Feasibility of a counter-current extraction procedure for the removal of heavy metals from contaminated soils

Effective remediation and sanitation technologies for soils contaminated with heavy metals are limited. We investigated the feasibility of a counter-current metal extraction procedure for the removal of selected heavy metals (Cd, Cu, Ph, and Zn) from two contaminated soils. The process involved a decarbonation (removal of carbonates), acid solubilisation, washing, and liming step. Results from batch equilibration experiments simulating the counter-current process showed more than 85% of the Cd present to be removed. Removal efficiencies for Cu and Pb were limited to approximately 15%, this mainly due to resorption of these elements during the decarbonation step. As most Zn was found to be present in a more difficult acid-extractable solid phase, its extractability accounted for only 25%. While reaction (pH) conditions of both decarbonation and solubilisation determined removal efficiencies, washing the extracted soil with deionized water only slightly increased the amount of metals removed. Metal distribution among solid phases — exchangeable, carbonate, reducible, organically bound, and residual — was affected by the different treatments. The amount of metals contained in the exchangeable and residual fractions determined their extractability. Except for Cu, the reducible and organically bound fractions were less important. After solubilisation 13 to 70% of the metals were present in an exchangeable solid phase. This implicates that washing the solubilized soil with a salt may increase the extractability of metals, especially for Zn and Pb. Based on our results the process is critically evaluated and possibilities for optimization formulated.     

Pilot-scale counter-current acid leaching process for Cu,Pb, Sb,and Zn from small-arms shooting range soil

Purpose

The main objective of this study was to evaluate the potential of a counter-current leaching process (CCLP) on 14 cycles with leachate treatment at the pilot scale for Pb, Cu, Sb, and Zn removal from the soil of a Canadian small-arms shooting range.

Materials and methods

The metal concentrations in the contaminated soil were 904?±?112 mg Cu kg^–1, 8,550?±?940 mg Pb kg^–1, 370?±?26 mg Sb kg^–1, and 169?±?14 mg Zn kg^–1. The CCLP includes three acid leaching steps (0.125 M H₂SO₄?+?4 M NaCl, pulp density (PD)?=?10 %, t?=?1 h, T?=?20 °C, total volume?=?20 L). The leachate treatment was performed using metal precipitation with a 5-M NaOH solution. The treated effluent was reused for the next metal leaching steps.

Results and discussion

The average metal removal yields were 80.9?±?2.3 % of Cu, 94.5?±?0.7 % of Pb, 51.1?±?4.8 % of Sb, and 43.9?±?3.9 % of Zn. Compared to a conventional leaching process, the CCLP allows a significant economy of water (24,500 L water per ton of soil), sulfuric acid (133 L H₂SO₄ t^–1), NaCl (6,310 kg NaCl t^–1), and NaOH (225 kg NaOH t^–1). This corresponds to 82 %, 65 %, 90 %, and 75 % of reduction, respectively. The Toxicity Characteristic Leaching Procedure test, which was applied on the remediated soil, demonstrated a large decrease of the lead availability (0.8 mg Pb L^–1) in comparison to the untreated soil (142 mg Pb L^–1). The estimated total cost of this soil remediation process is 267 US$ t^–1.

Conclusions

The CCLP process allows high removal yields for Pb and Cu and a significant reduction in water and chemical consumption. Further work should examine the extraction of Sb from small-arms shooting range.     

污泥加石灰后作为肥料用于玉米生长中施肥研究

The use of sewage sludge on agricultural land provides an alternative for its disposal. Therefore, the aim of the present study was to evaluate the feasibility of using industrial sewage sludge produced in Pakistan, as an agricultural fertilizer. The agricultural soil amended with 250 g kg^-1 sewage sludge with or without lime treatment was used for the growth of the common local grain crop, maize （Zea maize）. The mobility of the trace and toxic metals in the sludge samples was assessed by applying a modified BCR sequential extraction procedure. The single extraction procedure was comprised of the application of a mild extractant （CaCl2） and water, for the estimation of the proportion of easily soluble metal fractions. To check the precision of the analytical results, the concentrations of trace and toxic metals in every step of the sequential extraction procedure were summed up and compared with total metal concentrations. The plant-available metal contents, as indicated by the deionized water and 0.01 mol L^-1 CaCl2 solution extraction fractions and the exchangeable fraction of the sequential extraction, decreased significantly （P 〈 0.05） with lime application because of the reduced metal availability at a higher pH, except in the cases of Cd and Cu, whose mobility was slightly increased. Sludge amendment enhanced the dry weight yield of maize and the increase was more obvious for the soil with lime treatment. Liming the sewage sludge reduced the trace and toxic metal contents in the grain tissues, except Cu and Cd, which were below the permissible limits of these metals. The present experiment demonstrates that liming was an important factor in facilitating the growth of maize in sludge-amended soil.     

