Impact of allelochemical exuded from allelopathic rice on soil microbial community

Allelopathic rice releases allelochemicals from its roots to paddy soils at early growth stages to inhibit neighboring weeds. However, little is currently known about the effects of allelochemicals on soil microbes. In this study, we show that allelopathic rice can have great impact on the population and community structure of soil microbes. Allelopathic rice PI312777 seedlings reduced the culturable microbial population and total PLFA when compared to non-allelopathic rice Liaojing-9. Similar results were observed when, instead of growing seedlings, soils were incubated with plant root exudates. This result demonstrates that the composition of root exudates from the rice varieties tested contributes to the soil microbial community. Further experiments showed that the microbial community was affected by the allelochemical 5,4′-dihydroxy-3′,5′-dimethoxy-7-O-β-glucopyranosylflavone exuded from allelopathic rice roots, through immediately hydrolyzing glucose with stimulation on soil bacteria and aglycone (5,7,4′-trihydroxy-3′,5′-dimethoxyflavone) with inhibition on soil fungi. This result indicates that the flavone O-glycoside can provide carbon and interact with soil microbes. PC analysis of the fatty acid data clearly separated the allelopathic PI312777 and the non-allelopathic Liaojing-9 variety (PC1 = 46.4%, PC2 = 20.3%). Similarly, the first principal component (PC1 = 37.4%) together with the second principal component (PC2 = 17.3%) explained 54.7% of the variation between the allelopathic and non-allelopathic root exudates. Furthermore, the canonical correlation between allelopathic root exudates and the flavone O-glycoside was statistically significant (Canonical R = 0.889, χ² (25) = 69.72, p = 0.0041). Although the data generated in this study were not completely consistent between culturable microbes and PLFA profile, it is a fact that variation in soil microbial populations and community structures could be distinguished by the allelopathic and non-allelopathic rice varieties tested. Our results suggest that individual components of rice root exudates, such as allelochemicals from allelopathic rice, can modify the soil microbial community.     

Structural and functional diversity of bacterial community in soil treated with the herbicide napropamide estimated by the DGGE,CLPP and r/K-strategy approaches

Napropamide is one of the most commonly used herbicide in agricultural practice and can exhibit toxic effect to soil microorganisms. Therefore, the main objective of this study was to examine the genetic and functional diversity of microbial communities in soil treated with napropamide at field rate (FR, 2.25 mg kg⁻¹ of soil) and 10 times the FR (10 × FR, 22.5 mg kg⁻¹ of soil) by the denaturing gradient gel electrophoresis (DGGE) and the community level physiological profile (CLPP) methods. In addition, the r/K-strategy approach was used to evaluate the effect of this herbicide on the community structure of the culturable soil bacteria. DGGE patterns revealed that napropamide affected the structure of microbial community; however, the richness (S) and genetic diversity (H) values indicated that the FR dosage of napropamide experienced non-significant changes. In turn, the 10 × FR dosage of herbicide caused significant changes in the S and H values of dominant soil bacteria. DGGE profiles suggest an evolution of bacteria capable of degrading napropamide among indigenous microflora. Analysis of the CLPPs indicated that the catabolic activity of microbial community expressed as AWCD (average well-color development) was temporary positively affected after napropamide application and resulted in an increase of the substrate richness (S_R) as well as functional biodiversity (H) values. Analysis of the bacterial growth strategy revealed that napropamide affected the r- or K-type bacterial classes (ecotypes). In treated-soil samples K-strategists dominated the population, as indicated by the decreased ecophysiological (EP) index. Napropamide significantly affected the physiological state of culturable bacteria and caused a reduction in the rate of colony formation as well as a prolonged time of growth rate. Obtained results indicate that application of napropamide may poses a potential risk for soil functioning.     

Characterization of cultivable heterotrophic bacterial communities in Cr-polluted and unpolluted soils using Biolog and ARDRA approaches

The bacterial communities of two soils with different chromium levels were characterized by Biolog carbon substrate utilization patterns and amplified 16S ribosomal DNA restriction analysis (ARDRA). For each bacterial community sample, cell suspensions containing 10,000 or 100 colony-forming units (CFU) were inoculated in each well of Biolog-GN microplates. The number of carbon compounds utilized by the bacterial community consisting of 100 CFU from unpolluted soil was significantly lower than that detected for the bacterial community consisting of 10,000 CFU. The size of inoculum did not substantially influence the percentage of carbon sources utilized by the Cr-polluted soil bacterial community. ARDRA approach was applied to about 100 bacterial isolates for each soil sample. A similar number of clusters for Gram-negative bacteria were found in both soils, but there were differences in percentages of isolates belonging to each group and specific genomic groups were found in each soil. Pseudomonas was the dominant taxon in both soils. Comparing the ARDRA clusters obtained from Gram-positive isolates it was evident that the culturable bacterial communities of Cr-polluted and unpolluted soils were dominated by the genus Arthrobacter and the genus Bacillus, respectively.     

