Sequencing Zerovalent Iron Treatment with Carbon Amendments to Remediate Agrichemical-Contaminated Soil

Agrichemical spills and discharges to soil can cause point-source contamination of surface and ground waters. When high contaminant concentrations inhibit natural attenuation in soils, chemical treatments can be used to promote degradation and allow application of treated soils to agricultural lands. This approach was used to remediate soil containing >650 mg atrazine, >170 mg metolachlor and >18,000 mg nitrate kg^?1. Results indicated a decrease in metolachlor concentration to <1 mg kg^?1 within 95 days of chemical treatment with zerovalent iron (Fe⁰, 5% w/w) and aluminum sulfate (Al₂(SO₄)₃, 2% w/w) but after one year >150 mg atrazine and >7000 mg nitrate kg^?1 remained. Laboratory experiments confirmed that subsequent additions of sucrose (table sugar) to the chemically pretreated soil promoted further reductions in atrazine and nitrate concentrations. Field-scale results showed that adding 5% (w/w) sucrose to windrowed and pretreated soil significantly reduced atrazine (<38 mg kg^?1) and nitrate (<2,100 mg kg^?1) concentrations and allowed for land application of the treated soil. These results provide evidence that zerovalent iron in combination with Al₂(SO₄)₃ and sucrose can be used for on-site, field-scale treatment of pesticide- and nitrate-contaminated soil.     

Modifications to atrazine degradation pathways in a loamy soil after addition of organic amendments

The effects of two organic amendments, a municipal solid waste compost and a composted straw, on [U-ring- ]atrazine degradation pathways in a loamy soil (Grignon, Yvelines, France) were studied during laboratory incubations under controlled conditions. Three month conditionings were conducted under sterile or non-sterile conditions, with soil plus atrazine, organic amendment plus atrazine, or soil plus organic amendment. Then either an organic amendment or soil or atrazine was added, respectively, to these three treatments and incubated for an additional 3 months under non-sterile conditions. Both organic amendments modified the behaviour of atrazine in soil but via different processes. The addition of municipal compost increased atrazine sorption and decreased its availability for degradation by soil microorganisms. The effect of the composted straw was mainly related to its high enzymatic activity, which appeared to be responsible for the production of large amounts of hydroxyatrazine. This hydroxylation favoured the opening of the triazine ring and its subsequent mineralization in the soil. At the end of the incubations, less atrazine was mineralized in the presence of the two types of organic amendment, which both increased the formation of non-extractable residues of atrazine. The addition of municipal compost preserved larger amounts of extractable atrazine, while the addition of composted straw enhanced hydroxyatrazine production. In all cases, the greatest effects were found when atrazine was directly in contact with the organic amendment during conditioning.     

Molecular and physiological bacterial diversity of a semi-arid soil contaminated with different levels of formulated atrazine

A semi-arid soil treated with different concentrations of formulated atrazine in a laboratory experiment was studied over 45 days, by different biological and molecular parameters (bacterial enumeration (cfu), community level physiological profiles (CLPPs) measured by Biolog^® and denaturing gradient gel electrophoresis (DGGE)), to study the bacterial community diversity.Formulated atrazine was almost totally degraded at different concentrations after this incubation time. The number of colony forming units (cfu) for soils with 100 and 1000 mg kg⁻¹ atrazine was significantly (p ≤ 0.05) higher than for the control, 1 and 10 mg kg⁻¹ treatments. DGGE banding patterns showed that regardless of time elapsed, concentrations of 10, 100 and 1000 mg kg⁻¹ atrazine in soil, affected the bacterial community compared to control and 1 mg kg⁻¹.The Shannon diversity index (H′) based on CLPP data showed a significant (p ≤ 0.05) decrease at atrazine concentrations of 100 and 1000 mg kg⁻¹. The Shannon diversity indices for different guilds of source carbon and the parameters K and r (based on the kinetics of colour formation rather than on the degree of colour development) were related to guilds of carbon substrates and atrazine concentration at a sampling time. The parameter K was very sensitive to atrazine effects on microbial communities.These biological and molecular parameters can be used to monitor changes in soils treated with atrazine at different concentrations, even when the pesticide is degraded.     