EDTA Extraction of Heavy Metals from Different Soil Fractions and Synthetic Soils

A laboratory-prepared contaminated soil was partitioned into four fractions, namely carbonate, Fe/Mn oxides, organic matter and clay mineral, according to the form in which the heavy metal bound with soil constituents. Individual contaminated soil fractions and synthetic soils were prepared for the study of soil extraction using ethylenediaminetetraacetic acid (EDTA). The effect of contact time and EDTA concentration were evaluated for both individual soil fractions and synthetic soils. The extraction reached equilibrium rapidly, after about 30 min. A 0.01 M EDTA solution was less effective than a 0.05 M or a 0.10 M EDTA. EDTA was proved to be effective for metal removal from the four individual soil fractions and synthetic soils. In general, approximately 90% of metals were removed from synthetic soils by 0.10 M EDTA. EDTA extraction of Pb from a contaminated carbonate fraction was thought to be affected by the formation of lead carbonates. A simple equation based on the sum of the released heavy metal from the individual components is used to check if there are interactions among the different soil components when mixed. The estimated values agreed well with the experimentally measured results only for the 0.10 M EDTA system.     

Determination of mobile heavy metal fraction in soil: Results of a pot experiment with sewage sludge

Abstract

The development of a method using a chelating resin to assess heavy metal mobility in soil and the first results obtained from a pot experiment with sewage sludge additions were studied. The resin was Chelex 100 with the calcium (Ca)‐form of the resin proving to be best suited for the extraction. The efficiency of recovery of the heavy metals from an aqueous solution ranged from 81.2% for cadmium (Cd) to 102% for copper (Cu) within 24 hours. For heavy metal extractions from a soil sample, a 96 hour extraction period was found to be optimum. The extracted heavy metal portion was comparable with the results obtained with an ammonium acetate (NH₄AOc) extraction. Total heavy metal contents in the substrate of the pot experiment did not show a significant influence due to the sewage sludge treatments, although considerable amounts of heavy metals were added by the sewage sludge. This effect can be both due to the incomplete recovery of heavy metals by an aqua regia extraction and leaching losses of these elements from the pots. Rape (Raphanus sativus L.) plants did not have any heavy metal contents which might indicate a high availability in soil, with the Cd and Cr contents in the rape biomass being partly lower in the sewage sludge‐treated pots than in the control plants; however, zinc (Zn) uptake slightly increased with increasing sewage sludge treatments. The Chelex 100 extraction procedure was correlated with Cd plant uptake, while the NH₄AOc extraction procedure was better related to the Zn uptake by rape plants.     

Remediation of lead-, arsenic-, and cesium-contaminated soil using consecutive washing enhanced with electro-kinetic field

Purpose

Extensive deposition of Pb, As, and Cs in soils may damage ecosystems and human’s health. Soil washing is the most conventional remediation method, and its efficiency depends on metal solubility in soil. This study aims to optimize operating variables of electro-kinetic field (EKF)-enhanced soil washing procedures.

Materials and methods

Soil samples from a Mississippi River Delta rice field were homogeneously spiked with Pb, As, and Cs, and contaminated soil was aged for 3 months. The remediation involved a first stage electro-kinetic process, followed by a soil washing procedure. Soil pH changes under EKF were studied. Effects of citric acid and reversed EKF were investigated for alleviating possible alkaline precipitation. In the washing procedure, soil washing time and cycles with different extractants were examined. The overall EKF-enhanced soil washing efficiencies were discussed as well.

Results and discussion

The implement of EKF offered an acidic soil environment around the anode areas for solubilizing metal(loid)s. Combined with EKF, citric acid was more conductive to desorb metal(loid)s. In addition, the reversed EKF effectively alleviated metal(loid) precipitation caused by alkalization in the first stage cathode areas. The EKF significantly enhanced metal(loid) extractions in the anode area of soils using Na₂EDTA, CaCl₂, and citric acid at pH of 2. The most preferable removal of Pb (80–98 %), As (48–63 %), and Cs (10–13 %) was achieved with three extractants. CaCl₂ and citric acid were proved to be suitable alternatives to Na₂EDTA for Pb extraction. A washing process of 2 h extraction with double washing cycles was optimized.

