA data. The higher respiratory response to several carbon substrates observed in agricultural soils suggested the presence of microbial communities with lower growth yield efficiency that could further reduce carbon storage in these soils.Using an integrated multivariate analysis of all data measured in this study we propose a minimum data set of variables (organic carbon, pH, sucrose and valeric acid utilizations, a17:0 and a15:0 PLFA biomarkers and the value of impact on microbial diversity) to be used for future studies of soil quality in Northwest Argentina.     

Soil Microbial Community Composition and Diversity in the Rhizosphere of a Chinese Medicinal Plant

Rehmannia glutinosa is an important medicinal plant, but there is a serious problem of decreasing productivity with its continuous cropping on the same land. We hypothesize some relationships between this problem and the disturbed soil ecosystem. In this work, two community‐based microbiological measurements, community‐level physiological profiling (CLPP) using Biolog sole carbon (C) source utilization tests and phospholipid ester–linked fatty acid (PLFA) profiles, were used to evaluate soil microbial community function and composition of different R. glutinosa cropping soils. Field investigation showed that the problems with continuous cropping occurred not only in 2‐year continuous fields but also in 5‐year rotation fields. Soil basal respiration and metabolic quotient were significantly greater in R. glutinosa cropping soils than in the noncropping controls. In contrast, the Shannon index from the Biolog data set was lower in R. glutinosa cropping soils. Both CLPP‐ and PLFA‐based principal component analyses (PCA) showed distinct groupings of soil microbial communities in R. glutinosa rhizosphere, and 11 PLFAs representing different microbes were identified from the principal component scores of PLFAs. Among these, an abundance of PLFA 18:2ω6,9, which is a biomarker of soil fungi, was significantly higher in R. glutinosa cropping soils than control soils. These results suggest an alteration of soil microbial community following R. glutinosa cropping, and this might be an important reason for the constraints associated with continuous cropping.     

核酸分析方法在土壤微生物多样性研究中的应用

土壤微生物多样性一般包括微生物分类群的多样性、遗传(基因)多样性、生态特征多样性和功能多样性。传统的分离培养方法和土壤微生物的生化研究手段具有一定的局限性,核酸分析方法为土壤微生物多样性研究注入了新活力。本文主要综述了近年来国内外研究土壤微生物多样性所采用的核酸提取方法及核酸分析方法。重点阐述了基于PCR的分子指纹技术、核酸杂交技术、基因芯片等核酸分析方法的原理、优缺点和应用。各种土壤微生物多样性研究方法的综合应用可扬长避短,起到相互补充的作用,从而能够提供更加丰富而准确的土壤微生物群落结构及种群丰度变化等方面的信息,也将成为这一领域今后的发展趋势。     

磷脂脂肪酸分析方法及其在土壤微生物多样性研究中的应用

磷脂脂肪酸（PLFA）是活体微生物细胞膜的重要组分，不同类群的微生物可通过不同的生化途径合成不同的PLFA。一些PLFA可作为分析微生物量和微生物群落结构变化的“生物标记”。在土壤微生物分析中，越来越多地采用了PLFA方法。本文介绍了表征微生物的一些PLFA、从土壤中提取PLFA的方法以及用GC-MS分析PLFA的原理。本文还将常用的研究微生物多样性的几种方法进行了比较；传统的分析土壤微生物群落的方法依赖于培养技术，只能培养和分离出一小部分微生物群落；Biolog方法可用于研究土壤微生物群落生理多样性，即可反映微生物群落如何利用各种碳源底物，但对快速生长和适合在Biolog实验条件下生长的小部分群落成员有强烈的选择性；核酸分析方法的主要缺点是不能对土壤微生物进行定量分析；而PLFA方法相对于上述几种方法有诸多优势。本文通过一些实例证明PLFA方法的特色或独到之处，也分析了其缺点。采用PLFA方法并结合其他方法有助于获取土壤微生物群落多样性的更多和更全面而完整的信息。     

Choice of methods for soil microbial community analysis: PLFA maximizes power compared to CLPP and PCR-based approaches

Polyphasic studies that used phospholipid fatty acid analysis (PLFA) in conjunction with community level physiological profiling (CLPP) or PCR-based molecular methods were analyzed in order to evaluate the power of each strategy to detect treatment effects on soil microbial community structure (MCS). We found no studies where CLPP or PCR-based methods differentiated treatments that were not also differentiated by PLFA. In 14 of 32 studies (44%), PLFA differentiated treatments that were not resolved by CLPP analysis. In 5 of 25 studies (20%), PLFA differentiated treatments that were not resolved by PCR-based methods. We discuss PLFA, CLPP, and PCR-based methods with respect to power to discriminate change in MCS versus potential for characterization of underlying population level changes.     