Influence of nitrogen on atrazine and 2, 4 dichlorophenoxyacetic acid mineralization in blackwater and redwater forested wetland soils

 Microcosms were used to determine the influence of N additions on active bacterial and fungal biomass, atrazine and dichlorophenoxyacetic acid (2,4-D) mineralization at 5, 10 and 15 weeks in soils from blackwater and redwater wetland forest ecosystems in the northern Florida Panhandle. Active bacterial and fungal biomass was determined by staining techniques combined with direct microscopy. Atrazine and 2,4-D mineralization were measured radiometrically. Treatments were: soil type, (blackwater or redwater forested wetland soils) and N additions (soils amended with the equivalent of 0, 200 or 400 kg N ha^–1 as NH₄NO₃). Redwater soils contained higher concentrations of C, total N, P, K, Ca, Mn, Fe, B and Zn than blackwater soils. After N addition and 15 weeks of incubation, active bacterial biomass in redwater soils was lower when N was added. Active bacterial biomass in blackwater soils was lower when 400 kg N ha^–1, but not when 200 kg N ha^–1, was added. Active fungal biomass in blackwater soils was higher when 400 kg N ha^–1, but not when 200 kg N ha^–1, was added. Active fungal biomass in redwater soils was lower when 200 kg N ha^–1, but not when 400 kg N ha^–1, was added. After 15 weeks of incubation 2,4-D degradation was higher in redwater wetland soils than in blackwater soils. After 10 and 15 weeks of incubation the addition of 200 or 400 kg N ha^–1 decreased both atrazine and 2,4-D degradation in redwater soils. The addition of 400 kg N ha^–1 decreased 2,4-D degradation but not atrazine degradation in blackwater soils after 10 and 15 weeks of incubation. High concentrations of N in surface runoff and groundwater resulting from agricultural operations may have resulted in the accumulation of N in many wetland soils. Large amounts of N accumulating in wetlands may decrease mineralization of toxic agricultural pesticides. Received: 26 June 1998     

Atrazine degradation in boreal nonagricultural subsoil and tropical agricultural soil

Purpose

The purpose of this study was to determine the natural atrazine degradation activity and the genetic potential in a soil profile spanning down to the groundwater zone, collected in Finland at a site where past use of atrazine has contaminated the groundwater, and in Indian agricultural topsoils having different histories of atrazine use.

Materials and methods

Atrazine degradation potential was assessed by quantifying the atrazine degradation genes atzA, trzN, and atzB by quantitative PCR reaction. Atrazine mineralization was studied by radiorespirometry in order to find out if these genes were expressed.

Results and discussion

Indian soils contained a large number up to 10⁴–10⁵ copies (g^?1 dry weight (dw) soil) of atrazine degradation genes after the first treatment with atrazine. These genes were also expressed, as up to 55 % of atrazine mineralized. Some unspecific binding of primers required thorough investigation and confirmation by sequencing of the qPCR products in the agricultural soil samples. The degradation capability of the nonagricultural boreal soil profile was much lower: atrazine degradation genes were present at detection limit (10² copies g^?1 soil), but mineralization studies indicated that these genes were not transcribed, since no or very little atrazine mineralization was observed.

Conclusions

Our results indicate that when atrazine was applied in agricultural practice, the soil atrazine degradation capacity was high. The organisms responsible for the degradation were effectively degrading atrazine already 3 months after the first treatment with atrazine. However, in boreal soil, decades after atrazine use had been discontinued, residual atrazine was not degraded even though a small number of degradation genes could still be detected in soil. There is a need for more specific primers for qPCR in tropical soils.     