Conclusions

Soil washing time and cycles were major factors governing the metal(loid) removal from soil. Washing process of 2 h extraction with double cycles was optimized for further extraction based on higher washing efficiency. The EKF effectively improved washing efficiency while some electrical parameters need further studies for cost performance consideration.

     

An improved method for determination of adenosine triphosphate (ATP) in soil

Different methods for extraction of ATP from soil were examined. The methods were compared with respect to the efficiency of extraction of ATP from soil, and the content of ATP in the extracts was measured by the luciferin-luciferase system. The following extraction procedure was found to be the most efficient:Shaking of the soil with sulphuric acid, followed by filtration, cation exchange of the extract on Na⁺ resin, and adjustment to pH 9.8 with ethanolamine.The method is simpler, faster and more convenient for processing large numbers of samples, than the earlier-described acid extraction methods. Furthermore, ATP levels of 5ngATPg^?1 dry wt soil can be measured and results can be reproduced within ± 5%.The respiration rates of the soils were also measured, and a high degree of correlation was found between carbon dioxide production and ATP levels determined by extraction with sulphuric acid or dimethylsulfoxid.     

Extraction of light hydrocarbons from soil

The recovery of light hydrocarbon constituents from three soils using three kinds of extraction methods, conducted in duplicates, was evaluated. Higher molecular weight compounds were recovered at greater than 80% whereas the recoveries of benzene, toluene, ethylbenzene, and xylene were generally less than 50%. Using the most efficient extraction procedure, the average recovery of light hydrocarbon from a soil, was 78%. Increased soil moisture content decreased the extent of recovery. Methanol and 2-propanol showed similar extracting potential for light hydrocarbon from soil. The rate of hydrocarbon removal was faster when methanol was used as a solvent, but its efficiency was similar to 2-propanol. The batch and column extraction methods gave very similar results for both solvents. However, column extraction offered an advantage over the soxhlet and batch procedures by reaching equilibrium faster.     

Determination of free DNA in soils

The concentration of free DNA in soils has to be determined in order to understand the fate and the transport of extracellular DNA. A protocol for the extraction and determination of free DNA was developed. The procedure uses separation steps, i.e. centrifugation and ultra‐filtration. The free dsDNA was stained with PicoGreen® and determined fluorimetrically. Samples from different soils, different soil horizons, soil waters and under different land use systems were analyzed. It was found that in nearly all samples free DNA was detectable. Free DNA concentrations of up to 1950 ng (g dried sample)^—1 could be detected depending on depth, soil type and system of land use.     

Multi‐element stable isotopic dilution and multi‐surface modelling to assess the speciation and reactivity of cadmium and copper in soil

Chemical extraction, multi‐element stable isotopic dilution (ID) and multi‐surface modelling were used to investigate the lability of cadmium (Cd) and copper (Cu) in nine types of soil with different properties and contaminated or not with Cd and Cu. The chemical extraction and ID analyses both showed that Cd was more labile than Cu in all the soil types studied. From the ID results, 32.8–93.3% of total Cd and 14.7–71.8% of total Cu were isotopically exchangeable after 3 days of equilibration. A single extraction in 0.43 m HNO₃ gave similar results to the 3‐day ID assay for Cu in most of the soils and for Cd in the non‐calcareous soils. However, an eight‐step selective sequential extraction (SSE) procedure gave different results from the ID assay for both metals. Predictions of the multi‐surface model for the amounts of Cd and Cu adsorbed, based on measured metal ion activities in the soil solution and the concentrations of reactive surfaces in the soil, agreed with the ID results. The model predicted that soil organic matter was the predominant sorbent for Cd and Cu in the soils and that manganese oxide was the least important sorbent. The contributions of iron oxides to sorption were predicted to be small except in soil with a high pH and little organic matter. The predicted sorption on different soil components did not match SSE measurements.     