Effects of tree identity dominate over tree diversity on the soil microbial community structure

This study investigated the possible effects of tree species diversity and identity on the soil microbial community in a species-rich temperate broad-leaved forest. For the first time, we separated the effects of tree identity and tree species diversity on the link between above and belowground communities in a near-natural forest. We established 100 tree clusters consisting of each three tree individuals represented by beech (Fagus sylvatica L.), ash (Fraxinus excelsior L.), hornbeam (Carpinus betulus L.), maple (Acer pseudoplatanus L.), or lime (Tilia spec.) at two different sites in the Hainich National Park (Thuringia, Germany). The tree clusters included one, two or three species forming a diversity gradient. We investigated the microbial community structure, using phospholipid fatty acid (PLFA) profiles, in mineral soil samples (0–10 cm) collected in the centre of each cluster.The lowest total PLFA amounts were found in the pure beech clusters (79.0 ± 23.5 nmol g⁻¹ soil dw), the highest PLFA amounts existed in the pure ash clusters (287.3 ± 211.3 nmol g⁻¹ soil dw). Using principle components analyses (PCA) and redundancy analyses (RDA), we found only for the variables ‘relative proportion of beech trees’ and ‘living lime fine root tips associated with ectomycorrhiza’ a significant effect on the PLFA composition. The microbial community structure was mainly determined by abiotic environmental parameters such as soil pH or clay content. The different species richness levels in the clusters did not significantly differ in their total PLFA amounts and their PLFA composition. We observed a tendency that the PLFA profiles of the microbial communities in more tree species-rich clusters were less influenced by individual PLFAs (more homogenous) than those from species-poor clusters.We concluded that tree species identity and site conditions were more important factors determining the soil microbial community structure than tree species diversity per se.     

Short-term effects of defoliation on the soil microbial community associated with two contrasting Lolium perenne cultivars

Intra-species variation in response to defoliation and soil amendment has been largely neglected in terms of the soil microbial community (SMC). The influence of defoliation and soil fertiliser amendment on the structure of the SMC was assessed with two Lolium perenne cultivars contrasting in ability to accumulate storage reserves. Plant response to defoliation was cultivar specific and depended on the nutrient amendment of the soil. Results suggested a greater ability to alter plant biomass allocation in the low carbohydrate accumulating cultivar (S23) compared to the high carbohydrate cultivar (AberDove) when grown in improved (IMP), but not in unimproved (UNI), soil. Although differences in plant growth parameters were evident, no treatment effects were detected in the size of the active microbial biomass (total phospholipid fatty acid (PLFA) 313.8 nmol g⁻¹ soil±33.9) or proportions of PLFA signature groups. A lower average well colour development (AWCD) of Biolog sole carbon source utilisation profiles (SCSUPs) in defoliated (D) compared to non-defoliated (ND) treatments may be indicative of lower root exudation 1 week following defoliation, as a consequence of lower root non-structural carbohydrate (NSC) concentrations. Within the bacterial community the lower cyclopropyl-to-precursor ratio of PLFAs, and the trans/cis ratio of 16:1w7, in UNI relative to IMP soil treatments indicates lower physiological stress in UNI soils regardless of L. perenne cultivar. Discrimination of broad scale SMC structure, measured by PLFA analysis, revealed that soil treatment interacted strongly with cultivar and defoliation. In IMP soils the SMCs discriminated between cultivars while defoliation had little effect. Conversely, in UNI soils defoliation caused a common shift in the SMC associated with both cultivars, causing convergence of overall community structure. Separation of SMC structure along the primary canonical axis correlated most strongly (P<0.001) with root:shoot ratio (47.6%), confirming that differences in cultivar C-partitioning between treatments were influential in defining the rhizosphere microbial community.     