Mercury affects the distribution of culturable species of Pseudomonas in soil

Pseudomonas bacteria isolated during 52 days on Gould's S1 agar from soil spiked with 0, 3.5 and 15 mg Hg(II) kg soil⁻¹ were characterised to reveal whether mercury affected them differently. Isolates from the treatments with 0 and 15 mg Hg kg⁻¹ were characterised using FT-IR characterisation and subsequent 16S rDNA partial sequencing of representative isolates. To verify the selectivity of Gould's S1 agar and the FT-IR characterisation, all 450 isolates were subjected to the following tests: Gram-determination, catalase and oxidase activity, pigment production on PDA and growth at different temperatures. Furthermore, the isolates were tested for their ability to grow on agar amended with 10 mg Hg kg⁻¹ as an indication of mercury resistance. We found that up to 80% of the isolates in soil amended with 15 mg Hg kg⁻¹ were mercury-resistant, whereas only up to 20% were resistant in the treatments with 0 and 3.5 mg Hg kg⁻¹. We found two groups of Pseudomonas, which probably represent non-described species since they did not group closely with any known species of Pseudomonas in the dendrogram. Hg-enhanced isolates were closely related to P. frederiksbergensis. Furthermore, Hg resistance was almost exclusively restricted to P. frederiksbergensis and P. migulae groups. We conclude that Hg caused a shift in the dominating species of culturable Pseudomonas.     

Influence of carbon amendments on soil denitrifier abundance in soil microcosms

It is known that carbon (C) amendments increase microbial activity in anoxic soil microcosm studies, however the effects on abundance of total and denitrifier bacterial communities is uncertain. Quantitative PCR was used to target the 16S rRNA gene for the total bacterial community, the nosZ functional gene to reflect a broad denitrifier community, and functional genes from narrow denitrifier communities represented by Pseudomonas mandelii and related species (cnorB_P) and Bosea/Bradyrhizobium/Ensifer spp. (cnorB_B). Repacked soil cores were amended with varying amounts of glucose and red clover plant tissue (0–1000 mg C kg^? 1 of soil) and incubated for 96 h. Carbon amendment significantly increased respiration as measured by cumulative CO₂ emissions. Inputs of red clover or glucose at 1000 mg C kg^? 1 of soil caused increased abundance in the total bacteria under the conditions used. There was about an approximate 2-fold increase in the abundance of bacteria bearing the nosZ gene, but only in treatments receiving 500 or 1000 mg C kg^? 1 of soil of glucose or red clover, respectively. Additions of ≥ 500 mg C kg^? 1 soil of red clover and ≥ 250 mg C kg^? 1 of glucose increased cnorB_P-gene bearing denitrifiers. Changes in abundance of the targeted communities were related to C availability in soil, as indicated by soil respiration, regardless of C source. Applications of C amendments at rates that would occur in agricultural soils not only increase microbial activity, but can also induce changes in abundance of total bacterial and denitrifier communities in studies of anoxic soil microcosms.     

Bacterial community structure and microbial activity in different soils amended with biogas residues and cattle slurry