How increasing availabilities of carbon and nitrogen affect atrazine behaviour in soils

 The effect of increasing amounts of glucose and mineral N on the behaviour of atrazine was studied in two soils. One had been exposed to atrazine under field conditions (adapted soil), the other had not (non-adapted soil), resulting, respectively, in an accelerated degradation of atrazine in the adapted soil and in a slow degradation of the herbicide in the non-adapted soil. The dissipation of ¹⁴C-atrazine via degradation and formation of non-extractable "bound" residues was followed during laboratory incubations in soils supplemented or not with increasing amounts of glucose and mineral N. In both soils, glucose added at rates of up to 16 g C kg^–1 soil did not modify atrazine mineralization but increased the formation of bound residues; this was probably due to the retention of atrazine by the growing microbial biomass. Atrazine dealkylation was enhanced when a large amount of glucose was added. In both soils, the addition of the largest dose of mineral N (2.5 g N kg^–1 soil) decreased atrazine mineralization. The simultaneous addition of glucose and mineral N enhanced their effects. When the largest doses of mineral N and glucose were added, atrazine mineralization stopped in both soils, and the proportion of bound residues increased. Glucose and mineral N additions influenced atrazine mineralization to a greater extent in the adapted soil than in the non-adapted one, as revealed by ANOVA, although glucose addition had a greater effect than N. The competition for space and nutrients between atrazine-degrading microorganisms and the total heterotrophic microflora probably contributed to the decrease in atrazine mineralization. Received: 9 June 1998     

Mineralization of the herbicide MCPA in urban soils is linked to presence and growth of class III tfdA genes

Mineralization and sorption of ¹⁴C-ring labeled herbicide 2-methyl-4-chlorophenoxyacetic acid (MCPA) were quantified along with the tfdA gene abundance in 7 different soils. The soils tested were five gravel soils from urban locations, one soil from the embankment of a railway track, and finally an agricultural soil as a control. The mineralization experiments were performed with a concentration of MCPA of 5 mg/kg and incubated at 10 °C for a period of 60 days.With K_d values ranging from 0.04 to 0.41 l kg⁻¹ the sorption experiments revealed that binding of MCPA to the six gravel soils was lower compared to the control soil which had a K_d value of 0.91 l kg⁻¹. The potential for MCPA mineralization varied from less than 5 to over 55% mineralized in 60 days. The most rapid MCPA mineralization was observed in the soil from the Danish railway tracks with 55% mineralized after only 18 days. The mineralization data was fitted to degradation kinetic models, which indicated that growth occurred as a response to MCPA degradation in most of the soils.Soil DNA was extracted and tfdA genes responsible for the first step in MCPA degradation were quantified by real-time PCR (qPCR) at appropriate time points throughout the mineralization experiments. Indicating growth of specific MCPA degraders, the abundance of class III tfdA genes showed an increase during MCPA mineralization in those soils able to mineralize MCPA.These findings emphasize the importance of the presence of microorganisms that are able to readily degrade MCPA, to avoid groundwater leaching following use on urban gravel areas that possess low binding ability of the compound.     

Persistence of Atrazine and Alachlor in Ground Water Aquifers and Soil

Degradation of atrazine and alachlor in saturated aquifer materials and soil was studied in the laboratory. A static aquifer was represented by a set of stagnant flasks and a well-mixed aquifer was simulated by recirculating columns. Water was tested at selected time intervals over six months and analyzed for herbicides and metabolites. Under all conditions, atrazine was more persistent than alachlor. Increased temperature had little effect on atrazine dissipation but did increase alachlor degradation rates, especially in the sterilized treatments. The addition of carbon and nitrogen prolonged the initial period before the onset of degradation in some of the columns. Enhanced mass transfer of the herbicides, nutrients, and oxygen in the recirculating columns dramatically increased dissipation of atrazine and alachlor. The degradation rates of atrazine and alachlor were 2 to 5 times faster in the recirculating columns than in the stagnant flasks. Atrazine was more persistent in the aquifer materials than in the soils, while alachlor dissipation was similar in the soils and recirculating aquifer columns, but was slower in the stagnant flasks. The prolonged persistence of atrazine under static, aquifer conditions (t _1/2 = 206 to 710 days) indicates that natural mechanisms are not sufficient to alleviate the risk of atrazine buildup over time; however, in a well mixed aquifer, atrazine degradation rates should be higher (t _1/2 = 66 to 106 days) and the threat of atrazine accumulation is diminished. Alachlor persistence at low concentrations (< 10 μg L^?1) in aquifers should not pose a long-term threat to ground water supplies.     