Soil microbial community composition and land use history in cultivated and grassland ecosystems of coastal California

Phospholipid ester-linked fatty acid (PLFA) profiles were used to evaluate soil microbial community composition for 9 land use types in two coastal valleys in California. These included irrigated and non-irrigated agricultural sites, non-native annual grasslands and relict, never-tilled or old field perennial grasslands. All 42 sites were on loams or sandy loams of similar soil taxa derived from granitic and alluvial material. We hypothesized that land use history and its associated management inputs and practices may produce a unique soil environment, for which microbes with specific environmental requirements may be selected and supported. We investigated the relationship between soil physical and chemical characteristics, management factors, and vegetation type with microbial community composition. Higher values of total soil C, N, and microbial biomass (total PLFA) and lower values of soil pH occurred in the grassland than cultivated soils. The correspondence analysis (CA) of the PLFA profiles and the canonical correspondence analysis (CCA) of PLFA profiles, soil characteristics, and site and management factors showed distinct groupings for land use types. A given land use type could thus be identified by soil microbial community composition as well as similar soil characteristics and management factors. Differences in soil microbial community composition were highly associated with total PLFA, a measure of soil microbial biomass, suggesting that labile soil organic matter affects microbial composition. Management inputs, such as fertilizer, herbicide, and irrigation, also were associated with the distinctive microbial community composition of the different cultivated land use types.     

Controls on soil nitrogen cycling and microbial community composition across land use and incubation temperature

We conducted a laboratory incubation of forest (Scots pine (Pinus sylvestris) or beech (Fagus sylvatica)), grassland (Trifolium repens/Lolium perenne) and arable (organic and conventional) soils at 5 and 25 °C. We aimed to clarify the mechanisms of short-term (2-weeks) nitrogen (N) cycling processes and microbial community composition in relation to dissolved organic carbon (DOC) and N (DON) availability and selected soil properties. N cycling was measured by ¹⁵N pool dilution and microbial community composition by denaturing gradient gel electrophoresis (DGGE), phospholipid fatty acid (PLFA) and community level physiological profiles (CLPP). Soil DOC increased in the order of arable<grassland<forest soil while DON and gross N fluxes increased in the order of forest<arable<grassland soil; land use had no affect on respiration rate. Soil DOC was lower, while respiration, DON and gross N fluxes were higher at 25 than 5 °C. Gross N fluxes, respiration and bacterial biomass were all positively correlated with each other. Gross N fluxes were positively correlated with pH and DON, and negatively correlated with organic matter, fungal biomass, DOC and DOC/DON ratio. Respiration rate was positively correlated with bacterial biomass, DON and DOC/DON ratio. Multiple linear modelling indicated that soil pH, organic matter, bacterial biomass, DON and DOC/DON ratio were important in predicting gross N mineralization. Incubation temperature, pH and total-C were important in predicting gross nitrification, while gross N mineralization, gross nitrification and pH were important in predicting gross N immobilization. Permutation multivariate analysis of variance indicated that DGGE, CLPP and PLFA profiles were all significantly (P<0.05) affected by land use and incubation temperature. Multivariate regressions indicated that incubation temperature, pH and organic matter content were important in predicting DGGE, CLPP and PLFA profiles. PLFA and CLPP were also related to DON, DOC, ammonium and nitrate contents. Canonical correlation analysis showed that PLFA and CLPP were related to differences in the rates of gross N mineralization, gross nitrification and soil respiration. Our study indicates that vegetation type and/or management practices which control soil pH and mediate dissolved organic matter availability were important predictors of gross N fluxes and microbial composition in this short-term experiment.     

Phospholipid fatty acid composition of a 2,4,6-trinitrotolune contaminated soil and an uncontaminated soil as affected by a humification remediation process

Analysis of phospholipid fatty acids (PLFAs) was performed to investigate effects of 2,4,6-trinitrotoluene (TNT) contamination and soil remediation on microbial biomass and community structure. A TNT-contaminated and an uncontaminated soil from a former ammunition plant were analysed before and after a humification/remediation process. TNT contamination reduced microbial biomass but indicated only minor differences in PLFA composition between the contaminated and uncontaminated soils. The humification process increased microbial biomass and altered soil PLFA patterns to a larger degree than did TNT contamination.     

Development of pollution-induced community tolerance is linked to structural and functional resilience of a soil bacterial community following a five-year field exposure to copper