Anaerobic digestion of organic materials generates residues of differing chemical composition compared to undigested animal manures, which may affect the soil microbial ecosystem differently when used as fertilizers. This study investigated the effects of two biogas residues (BR-A and BR-B) and cattle slurry (CS) applied at rates corresponding to 70 kg NH₄⁺-N ha⁻¹ on bacterial community structure and microbial activity in three soils of different texture (a sandy, a clay and an organic clay soil). 16S rRNA genes were targeted in PCR reactions and bacterial community profiles visualized using terminal restriction fragment length polymorphism. General microbial activity was measured as basal respiration (B-resp), substrate-induced respiration (SIR), specific growth rate (μ_SIR), metabolic quotient (qCO₂) and nitrogen mineralization capacity (NMC). Non-metric multidimensional scaling analysis visualized shifts in bacterial community structure related to microbial functions. There were significant differences in bacterial community structure after 120 days of incubation (+20 °C at 70% of WHC) between non-amended (control) and amended soils, especially in the sandy soil, where CS caused a more pronounced shift than biogas residues. Terminal-restriction fragment (TRF) 307, the predominant peak in CS-amended sandy soil, was identified as possibly Bacillus or Streptococcus. TRF 226, the dominant peak in organic soil amended with BR-B, was classified as Rhodopseudomonas. B-resp significantly increased and SIR decreased in all amendments to organic soil compared with the control, potentially indicating decreased efficiency of heterotrophic microorganisms to convert organic carbon into microbial biomass. This was also reflected in an elevated qCO₂ in the organic soil. The μ_SIR level was higher in the sandy soil amended with BR-A than with BR-B or CS, indicating a shift toward species capable of rapidly utilizing glucose. NMC was significantly elevated in the clay and organic soils amended with BR-A and BR-B and in the sandy soil amended with BR-B and CS. Thus, biogas residues and cattle slurry had different effects on the bacterial community structure and microbial activity in the three soils. However, the effects of biogas residues on microbial activities were comparable in magnitude to those of cattle slurry and the bacterial community structure was less affected. Therefore, we do not see any reason not to recommend using biogas residues as fertilizers based on the results presented.     

Vertical distribution of microbial communities under the canopy of two legume bushes in the Tehuacán Desert,Mexico

Prosopis laevigata and Parkinsonia praecox are the most abundant perennial shrubs in the Tehuacán Desert, forming ’islands of fertility’ that dominate the alluvial terraces. Both species exhibit very similar phenology, with the timing of litter foliage being the only difference between them. P. praecox litter occurs shortly after the rains, while P. laevigata maintains its leaves until the next wet season. As degradable organic matter (OM) is one of the leading factors determining soil biota composition and activity, because of the OM provided by littering, we expected that the vertical distribution of the microbial community in the vicinity of the root zone of P. praecox would be higher in comparison to P. laevigata. One soil sampling was performed; during the rainy season in August, soil samples were collected from a 0–50-cm depth at 10-cm intervals, in the vicinity of the root canopy of four individual plants of each species and the interspaces between them. Soil moisture, organic matter, and counts of bacteria and fungi under shrubs were found to decrease from the upper to deeper layers. Respiratory activity was higher in the deeper layers (p < 0.01) in all three sampling sites. Total bacterial, fungal, and heterotrophic diazotrophs were found to be significantly (p < 0.001) more numerous under shrubs than in the interspace soil. No nitrogen-fixing bacteria were isolated from interplant soils in comparison to the soil samples collected beneath the shrubs. Heterotrophic diazotrophs significantly (p < 0.01) reduced more acetylene under P. praecox (29.0 nmol/g soil) than under P. laevigata (20.1 nmol/g soil). Although the microbial numbers were unaffected by differences in plant phenology, greater nitrogenase activity under P. praecox may influence nitrogen distribution in this arid environment. Due to the fact that only one sampling was undertaken, this study elucidates the differences in the microbial community between the two shrubs, but the dynamics in the above community could not be shown.     

Effects of mercury on soil microbial communities in tropical soils of French Guyana

In gold mining regions, the risk of soil pollution by mercury is a major environmental hazard, especially in tropical areas where soil microflora plays a major part in soil functioning, major bio-geochemical cycles and carbon turn-over. The impact of mercury pollution on soil microflora should thus be carefully assessed in such environments while taking into consideration the specificities of tropical soils. The aim of this study was to compare the effects of mercury (0, 1 and 20 μg of inorganic mercury per gram of soil) on the functional diversity and genetic structure of microbial communities in a tropical soil. We investigated the effects of mercury on tropical soil microflora using soil microcosms spiked with mercury and incubated at 28 °C for 1 month. Microcosm flora, its biomass and its activity, as well as its functional and genetic structure, were followed by cultural methods, measures of respiration, ECOLOG plates, and DGGE (denaturing gel gradient electrophoresis), respectively. Fate of total and bioavailable mercury was estimated by CVAFS (cold vapor atomic fluorescence spectrometry). Results obtained for the microcosms enriched with only 1 μg g^?1 mercury were indistinguishable from controls. Conversely, in the presence of high mercury contents (20 μg g⁽¹), an immediate effect was measured on soil respiration, functional diversity (ECOLOG plates) and genetic structure (DGGE), although no significant effect was observed on plate counts or microbial biomass. In addition, whereas microbial activities (respiration and functional diversity) rapidly regained control values, a lasting effect of the high mercury concentration was observed on the genetic structure of the soil microbial community. These modifications took place during the first week of incubation when total mercury concentration was declining and bioavailable mercury was at its highest.This multiple approach study is one of the first attempts at investigating the effects of mercury on soil microbial communities in tropical soils. Our results demonstrate that in the tropical soil under study, mercury affects the soil microbial communities in a different manner than was previously reported in temperate soils. Furthermore, mercury toxicity on soil microbes may be modulated by typical tropical soil characteristics.     