THE ADSORPTION OF COPPER BY SOIL MATERIALS AT LOW EQUILIBRIUM SOLUTION CONCENTRATIONS

The adsorption of copper by individual soil components (organic matter fractions, oxides and clay minerals) was examined at equilibrium solution concentrations of copper within the range found in natural soils, the distribution of copper between solution and solid phases being measured by means of labelling with radioactive ⁶⁴Cu. At these low solution concentrations it was found that the copper adsorption isotherms were essentially linear. The oxides and organic materials adsorbed the greatest amounts of copper. The concentration of copper in natural soil solutions will be controlled by these materials to a far greater extent than by the clay minerals, the influence of which may be negligible in some soils. Solution concentrations of copper are relatively unaffected by both the background concentration of major cations and by changes in pH within the ionic strength and pH range found in normal agricultural soils. Copper adsorption studies with humic and fulvic acids showed that total solution copper concentrations could be greatly enhanced above the equilibrium levels for ionic copper by the presence of soluble organic complexes. The importance of taking into account the presence of such copper complexes in soil copper studies is emphasised.     

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.     

Biochemical properties and microbial community structure of five different soils after atrazine addition

Atrazine is one of the most used herbicides worldwide; however, consequences of its long-term agricultural use are still unknown. A laboratory study was performed to examine changes in microbial properties following ethylamino-¹⁵N-atrazine addition, at recommended agronomic dose, to five acidic soils from Galicia (NW Spain) showing different physico-chemical characteristics, as well as atrazine application history. Net N mineralization was observed in all soils, with nitrate being the predominant substance formed. The highest values were detected in soils with low atrazine application history. From 2% to 23% of the atrazine-¹⁵N was found in the soil inorganic-N pool, the highest values being detected after 9 weeks in soils with longer atrazine application history and lower indigenous soil N mineralization. The application of atrazine slightly reduced the amount of soil N mineralized and microbial biomass at short term. Soluble carbohydrates and β-glucosidase and urease activity decreased with incubation time, but were not significantly affected by the single application of atrazine. Microbial community structure changed as consequence of both soil type and incubation time, but no changes in the phospholipid fatty acid (PLFA) pattern were detected due to recent atrazine addition at normal doses. The saturated 17- to 20-carbon fatty acids had higher relative abundance in soils with a longer atrazine history and fungal biomass, as indicated by the PLFA 18:2ω6,9, decreased with the incubation time. The results suggested that the PLFA pattern and soil N dynamics can detect the long-term impact of repeated atrazine application to agricultural soils.     

Relationships between carbon dioxide emission and soil properties in salt-affected landscapes