Copper (Cu) is accumulating in agricultural soils worldwide creating concern for adverse impacts on soil microbial communities and associated ecosystem services. In order to evaluate the structural and functional resilience of soil microbial communities to increasing Cu levels, we compared a Cu-adapted and a corresponding non-adapted soil microbial community for their abilities to resist experimental Cu pollution. Laboratory soil microcosms were set-up with either High-Cu soil from Cu-amended field plots (63 g Cu m⁻²) or with Low-Cu control soil from the same five-year field experiment. Laboratory treatments consisted of Cu amendments in the presence or absence of pig manure. Microbial activities (soil respiration, substrate-induced respiration, [³H]leucine incorporation), bacterial community structure (terminal restriction fragment length polymorphism, T-RFLP), community-level physiological profiles, and pollution-induced bacterial community tolerance (PICT detected using the [³H]leucine incorporation technique) were monitored for 12 weeks. The High-Cu and Low-Cu soil microbial communities initially exhibited almost identical structure and function and could only be distinguished from each other by their differential Cu tolerance. Experimental Cu pollution inhibited microbial activities, affected bacterial community structure, and induced further bacterial community tolerance to Cu. However, Low-Cu and High-Cu soil microbial communities showed essentially identical responses. Manure amendment did not protect against Cu toxicity and slightly increased Cu bioavailability as measured by a Cu-specific whole-cell bacterial biosensor. Our results indicate convergence of bacterial community structure and function in the High-Cu and Low-Cu soils during the five-year field experiment. We conclude that soil bacterial communities can exhibit structural and functional resilience to a five-year Cu exposure by virtue of their ability to develop Cu tolerance without affecting overall community structure. The observed increased Cu tolerance may involve phenotypic adaptation or selection at the micro-diversity level, for example an increased proportion of Cu-resistant strains within each bacterial species, which go undetected by T-RFLP community fingerprinting. Finally, our results indicate that Cu-dissolved organic matter complexes contribute to microbial toxicity in manure-amended soils implying that free Cu may comprise a poor predictor of metal toxicity.     

Microbial and microfaunal community structure in cropping systems with genetically modified plants

Soils from field sites at Foulum (DK), Narbons (FR) and Varois (FR) planted with genetically modified maize expressing either the insecticidal Bacillus thuringiensis protein (Bt) or herbicide tolerance (HT), as described elsewhere in this volume, were analysed for nematodes, protozoa and microbial community structure. These analyses were mirrored in single-species testing and in mesocosm experiments, and were coordinated with field samples taken for microarthropods, enchytraeids and earthworms so allowing for cross-comparison and a better understanding of the results observed in the field. Over the first 2 years of the field experiments (in 2002 and 2003), the effect of Bt-maize was within the normal variation expected in these agricultural systems. Sampling in 2004 and 2005 was expanded to include the effects of tillage (i.e. reduced tillage versus conventional tillage) and also the use of HT-maize. Tillage had major effects regardless of soil type (Varois or Foulum), with reduced-tillage plots having a greater abundance of microfauna and a different microbial community structure (measured both by phospholipid fatty-acid analysis (PLFA) and by community-level physiological profiling (CLPP)) from conventionally tilled plots. Grass, as a contrasting cropping system to maize, also had an effect regardless of soil type and resulted in greater microfaunal abundance and an altered microbial community structure. Differences in crop management, which for the Bt-maize was removal of the insecticide used to control European corn borer and for HT-maize was a change in herbicide formulation, were only tested at single sites. There were differences in microbial community structure (CLPP but not PLFA) and sporadic increases in protozoan abundance under the Bt-crop management. The HT-maize cropping system, which covered a shorter period and only one site, showed little change from the conventional system other than an altered microbial community structure (as measured by PLFA only) at the final harvest. The Bt-trait had a minimal impact, with fewer amoebae at Foulum in May 2003, fewer nematodes at Foulum in May 2004 but more protozoa at Varois in October 2002 and an altered microbial community structure (PLFA) at Foulum in August 2005. These were not persistent effects and could not be distinguished from varietal effects. Based on the field evaluations of microfauna and microorganisms, we conclude that there were no soil ecological consequences for these communities associated with the use of Bt- or HT-maize in place of conventional varieties. Other land management options, such as tillage, crop type and pest management regime, had significantly larger effects on the biology of the soil than the type of maize grown.     

Structure and function of microbial communities in the rhizosphere of Scots pine after tree-felling

We evaluated changes occurring in the rhizosphere microbial communities of Scots pine (Pinus sylvestris L.) due to tree-felling and decrease of the photosynthetic C flow into the soil under field conditions over one growing season. Samples were taken from tree rhizospheres, freshly felled stump rhizospheres and bulk soil. We used culture dependent (CFU counts, community level physiological profiles, CLPPs) and independent methods (fluorogenic MUF-substrates, PLFA pattern and PCR-DGGE) to monitor the microbial communities in soil samples. The numbers of cultivable bacteria and amounts of phosphatase activity in the rhizosphere of trees were significantly higher compared with those in the bulk soil. The organic C consuming community measured by CLPP was stimulated directly after the tree-felling in stump rhizospheres; utilization of the disintegration components of cellulose, hemicellulose and chitin increased. Furthermore, bacterial and fungal biomass as well as chitin decomposers (CFU) increased in the stump rhizosphere. After 11 weeks of tree-felling the stump rhizosphere soluble PO₄-P and NH₄-N as well as amounts of total C and N began to resemble the concentrations measured in the bulk soil. However, the stump rhizosphere community structure detected by PLFA and PCR-DGGE still resembled that of the tree rhizosphere.     