Distribution of Chemical Forms for Co, Cr, Ni and V in Typical Soils of China

Co,Ni,Cr and V in 25 typical soils of China were fractionated into exchangeable,carbonate bound (calcareous soils),Mn oxide bound,organically bound,amorphous Fe oxide bound,crystalline Fe oxide bound and residual forms using a seven-step sequential extraction procedure,so as to study the distribution of chemical forms of these metals as well as the effects of soil proiperties on the distribution.The results showed that most of soil Co,Ni,Cr and V were present in residual forms,and the distribution ratio averaged 48.2% for Co,53.0% for Ni,81.5% for Cr and 68.7% for V.The speciation of heavy metals was greatly influenced by soil physico-chemical properties and the chemistry of elements.The results also indicated that the recovery of metal elements by the sequential extraction procedure was satisfactory,with the relative difference between the sum of seven forms and the total content in soils averaging 9.5% for Co,12.8% for Ni,6.6% for Cr and 7.2% for V.     

Ultrasound Method for Extraction of Phosphorus in Soil

In the present work, an ultrasoundic procedure for the extraction of phosphorus (P) available in soil is described. The proposed method is based on extraction by 0.5 M sodium bicarbonate (NaHCO₃), following with sonication under different conditions. Phosphorus was determined by the Murphy and Riley method. Sonication time and soil–extractant relative quantities were optimized. A statistical analysis approach was used to find suitable conditions for the ultrasoundic extraction. The main advantages of the sonication method are the reduced times of extractions, which take 10 min in contrast to the 30 min required by a shaking method, and the possibility to reduce soil and extractant quantities from 5 g–100 mL to 2 g–40 mL. Performance of the method was evaluated, and the procedure was utilized to analyze soils from Santiago del Estero, Argentina.     

Soil Extraction of Readily Soluble Heavy Metals and As with 1 M NH4NO3-Solution - Evaluation of DIN 19730 (6 pp)

Background, Aims and Scope   The German DIN 19730 (1997) describes a method for the extraction of readily available trace elements from soil by shaking the soil with 1 M NH4NO3-solution. Based on this method trigger and action values for the transfer of heavy metals and arsenic from soils to plants have been published in the German Federal Soil Protection and Contaminated Sites Ordinance (BBodSchV 1999). The chemical mechanisms involved in this soil extraction procedure were evaluated in some detail to create requirements to improve environmental risk assessment for soil contaminations.Methods   The chemical mechanisms involved when soil is extracted with 1 M NH4NO3-solution were evaluated. This was followed by a laboratory experiment to quantify the formation of soluble metal ammine complexes during the extraction. Cd, Zn, Ni, Cu, Co and Hg were extracted from 16 soils by 1 M NH4NO3, 1 M KNO3 and water. pH was adjusted in 5 steps between 5.0 to 7.5. The potassium cation (K+) and the ammonium cation (NH4+) behave similarly when cations from soil surfaces are desorbed, because they have almost identical ion radii (e.g. 0.133 and 0.143 nm). K+ does not form ammine complexes with other ions, whereas, due to the increasing formation of NH3 from NH4 by dissociation with rising pH, metal ammine complex formation is an important process in soil extraction when using ammonium salt solutions. A difference in the extraction efficiency of 1 M NH4NO3- and 1 M KNO3-solution for a given soil can therefore be attributed to the formation of soluble metal ammine complexes. Conclusion   Our experiments resulted in considerably higher extraction rates of Cu, Cd and Hg by 1 M NH4NO3-solution as compared to 1 M KNO3-solution. This effect, caused by the formation of soluble metal ammine complexes, was only evident in soils with higher readily soluble heavy metal contents and a soil pH above 6 – 6.5 for Cu and 7 – 7.5 for Cd. Further chemical mechanisms involved when soils are extracted with 1 M NH4NO3 are a moderate decrease in pH and an increase in ionic strength. Most of the colloids and parts of soluble metal-organic complexes are precipitated due to the high ionic strength. High ionic strength also decreases the activity of metal-OH+ species and the electrostatic potential of the particle surfaces, which in turn, increases the desorption of heavy metal cations from negatively charged soil surfaces. In contrast, the adsorption of anions like arsenate is favoured by the decreasing electrostatic potential. The prediction of heavy metal uptake by plants from the results of the 1 M NH4NO3-solution extraction fits well for elements, which are mainly bound by low strength electrostatic forces to the soils. Such conditions are found in acidic soils for Cd and Tl, which have a low tendency for hydrolysis compared to other heavy metals. The correlation between 1 M NH4NO3 soil extraction and plant uptake is less significant for Ni and Zn. Only low positive correlation coefficients have been found for Pb, As, Hg and for the Cu-uptake by wheat. Imprecise prediction of plant uptake of heavy metals by the extraction with 1 M NH4NO3-solution is mainly caused by conditions leading to an overestimation of plant availability such as elements are strongly bound to soils, or low soluble trace element contents in soils. Neutral to alkaline soil pH can also lead to imprecise prediction due to increasing formation of soluble metal-organic (Cu, Pb, Hg) and metal ammine (Hg, Cu, Cd) complexes and less importantly due to the formation of colloids. Therefore, at low 1 M NH4NO3-extractable soil contents usually no high plant contents are to be expected. Recommendation and Outlook   Extraction of soil with 1 M NH4NO3-solution is a suitable method for the determination of readily soluble and plant available trace element contents. The chemical soil extraction process may cause misleading predictions of the transfer of trace elements to plants for some soil properties. This knowledge should be used to improve risk assessment of soil contaminations. It has to be considered, that the processes involved in plant uptake of trace elements are too complex to expect that just one soil extraction method can always guarantee a correct prognosis of toxicological significant element contents in plants. Soil analyses may be used for the preliminary examination of suspicious areas and the demarcation of contaminated areas. The results of soil analyses should be checked additionally by plant analyses especially under conditions with a high probability for misleading results by the 1 M NH4NO3-extraction. Alternatively, extraction with 1 M KNO3-solution can be performed to exclude the effect of metal ammine complex formation.     