Net carbon dioxide (CO₂) emission from soils is controlled by the input rate of organic material and the rate of decomposition which in turn are affected by temperature, moisture and soil factors. While the relationships between CO₂ emission and soil factors are well-studied in non-salt-affected soils, little is known about soil properties controlling CO₂ emission from salt-affected soils. To close this knowledge gap, non-salt-affected and salt-affected soils (0-0.30 m) were collected from two agricultural regions: in India (irrigation induced salinity) and in Australia (salinity associated with ground water or non-ground water associated salinity). A subset (50 Indian and 70 Australian soils) covering the range of electrical conductivity (EC) and sodium adsorption ratio (SAR) in each region was used in a laboratory incubation experiment. The soils were left unamended or amended with mature wheat residues (2% w/w) and CO₂ release was measured over 120 days at constant temperature and soil water content. Residues were added to overcome carbon limitation for soil respiration. For the unamended soils, separation in multidimensional scaling plots was a function of differences in soil texture (clay, sand), SOC pools (particulate organic carbon (POC) and humus-C) and also EC. Cumulative CO₂-C emission from unamended and amended soils was related to soil properties by stepwise regression models. Cumulative CO₂-C emission was negatively correlated with EC in saline soils (R² = 0.50, p < 0.05) from both regions. In the unamended non-salt-affected soils, cumulative CO₂-C emission was significantly positively related to the content of POC for the Indian soils and negatively related to clay content for the Australian soils. In the wheat residue amended soils, cumulative CO₂-C emission had positive relationship with POC and humus-C but a negative correlation with EC for both Indian and Australian soils. SAR was negatively related (β = −0.66, p < 0.05) with cumulative CO₂-C emission only for the unamended saline-sodic soils of Australia. Cumulative CO₂-C emission was significantly negatively correlated with bulk density in amended soils from both regions. The study showed that in salt-affected soils, EC was the main factor influencing for soil respiration but the content of POC, humus-C and clay were also influential with the magnitude of influence depending on whether the soils were salt affected or not.     

阿特拉津在土壤, 矿物质及堆肥中的吸附, 运输和转化

Atrazine is a widely used herbicide for controlling weeds on both agricultural and nonagricultural land,which is equally detected in water supplies beyond safe concentrations.Although the presence of atrazine metabolites is an indication of herbicide degradation,some of them still exhibit toxicity,greater water solubility and weaker interaction with soil components than atrazine.Hence,studies with atrazine in the environment are of interest because of its potential to contaminate drinking water sources.Data on atrazine availability for transport,plant uptake,and microbial degradation and mineralization are therefore required to perform more comprehensive and realistic environmental risk assessments of its environmental fate.This review presents an account of the sorption-desorption phenomenon of atrazine on soil and other sorbents by revisiting the several mechanisms of atrazine-sorbent binding reported in the literature.The retention and transport of atrazine in soils;the influence of organic matter on atrazine sorption;the interactions of atrazine with humic substances,atrazine uptake by plants,atrazine bioccumulation and microbial degradation;atrazine transformation in composting environments;and finally atrazine removal by biosorption are discussed.     

Differential copper impact on density,diversity and resistance of adapted culturable bacterial populations according to soil organic status

The effect of copper on the abundance, diversity and resistance of viable heterotrophic and copper resistant bacterial populations (Cu^R) was evaluated in soils differing only by their amount and type of organic matter. These soils have been obtained using a vineyard soil that had been subjected to three different organic matter managements (Not Amended (NA) or amended with Straw (S) or Conifer Compost (CC)) in a long term field experiment. Soil microcosms were artificially contaminated with copper (250 mg Cu kg^?1 of soil) and incubated for 35 days. Throughout the incubation, a differential copper impact on viable heterotrophic and Cu^R bacterial enumeration was demonstrated according to the soil organic status with a magnitude which followed the order: NA > CC ≥ S. Diversity of Cu^R bacteria revealed no significant difference between the uncontaminated soils, as determined by 16S rRNA sequencing. However, copper spiking induced an enrichment of particular populations depending on soil, with Methylobacterium, Ralstonia and Staphylococcus like species becoming dominant in NA, S and CC soils, respectively. Evaluation of the copA gene distribution and diversity, through PCR detection and sequencing, revealed that few Cu^R bacteria (from 7 to 13%) possessed this genetic determinant before the addition of metal. Copper contamination induced an increase in the dissemination of homologous copA genes only in Ralstonia like species indigenous to S soils. From a functional point of view, copper minimum inhibitory concentration for each Cu^R strain was measured. It did not highlight variable copper resistance efficiency between strains belonging to different taxonomic groups, harboring or not the copA gene, and originating from different soils contaminated or not by copper.     