Effects of heavy metal contamination and remediation on soil microbial communities in the vicinity of a zinc smelter as indicated by analysis of microbial community phospholipid fatty acid profiles

Heavy metal contamination in an area immediately surrounding a zinc smelter has resulted in destruction of over 485 hectares of forest. The elevated levels of heavy metals in these soils have had significant impacts on the population size and overall activity of the soil microbial communities. Remediation of these soils has resulted in increases in indicators of biological activity and viable population size, which suggest recovery of the microbial populations. Questions remain as to how the metal contamination and subsequent remediation at this site have impacted the population structure of the soil microbial communities. In the current study, microbial communities from this site were analyzed by the phospholipid fatty acid (PLFA) procedure. Principal component analysis of the PLFA profiles indicated that there were differences in the profiles for soils with different levels of metal contamination, and that soils with higher levels of metal contamination showed decreases in indicator PLFAs for mycorrhizal fungi, Gram-positive bacteria, fungi, and actinomycetes. PLFA profiles for remediated sites indicated that remediated soils showed increases in indicator PLFAs for fungi, actinomycetes, and Gram-positive bacteria, compared to unremediated metal contaminated soils. These data suggest a change in the population structure of the soil microbial communities resulting from metal contamination and a recovery of several microbial populations resulting from remediation.     

Microbial communities in forest floors under four tree species in coastal British Columbia

We compare forest floor microbial communities in pure plots of four tree species (Thuja plicata, Tsuga heterophylla, Pseudotsuga menziesii, and Picea sitchensis) replicated at three sites on Vancouver Island. Microbial communities were characterised through community level physiological profiles (CLPP), and profiling of phospholipid fatty acids (PLFA).Microbial communities from cedar forest floors had higher potential C utilisation than the other species. The F layer of the forest floor under cedar contained significantly higher bacterial biomass (PLFA) than the F layer under the other three tree species. There were differences in microbial communities among the three sites: Upper Klanawa had the highest bacterial biomass and potential C utilisation; this site also had the highest N availability in the forest floors. Forest floor H layers under hemlock and Douglas-fir contained greater biomass of Gram positive, Gram negative bacteria and actinomycetes than F layers based on PLFA, and H layers under spruce contained greater biomass of Gram negative bacteria than F layers. There were no significant differences in bacterial biomass between forest floor layers under cedar. Fungal biomass displayed opposite trends to bacteria and actinomycetes, being lowest in cedar forest floors, and highest in the F layer and at the site with lowest N availability. There were also differences in community composition among species and sites, with cedar forest floors having a much lower fungal:bacterial ratio than spruce, hemlock and Douglas-fir. The least fertile Sarita Lake site had a much greater fungal:bacterial ratio than the more fertile San Juan and Upper Klanawa sites. Forest floor layer had the greatest effect on microbial community structure and potential function, followed by site, and tree species. The similarity in trends among measures of N availability and microbial communities is further evidence that these techniques provide information on microbial communities that is relevant to N cycling processes in the forest floor.     

Experimentally induced effects of heavy metal on microbial activity and community structure of forest mor layers

This study compared the toxic effects of adding chromium (Cr), zinc (Zn), lead (Pb), molybdenum (Mo), nickel (Ni), and cadmium (Cd) at three dose levels to mor layer samples in laboratory experiments. Microbial activity in the form of soil respiration was monitored for 64 days. At the end of the experimental period, the composition of the soil microbial community structure was analysed by phospholipid fatty acid (PLFA) analysis. The metals added induced changes in the microbial community structure and affected respiration negatively, indicating toxicity. The microbial community structure (principal component analysis of the PLFA pattern) for all metals was significantly related to microbial activity (cumulative respiration), indicating intimate links between microbial community structure and activity. The most striking result in this study was that the shift in the microbial community because of metal stress was similar for all metals. Thus, the PLFA i16:0 increased most in relative abundance in metal-polluted soils, followed by other PLFAs indicative of Gram-positive bacteria (10Me16:0, 10Me17:0, 10Me18:0, a17:0 and br18:0). The PLFA 16:1ω5 was consistently negatively affected by metal stress, as were the PLFAs 18:1, 18:1ω7 and 19:1a. However, a significant separation between Cr- and Cd-polluted soils was observed in the response of the PLFA cy19:0, which decreased in abundance with Cr stress, and increased with Cd stress. Furthermore, the PLFA 18:2w6, indicating fungi, only increased with Cr and Zn stress. The effective doses of the metals, ranked with regard to background metal concentrations, decreased in the order: Zn > Cr > Pb > Mo > Ni > Cd. We concluded that interpretation of results of microbial activity from experiments of metal toxicity should include microbial structural patterns and background metal concentrations.     