Small Scale Heterogeneity of Soil Chemical Properties. II. Fractions of Aluminum and Heavy Metals

Soil aggregates from nine horizons of five forest sites in Bavaria were mechanically separated into an aggregate surface fraction and an aggregate core fraction. Seven operationally defined fractions of AI, Cd, Cu, Cr, Ni, Pb, and Zn were determined by a sequential extraction procedure within both aggregate fractions. The CEC, C_org, and oxides of Mn and Fe also were measured. The differences in metal concentrations between surface and core fractions were small but consistent: mainly lithogenic metals (Al, and, within a serpentine derived soil, Ni and Cr) showed lower total concentrations in the surface than in the core fraction. Metals originating primarily from atmospheric deposition form two groups. Cadmium, Pb, and Zn usually showed higher total concentrations in the surface fraction whereas Cu, Cr, and Ni showed no uniform trends. In general, lower proportions of the total metal concentration could be extracted by the residual extraction step (mainly bound within silicates) in the surface than in the core fraction. Accordingly, higher proportions could be extracted by the sum of all other steps (bound to cation exchangers, organic matter, and oxides) in the surface fraction, although in the aggregate surface fraction the concentrations of organic matter and oxides as well as the CEC were lower than in the core fraction. The fact that the sorption capacity of the surface is lower than of the core fraction has to be considered for the identification of external metal inputs. Based on these results, a new method to identify external inputs is proposed.     

EDTA Leaching of Cu Contaminated Soils Using Ozone/UV for Treatment and Reuse of Washing Solution in a Closed Loop

Ozone and UV irradiation were used for oxidative decomposition of EDTA-Cu complexes in washing solution obtained during multi-step leaching of Cu (344,1?±?36.5 mg kg^?1) contaminated vineyard soil with EDTA as a chelant. The released Cu was absorbed from the washing solution on a commercial mixture of metal absorbing minerals, and the treated washing solution then reused for removal of soil residual Cu-EDTA complexes in a closed-loop process. Six consecutive leaching steps (6?×?2.5 mmol kg^?1 of EDTA) removed 38.8 % of Cu from soils, and reduced Cu soil mobility, determined using the toxicity characteristic leaching test (TCLP), by 28.5%. The final washing solution obtained after soil remediation was colourless, with a pH close to neutral (7.5?±?0.2) and with low concentrations of Cu and EDTA (0.51?±?0.22 mg L^?1 and 0.083 mM, respectively). The proposed remediation method has therefore potential not just to recycle and save process water, but also not to produce toxic wastewaters. Soil treatment did not substantially alter the soil properties determined by pedological analysis, and had relatively little impact on soil hydraulic conductivity and soil water sorption capacity.     