Differences in soil enzyme activities,microbial community structure and short-term nitrogen mineralisation resulting from farm management history and organic matter amendments

Changes in soil microbial biomass, enzyme activities, microbial community structure and nitrogen (N) dynamics resulting from organic matter amendments were determined in soils with different management histories to gain better understanding of the effects of long- and short-term management practices on soil microbial properties and key soil processes. Two soils that had been under either long-term organic or conventional management and that varied in microbial biomass and enzyme activity levels but had similar fertility levels were amended with organic material (dried lupin residue, Lupinus angustifolius L.) at amounts equivalent to 0, 4 and 8 t dry matter lupin ha^?1. Microbial biomass C and N, arginine deaminase activity, fluorescein diacetate hydrolysis, dehydrogenase enzyme activity and gross N mineralisation were measured in intervals over an 81-day period. The community structure of eubacteria and actinomycetes was examined using PCR–DGGE of 16S rDNA fragments. Results suggested that no direct relationships existed between microbial community structure, enzyme activities and N mineralisation. Microbial biomass and activity changed as a result of lupin amendment whereas the microbial community structure was more strongly influenced by farm management history. The addition of 4 t ha^?1 of lupin was sufficient to stimulate the microbial community in both soils, resulting in microbial biomass growth and increased enzyme activities and N mineralisation regardless of past management. Amendment with 8 t lupin ha^?1 did not result in an increase proportional to the extra amount added; levels of soil microbial properties were only 1.1–1.7 times higher than in the 4 t ha^?1 treatment. Microbial community structure differed significantly between the two soils, while no changes were detected in response to lupin amendment at either level during the short-term incubation. Correlation analyses for each treatment separately, however, revealed differences that were inconsistent with results obtained for soil biological properties suggesting that differences might exist in the structure or physiological properties of a microbial component that was not assessed in this study.     

Effects of cover crops,compost, and manure amendments on soil microbial community structure in tomato production systems

Soil microbial community structure and crop yield was investigated in field tomato production systems that compared black polyethylene mulch to hairy vetch mulch and inorganic N to organic N. The following hypotheses were tested: (1) hairy vetch cover cropping increases crop yield and significantly affects soil microbial community structure when compared to the standard plastic mulch and synthetic fertilizer-based system; (2) within plastic mulch systems, organic amendments will increase crop yield and significantly affect soil microbial community structure when compared to synthetic fertilizer; (3) crop yields and microbial community structure will be similar in the hairy vetch cover cropping and the organic amended plasticulture systems. Treatments consisted of ammonium nitrate (control), hairy vetch cover crop, hairy vetch cover crop and poultry manure compost (10 Mg/ha), three levels of poultry manure compost (5, 10, and 20 Mg/ha), and two levels of poultry manure (2.5 and 5 Mg/ha). Black polyethylene mulch was used in all treatments without hairy vetch. Fatty acid analysis was used to characterize the total soil microbial community structure, while two substrate utilization assays were used to investigate the community structure of culturable bacteria and fungi. Crop yield was not significantly increased by hairy vetch cover cropping when compared to black polyethylene mulch, although microbial community structure was significantly affected by cover cropping. Under black polyethylene mulch, crop yields were significantly increased by the highest levels of compost and manure when compared to inorganic fertilizer, but there was no detectable effect on soil microbial community structure. When cover cropping was compared to organic amended plasticulture systems, crop yields were similar one year but dissimilar the next. However, hairy vetch cover cropping and organic amendments under black plastic mulch produced significantly different soil microbial community structure.     