Effects of Lumbricus terrestris, Allolobophora chlorotica and Eisenia fetida on microbial community dynamics in oil-contaminated soil

Oil spills are one of the most common types of soil pollution. Bioremediation has become an attractive alternative to physicochemical methods of remediation, where feasible. Earthworms have been shown to stimulate the degradation of petroleum hydrocarbons in soil, and it was hypothesized that the role of earthworms in remediation lies in the enhancement of an oil degrading microbial community. The aim of this study was to characterize microbial activity and community dynamics in oil-contaminated soil incubated with or without earthworms. Three earthworm species (Eisenia fetida, Allolobophora chlorotica and Lumbricus terrestris) were incubated in crude oil polluted soil (ca. 10,000 mg/kg total petroleum hydrocarbons (TPH)) and a reference soil for 28 d. Control treatments with manual mixing and/or cattle dung amendment were also included. In the oil-contaminated soil, respiration and concentration of microbial biomass was significantly enhanced by earthworm amendment, and TPH concentrations decreased significantly. These effects were less evident in treatments with A. chlorotica, possibly due to a difference in behavior, since individuals of this endogeic species were found in a state of inactivity (aestivation). Microbial community dynamics were described by phospholipid fatty acid (PLFA) analyses. After 28 d, similar shifts in the soil PLFA composition were observed in the oil-contaminated soil irrespective of worm species. Fungal:bacterial ratios were increased in the presence of worms, but also by addition of dung as a food source, indicating a non-specific effect of metabolizable substrates. In contrast, the fatty acids 17:1ω8 (=Δ9-heptadecenoic acid) and 20:4ω6c (arachidonic acid) were specifically stimulated by the presence of earthworms in the oil-contaminated soil. The results showed that earthworms can contribute positively to bioremediation of oil-contaminated soil, but that the effect may be species-dependent.     

Effects of sulfamethoxazole on soil microbial communities after adding substrate

The effect of the antibiotic sulfamethoxazole (SMX) on soil bacteria was studied using two methods (leucine incorporation and Biolog plates) of estimating pollution-induced community tolerance (PICT). SMX was added to an agricultural soil in a microcosm setup. The addition of different substrates (manure and alfalfa), and a non-amended soil, were also studied over 5 weeks. PICT measurements were validated by comparison with other measurements. Community structure was assessed using phospholipid fatty acid (PLFA) analysis and community-level physiological profiling (CLPP), and bacterial growth was estimated using leucine incorporation. Increased PICT was found at SMX concentrations of 20 and 500 mg SMX kg⁻¹ soil in samples containing manure and alfalfa, and at 500 mg SMX kg⁻¹ soil in non-amended soil (only concentration tested) using leucine incorporation. No effect was seen at 1 mg SMX kg⁻¹ soil. It was not necessary to add any substrate to increase the microbial activity in order to detect the effects of a bacteriostatic toxicant such as SMX when using measures based on bacterial growth. Direct inhibition of bacterial growth 2 days after SMX addition was correlated to PICT. No major changes in PICT due to SMX addition were found when using Biolog plates. However, there was a tendency towards increased PICT at the higher SMX concentrations in the manure-amended soil. Thus, different methods of detecting PICT have different sensitivities in detecting the toxic effects of SMX. The effects of substrate amendment were reflected by changes in the microbial community, estimated using both PLFA and CLPP. SMX was found to have a clear effect at the two highest levels of SMX in the manure- and alfalfa-amended soils, with an increase in fungal and a decrease in bacterial PLFAs. Little difference in the PLFA composition was found in the non-amended soil. CLPP was only affected at the highest SMX concentration. Although different variables showed different sensitivities to the effects of SMX, the results were consistent with an initial decrease in bacterial growth rates of sensitive species, which eventually transformed into more tolerant species, altering the community composition.