Glufosinate and ammonium sulfate inhibit atrazine degradation in adapted soils

The co-application of glufosinate with nitrogen fertilizers may alter atrazine cometabolism, thereby extending the herbicide’s residual weed control in adapted soils. The objective of this study was to assess the effects of glufosinate, ammonium sulfate, and the combination of glufosinate and ammonium sulfate on atrazine mineralization in a Dundee silt loam exhibiting enhanced atrazine degradation. Application of glufosinate at rates of 10 to 40 mg kg⁻¹ soil extended the lag phase 1 to 2 days and reduced the maximum degradation rate by 15% to 30%. However, cumulative atrazine mineralization averaged 85% 21 days after treatment and was independent of treatment. Maximum daily rates of atrazine mineralization were reduced from 41% to 55% by application of 1 to 8 g kg⁻¹ of ammonium sulfate. Similarly, cumulative atrazine mineralization was inversely correlated with ammonium sulfate rates ranging from 1.0 to 8 g kg⁻¹ soil. Under the conditions of this laboratory study, atrazine degradation was relatively insensitive to exogenous mineral nitrogen, in that 8 g (NH₄)₂SO₄ per kilogram soil repressed but did not completely inhibit atrazine mineralization. Moreover, an additive effect on reducing atrazine mineralization was observed when glufosinate was co-applied with ammonium sulfate. In addition, ammonium fertilization alters the partitioning of ¹⁴C-atrazine metabolite accumulation and nonextractable residues, indicating that ammonium represses cleavage of the triazine ring. Consequently, results indicate that the co-application of glufosinate with N may increase atrazine persistence under field conditions thereby extending atrazine residual weed control in adapted soils.     

N-dealkylation of atrazine by hydrogenotrophic fluorescent Pseudomonads

Two strains of Pseudomonas fluorescens, LMG 10141 and 10140, isolated from soil, cometabolised atrazine in water under hydrogenotrophic conditions. The degradation products were deethylatrazine and deisopropylatrazine while hydroxyatrazine was not detected in cultures of LMG 10141. Labelled experiments indicated that neither the alkyl chains nor the ring structure were completely mineralised.     

Soil microbial activity measured by microcalorimetry in response to long-term fertilization regimes and available phosphorous on heat evolution

Microcalorimetric technique was used to investigate the effects of balanced versus nutrient-deficiency fertilization on soil microbial activity in a long-term (19-year) fertilizer experiment. The number of microorganisms in soils was measured by viable cell count, and the power-time curves were recorded for soil samples supplemented with glucose and ammonium sulphate, also with or without sodium dihydrogen phosphate for P-deficiency fertilization. Both the bacterial and the fungal populations were significantly higher (P < 0.05) under balanced fertilization than under nutrient-deficiency fertilization. The microbial activity presented by heat dissipation per cell unit indicated that microorganisms under balanced fertilizer treatments had more efficient metabolism, while decreased microbial activity under nutrient-deficiency treatments was firstly due to soil available P, followed by N and K (P < 0.05). In addition, microbial growth in soils under P-deficiency fertilization was stimulated by adding available P, while the lower growth rate, less peak heat, and longer peak time all indicated the low activity of soil microorganisms. We emphasize the importance of balanced fertilization, as well as the role of available P, in maintaining and promoting soil microbial activity.     