Improving soil structure by promoting fungal abundance with organic soil amendments

Building soil structure in agroecosystems is important because it governs soil functions such as air and water movement, soil C stabilization, nutrient availability, and root system development. This study examined, under laboratory conditions, effects of organic amendments comprised of differing proportions of labile and semi-labile C on microbial community structure and macroaggregate formation in three variously textured soils where native structure was destroyed. Three amendment treatments were imposed (in order of increasing C lability): vegetable compost, dairy manure, hairy vetch (Vicia villosa Roth). Formation of water stable macroaggregates and changes in microbial community structure were evaluated over 82 days. Regardless of soil type, formation of large macroaggregates (LMA, >2000 μm diameter) was highest in soils amended with vetch, followed by manure, non-amended control, and compost. Vetch and manure had greater microbially available C and caused an increase in fungal biomarkers in all soils. Regression analysis indicated that LMA formation was most strongly related to the relative abundance of the fungal fatty acid methyl ester (FAME) 18:2ω6c (r = 0.55, p < 0.001), fungal ergosterol (r = 0.58, p < 0.001), and microbial biomass (r = 0.57, p < 0.001). Non-metric multidimensional scaling (NMS) ordination of FAME profiles revealed that vetch and manure drove shifts toward fungal-dominated soil microbial communities and greater LMA formation in these soils. This study demonstrated that, due to their greater amounts of microbially available C, vetch or manure inputs can be used to promote fungal proliferation in order to maintain or improve soil structure.     

Arable weeds,cover crops,and tillage drive soil microbial community composition in organic cropping systems

Cover crops have traditionally been used to reduce soil erosion and build soil quality, but more recently cover crops are being used as an effective tool in organic weed management. Many studies have demonstrated microbial community response to individual cover crop species, but the effects of mixed species cover crop communities have received less attention. Moreover, the relationship between arable weeds and soil microbial communities is not well understood. The objective of this study was to determine the relative influence of cover crop diversity, early-season weed communities, and tillage on soil microbial community structure in an organic cropping system through the extraction of fatty acid methyl esters (FAMEs). A field experiment was conducted between 2009 and 2011 near Mead, NE where spring-sown mixtures of zero (control), two, and eight cover crop species were included in a sunflower–soybean–corn crop rotation. A mixture of four weed species was planted in all experimental units (excluding the no-cover control), and also included as an individual treatment. Cover crops and weeds were planted in late-March, then terminated in late-May using a field disk or sweep plow undercutter, and main crops were planted within one week of termination. Three (2009) or four (2010–11) soil cores were taken to a depth of 20 cm in all experimental units at 45, 32, and 25 days following cover crop termination in 2009, 2010, and 2011, respectively. Total FAMEs pooled across 2009 and 2010 were greatest in the two species mixture–undercutter treatment combination (140.8 ± 3.9 nmol g⁻¹) followed by the eight species mixture–undercutter treatment combination (132.4 ± 3.9 nmol g⁻¹). Abundance of five (2009 and 2010) and seventeen (2011) FAME biomarkers was reduced in the weedy treatment relative to both cover-cropped treatments and the no-cover control. In 2009 and 2010, termination with the undercutter reduced abundance of most actinomycete biomarkers while termination with the field disk reduced abundance of C18:1(cis11) and iC16:0. Canonical discriminant analysis of the microbial community successfully segregated most cover crop mixture by termination method treatment combinations in 2009 and 2010. Microbial communities were most strongly influenced by the presence and type of early-spring plant communities, as weeds exerted a strong negative influence on abundance of many key microbial biomarkers, including the AMF markers C16:1(cis11) and C18:1(cis11). Weeds may alter soil microbial community structure as a means of increasing competitive success in arable soils, but this relationship requires further investigation.     

Accounting for variability in soil microbial communities of temperate upland grassland ecosystems

This study aimed to determine the factors which regulate soil microbial community organisation and function in temperate upland grassland ecosystems. Soil microbial biomass (C_mic), activity (respiration and potential carbon utilisation) and community structure (phospholipid fatty acid (PLFA) analysis, culturing and community level physiological profiles (CLPP) (Biolog^®)) were measured across a gradient of three upland grassland types; Festuca–Agrostis–Galium grassland (unimproved grassland, National Vegetation Classification (NVC) — U4a); Festuca–Agrostis–Galium grassland, Holcus–Trifolium sub-community (semi-improved grassland, NVC — U4b); Lolium–Cynosurus grassland (improved grassland, NVC — MG6) at three sites in different biogeographic areas of the UK over a period of 1 year. Variation in C_mic was mainly due to grassland type and site (accounting for 55% variance, v, in the data). C_mic was significantly (P<0.001) high in the unimproved grassland at Torridon (237.4 g C m⁻² cf. 81.2 g C m⁻² in semi- and 63.8 g C m⁻² in improved grasslands) and Sourhope (114.6 g C m⁻² cf. in 44.8 g C m⁻² semi- and 68.3 g C m⁻² in improved grasslands) and semi-improved grassland at Abergwyngregyn (76.0 g C m⁻² cf. 41.7 g C m⁻² in un- and 58.3 g C m⁻² in improved grasslands). C_mic showed little temporal variation (v=3.7%). Soil microbial activity, measured as basal respiration was also mainly affected by grassland type and site (n=32%). In contrast to C_mic, respiration was significantly (P<0.001) high in the improved grassland at Sourhope (263.4 l h⁻¹m⁻² cf. 79.6 l h⁻¹m⁻² in semi- and 203.9 l h⁻¹m⁻² unimproved grasslands) and Abergwyngregyn (198.8 l h⁻¹m⁻² cf. 173.7 l h⁻¹m⁻² in semi- and 88.2 l h⁻¹m⁻² unimproved grasslands). Microbial activity, measured as potential carbon utilisation, agreed with the respiration measurements and was significantly (P<0.001) high in the improved grassland at all three sites (A₅₉₀ 0.14 cf. 0.09 in semi- and 0.07 in unimproved grassland). However, date of sampling also had a significant (P<0.001) impact on C utilisation potential (v=24.7%) with samples from April 1997 having highest activity at all three sites. Variation in microbial community structure was due, predominantly, to grassland type (average v=23.6% for bacterial and fungal numbers and PLFA) and date of sampling (average v=39.7% for bacterial and fungal numbers and PLFA). Numbers of culturable bacteria and bacterial PLFA were significantly (P<0.001) high in the improved grassland at all three sites. Fungal populations were significantly (P<0.01) high in the unimproved grassland at Sourhope and Abergwyngregyn. The results demonstrate a shift in soil microbial community structure from one favouring fungi to one favouring bacteria as grassland improvement increased. Numbers of bacteria and fungi were also significantly (P<0.001) higher in August than any other sampling date. Canonical variate analysis (CVA) of the carbon utilisation data significantly (P<0.05) differentiated microbial communities from the three grassland types, mainly due to greater utilisation of sugars and citric acid in the improved grasslands compared to greater utilisation of carboxylic acids, phenolics and neutral amino acids in the unimproved grasslands, possibly reflecting substrate availability in these grasslands. Differences in C_mic, activity and community structure between grassland types were robust over time. In addition, broad scale measures of microbial growth and activity (C_mic and respiration) showed little temporal variation compared to measures of soil microbial community structure, which varied quantitatively with respect to environmental variables (temperature, moisture) and plant productivity, hence substrate supply.     

Biological activation of hydrous ferric oxide for reduction of hexavalent chromium in the presence of different anions

Shewanella alga BrY, a dissimilatory iron reducing bacterium (DIRB), transformed inert ferric oxides that are common in sediments, aquifer material and passivated permeable reactive iron barriers (PRBs), producing dissolved and sorbed Fe(II) capable of rapidly reducing and immobilizing Cr(VI). The effect of groundwater chemistry on the formation and reactivity of such microbial-produced, abiotic reductants was investigated. Batch reactors with high carbonate concentration (10 mM) were the most reactive, removing 66.0% ± 2.8 of Cr (VI) (76 mg/l) from liquid phase within 5 min. Treatments with high concentrations of sulfate (5.2 mM), chloride (10 mM), phosphate (1 mM) or silica (0.75 mM) were less reactive (about 40% removal). Loss of reactivity was observed possibly due to oxidation of Fe(II) (sorbed and dissolved) by Cr(VI). Normalization of Cr(VI) removal to the mass of biogenic solid present showed the following molar Cr/Fe ratios in solid phase: 0.185 ± 0.041 (carbonate), 0.146 ± 0.013 (sulfate), 0.092 ± 0.010 (silica), 0.075 ± 0.012 (phosphate) and 0.062 ± 0.012 (chloride). Overall, these results show that bacterial transformation of inert ferric oxides can contribute to the (abiotic) natural attenuation of Cr(VI) in and around PRBs, and that groundwater chemistry is an important determinant of biogenic solids reactivity.     