Laboratory Study of Leaching Properties of Mediterranean Forest Species Ashes

For bioremediation of copper-contaminated soils, it is essential to understand copper adsorption and chemical forms in soils related to microbes. In this study, a Penicillium strain, which can tolerate high copper concentrations up to 150 mmol l^?1 Cu²⁺, was isolated from a copper mining area. The objective was to study effects of this fungus on copper adsorptions in solutions and chemical forms in soils. Results from lab experiments showed the maximum biosorptions occurred at 360 min with 6.15 and 15.08 mg g^?1 biomass from the media with Cu²⁺ of 50 and 500 mg l^?1, respectively. The copper was quickly adsorbed by the fungus within the contact time of the first 60 min. To characterize the adsorption process of copper, four types of kinetics models were used to fit the copper adsorption data vs. time. Among the kinetics models, the two-constant equation gave the best results, as indicated by the high coefficients of determination (R ²?=?0.89) and high significance (p?<?0.01). The addition of the fungal strain to autoclaved soil facilitated increases in concentrations of acid-soluble copper, copper bound to oxides, and of copper bound to organic matter (p?<?0.05). However, the inoculation of Penicillium sp. A1 led to a decrease of water-soluble copper in the soil. The results suggested that Penicillium sp. A1 has the potential for bioremediation of copper-contaminated soils.     

Assessing impacts of copper on soil enzyme activities in regard to their natural spatiotemporal variation under long-term different land uses

Enzymes are known to be sensitive indicators of soil quality. Nevertheless, the natural variability of their activities needs to be considered for a relevant interpretation of activity levels, especially in contaminated soils. We first monitored the variability of enzymatic activities (acid and alkaline phosphatases, β-glucosidase, N-acetyl-β-glucosaminidase, urease and dehydrogenase) in two agricultural sites in north-western France during a seasonal April–October cycle. For both sites, two types of long-term different agricultural managements, grassland and intensive cropping were considered. Our results revealed great variability of enzymatic activities in space and over time, yet more pronounced for grassland than for cropped soils. Then, we assessed the impact of copper on enzymes activity on Terrestrial Model Ecosystems (TMEs) filled with undisturbed soil, and incubated for 70 days in open-field conditions. Copper was added at two concentration levels corresponding to a regulated annual agronomic input (2 mg kg⁻¹) or to a high soil contamination (200 mg kg⁻¹). In comparison to effects of natural spatiotemporal variability of soil conditions, copper addition did not show significant impacts on enzymatic activities. Finally, our results confirmed that for assessing effective impacts of contaminants in soils under real field conditions, natural spatiotemporal soil variability must be considered.     

Microbial community PLFA and PHB responses to ecosystem restoration in tallgrass prairie soils

Native North American prairie grasslands are renowned for the richness of their soils, having excellent soil structure and very high organic content and microbial biomass. In this study, surface soils from three prairie restorations of varying ages and plant community compositions were compared with a nearby undisturbed native prairie remnant and a cropped agricultural field in terms of soil physical, chemical and microbial properties. Soil moisture, organic matter, total carbon, total nitrogen, total sulfur, C:N, water-holding capacity and microbial biomass (total PLFA) were significantly greater (p<0.05) in the virgin prairie remnant as well as the two long-term (21 and 24 year) prairie restorations, compared with the agricultural field and the restoration that was begun more recently (7 years prior to sampling). Soil bulk density was significantly greater (p<0.05) in the agricultural and recently restored sites. In most cases, the soil quality indicators and microbial community structures in the restoration sites were intermediate between those of the virgin prairie and the agricultural sites. Levels of poly-β-hydroxybutyrate (PHB) and PLFA indicators of nutritional stress were significantly greater (p<0.05) in the agricultural and recent restoration sites than in the long-term restorations or the native prairie. Samples could be assigned to the correct site by discriminant analysis of the PLFA data, with the exception that the two long-term restoration sites overlapped. Redundancy analysis showed that prairie age (p<0.005) was the most important environmental factor in determining the PLFA microbial community composition, with C:N (p<0.015) also being significant. These findings demonstrate that prairie restorations can lead to improved quality of surface soils. We predict that the conversion of farmland into prairie will shift the soil quality, microbial community biomass and microbial community composition in the direction of native prairies, but with the restoration methods tested it may take many decades to approach the levels found in a virgin prairie throughout the soil profile.