The effects of fresh and stabilized pruning wastes on the biomass,structure and activity of the soil microbial community in a semiarid climate

The incorporation of organic amendments from pruning waste into soil may help to mitigate soil degradation and to improve soil fertility in semiarid ecosystems. However, the effects of pruning wastes on the biomass, structure and activity of the soil microbial community are not fully known. In this study, we evaluate the response of the microbial community of a semiarid soil to fresh and composted vegetal wastes that were added as organic amendments at different doses (150 and 300 t ha⁻¹) five years ago. The effects on the soil microbial community were evaluated through a suite of different chemical, microbiological and biochemical indicators, including enzyme activities, community-level physiological profiles (CLPPs) and phospholipid fatty acid analysis (PLFA). Our results evidenced a long-term legacy of the added materials in terms of soil microbial biomass and enzyme activity. For instance, cellulase activity reached 633 μg and 283 μg glucose g⁻¹ h⁻¹ in the soils amended with fresh and composted waste, respectively. Similarly, bacterial biomass reached 116 nmol g⁻¹ in the soil treated with a high dose of fresh waste, while it reached just 66 nmol g⁻¹ in the soil amended with a high dose of composted waste. Organic amendments produced a long-term increase in microbiological activity and a change in the structure of the microbial community, which was largely dependent on the stabilization level of the pruning waste but not on the applied dose. Ultimately, the addition of fresh pruning waste was more effective than the application of composted waste for improving the microbiological soil quality in semiarid soils.     

Tag-encoded pyrosequencing analysis of bacterial diversity in a single soil type as affected by management and land use

Impacts of management and land use on soil bacterial diversity have not been well documented. Here we present the application of the bacterial tag encoded FLX amplicon pyrosequencing (bTEFAP) diversity method, which will promote studies in soil microbiomes. Using this modified FLX pyrosequencing approach we evaluated bacterial diversity of a soil (Pullman soil; fine, mixed, thermic Torrertic Paleustolls) with 38% clay and 34% sand (0–5 cm) under four systems. Two non-disturbed grass systems were evaluated including a pasture monoculture (Bothriochloa bladhii (Retz) S.T. Blake) [P] and a diverse mixture of grasses in the Conservation Reserve Program (CRP). Two agricultural systems were evaluated including a cotton (Gossypium hirsutum L.) -winter wheat (Triticum aestivum L.)-corn (Zea mays L.) rotation [Ct–W–Cr] and the typical practice of the region, which is continuous monoculture cotton (Ct–Ct). Differences due to land use and management were observed in soil microbial biomass C (CRP > P = Ct–W–Cr > Ct–Ct). Using three estimators of diversity, the maximum number of unique sequences operational taxonomic units (OTU; roughly corresponding to the species level) never exceeded 4500 in these soils at the 3% dissimilarity level. The following trend was found using the most common estimators of bacterial diversity: Ct–W–Cr > P = CRP > Ct–Ct. Predominant phyla in this soil were Actinobacteria, Bacteriodetes and Fermicutes. Bacteriodetes were more predominant in soil under agricultural systems (Ct–W–Cr and Ct–Ct) compared to the same soil under non-disturbed grass systems (P and CRP). The opposite trend was found for the Actinobacteria, which were more predominant under non-disturbed grass systems (P and CRP). Higher G? bacteria and lower G+ bacteria were found under Ct–W–Cr rotation and highest abundance of actinomycetes under CRP. The bTEFAP technique proved to be a powerful method to characterize the bacterial diversity of the soil studied under different management and land use in terms not only on the presence or absence, but also in terms of distribution.     

Impact of bovine urine deposition on soil microbial activity,biomass, and community structure

Nitrogen (N) from urine excreted by grazing animals can be transformed into N compounds that have detrimental effects on the environment. These include nitrate, which can cause eutrophication of waterways, and nitrous oxide, which is a greenhouse gas. Soil microbes mediate all of these N transformations, but the impact of urine on microbes and how initial soil conditions and urine chemical composition alter their responses to urine are not well understood. This study aimed to determine how soil inorganic N pools, nitrous oxide fluxes, soil microbial activity, biomass, and the community structure of bacteria containing amoA (nitrifiers), nirK, and nirS (denitrifiers) genes responded to the addition of urine over time. Bovine urine containing either a high (15.0 g K⁺ l^?1) or low salt content (10.4 g K⁺ l^?1) was added to soil cores at either low or high moisture content (hereafter termed dry and wet soil respectively; 35% or 70% water-filled pore space after the addition of urine). Changes in soil conditions, inorganic N pools, nitrous oxide fluxes, and the soil microbial community were then measured 1, 3, 8, 15, 29 and 44 days after urine addition. Urine addition increased soil ammonium concentrations by up to 2 mg g d.w.^?1, soil pH by up to 2.7 units, and electrical conductivity (EC) by 1.0 and 1.6 dS m^?1 in the low and high salt urine treatments respectively. In response, nitrate accumulation and nitrous oxide fluxes were lower in dry compared to wet urine-amended soils and slightly lower in high compared to low salt urine-amended soils. Nitrite concentrations were elevated (>3 μg g d.w.^?1) for at least 15 days after urine addition in wet urine-amended soils, but were only this high in the dry urine-amended soils for 1 day after the addition of urine. Microbial biomass was reduced by up to half in the wet urine-amended soils, but was largely unaffected in the dry urine-amended soils. Urine addition affected the community structure of ammonia-oxidising and nitrite-reducing bacteria; this response was also stronger and more persistent in wet than in dry urine-amended soils. Overall, the changes in soil conditions caused by the addition of urine interacted to influence microbial responses, indicating that the effect of urine on soil microbes is likely to be context-dependent.     

Effects of long-term sewage irrigation on agricultural soil microbial structural and functional characterizations in Shandong,China

Soil samples taken from a sewage irrigation area, a partial sewage irrigation area and a ground water irrigation area (control area) were studied with the methods of Biolog and FAME. It was found that the microbial utilization of carbon sources in sewage irrigation areas was much higher than that of control area (P < 0.05). With the increasing of the amount of sewage irrigation, microbial functional diversity slightly increased by the Biolog analysis; however, the amount of epiphyte decreased by the FAME analysis. The results also showed that the Cr, Zn contents were positively correlated with the values of AWCD and the microbial diversity, while Hg content showed negative correlation with the microbial parameters (AWCD of 72 h and Shannon index). Our studies suggested that sewage irrigation resulted in an obvious increase of heavy metals content in soil (P < 0.05), although the maximum heavy metals concentrations were much lower than the current standard of China. Other soil basic characteristics such as cation exchange capacity (CEC), total nitrogen (N_t) and organic matter in sewage irrigation areas obviously increased (P < 0.05). Therefore, it is demonstrated that long-term sewage irrigation had influenced soil microorganisms and soil quality in the studied soils. As a result, it is important to monitor the changes in agricultural soils. Furthermore, our results also confirmed that the methods of Biolog and FAME are effective tools for the assessment of soil microbial structure/function and soil health.     

Soil microbial community responses to sulfadiazine-contaminated manure in different soil microhabitats

Veterinary antibiotics such as sulfadiazine (SDZ) are applied with manure to agricultural soil. Antimicrobial effects of SDZ on soil microbial community structures and functions were reported for homogenized bulk soils. In contrast, field soil is structured. The resulting microhabitats are often hot spots that account for most of the microbial activity and contain strains of different antibiotic sensitivity or resilience. We therefore hypothesize that effects of SDZ are different in diverse soil microhabitats. We combined the results of laboratory and field experiments that evaluated the fate of SDZ and the response of the microbial community in rhizosphere, earthworm burrow, and soil macroaggregate microhabitats. Microbial communities were characterized by phenotypic phospholipid fatty acid (PLFA) and genotypic 16S rRNA gene patterns (DGGE) and other methods. Data was evaluated by principle component analyses followed by two-way ANOVA with post-hoc tests. Extractable SDZ concentrations in rhizosphere soil were not clearly different and varied by a factor 0.7–1.2 from those in bulk soil. In contrast to bulk soil, the extractable SDZ content was two-fold larger in earthworm burrows, which are characterized by a more hydrophobic organic matter along the burrow surface. Also, extractable SDZ was larger by up to factor 2.6 in the macroaggregate surface soil. The rhizosphere effect clearly increased the microbial biomass. Nonetheless, in the 10 mg SDZ kg⁻¹ treatment, the biomass deceased by about 20% to the level of uncontaminated bulk soil. SDZ contamination lowered the total PLFA concentrations by 14% in the rhizosphere and 3% in bulk soil of the field experiment. Structural shifts represented by Pseudomonas DGGE data were larger in SDZ-contaminated earthworm burrows compared to bulk soils. In the laboratory experiment, a functional shift was indicated by a four-fold reduced acid phosphatase activity in SDZ-contaminated burrows compared to bulk soil. Structural and functional shifts after SDZ contamination were larger by a factor of 2.5 in the soil macroaggregate surface versus interior, but this relation reversed over the long-term under field conditions. Overall, the combined effects of soil microhabitat, microbial community composition, and exposure to SDZ influenced the microbial susceptibility towards antibiotics under laboratory and field conditions.