An overview of fertilizer‐P recommendations in Europe: soil testing,calibration and fertilizer recommendations

The procedure for applying phosphorus (P) fertilizer to soil can be divided into three consecutive steps: (i) Measurement of soil‐P availability, (ii) calibration of the soil‐P fertility level and (iii) estimation of the recommended P dose. Information on each of these steps was obtained for 18 European countries and regions with the aim of comparing P fertilizer recommendation systems at the European scale. We collected information on P fertilizer recommendations through conventional or grey literature, and personal contacts with researchers, laboratories and advisory services. We found much variation between countries for each of the three steps: There are more than 10 soil‐P tests currently in use, apparent contradictions in the interpretation of soil‐P test values and more than 3‐fold differences in the P fertilizer recommendations for similar soil‐crop situations. This last result was confirmed by conducting a simple experimental inter‐laboratory comparison. Moreover, soil properties (pH, clay content) and crop species characteristics (P responsiveness) are used in some countries in the calibration and recommendation steps, but in different ways. However, there are also common characteristics: soil‐P availability is determined in all countries by extraction with chemical reagents and the calibration of the soil‐P test values, and the fertilizer recommendations are based on the results from empirical field trials. Moreover, the fertilizer recommendations are nearly all based on the amount of P exported in the crops. As long as rational scientific and theoretical backgrounds are lacking, there is no point in trying to synchronize the different chemical methods used. We therefore call for a mechanistic approach in which the processes involved in plant P nutrition are truly reproduced by a single standard method or simulated by sorption‐desorption models.     

Use of the BCR three-step sequential extraction procedure for the study of the partitioning of Cd, Pb and Zn in various soil samples

A slightly modified three-step sequential extraction procedure proposed by the Community Bureau of Reference (BCR) for analysis of sediments was successfully applied to soil samples. Contaminated soil samples from the lead and zinc mining area in the Mezica valley (Slovenia) and natural soils from a non-industrial area were analysed. The total concentrations of Cd, Pb and Zn and their concentrations in fractions after extraction were determined by flame or electrothermal atomic absorption spectrometry (FAAS, ETAAS). Total metal concentrations in natural soils ranged from 0.3 to 2.6 mg kg^-1 for Cd, from 20 to 45 mg kg^-1 for Pb and from 70 to 140 mg kg^-1 for Zn, while these concentrations ranged from 0.5 to 35 mg kg^-1 for Cd, from 200 to 10000 mg kg^-1 for Pb and from 140 to 1500 mg kg^-1 for Zn in soils from contaminated areas. The results of the partitioning study applying the slightly modified BCR three-step extraction procedure indicate that Cd, Pb and Zn in natural soils prevails mostly in sparingly soluble fractions. Cd in natural soils is bound mainly to Fe and Mn oxides and hydroxides, Pb to organic matter, sulphides and silicates, while Zn is predominantly bound to silicates. In contaminated soils, Cd, Pb and Zn are distributed between the easily and sparingly soluble fractions. Due to the high total Cd, Pb and Zn concentrations in contaminated soil close to the smelter, ! and their high proportions in the easily soluble fraction (80% of Cd, 50% of Pb and 70% of Zn), the soil around smelters represents an environmental hazard.     

三种无机萃取剂去除土壤中PbCd的条件研究

通过对重金属污染土壤的萃取实验,比较了HCl、Na2S2O3、CaCl2 3种常用无机萃取剂对重金属Ph和cd的萃取效果,并通过改变萃取条件（萃取剂浓度、时间、固液比、萃取次数和温度）,以期找到各种萃取剂达到最大萃取效率时所需的萃取条件。结果表明,萃取剂浓度对于萃取效率的影响最大,随着浓度的提高,3种萃取剂对Ph、Cd的去除作用增强;固液比和萃取次数对Ph、Cd的去除率也有较大的影响;时间对两金属的去除率也有一定的影响,但在本实验条件下,达到萃取平衡所需时间很短;温度对萃取效率的影响很小。本实验中,HCl去除Pb、Cd的能力远远大于其他两种萃取剂。对Cd进行萃取时,需增加萃取次数才能达到较好的萃取效果。     

Impact of DNA extraction method on bacterial community composition measured by denaturing gradient gel electrophoresis

The impact of DNA extraction protocol on soil DNA yield and bacterial community composition was evaluated. Three different procedures to physically disrupt cells were compared: sonication, grinding-freezing-thawing, and bead beating. The three protocols were applied to three different topsoils. For all soils, we found that each DNA extraction method resulted in unique community patterns as measured by denaturing gradient gel electrophoresis. This indicates the importance of the DNA extraction protocol on data for evaluating soil bacterial diversity. Consistently, the bead-beating procedure gave rise to the highest number of DNA bands, indicating the highest number of bacterial species. Supplementing the bead-beating procedure with additional cell-rupture steps generally did not change the bacterial community profile. The same consistency was not observed when evaluating the efficiency of the different methods on soil DNA yield. This parameter depended on soil type. The DNA size was of highest molecular weight with the sonication and grinding-freezing-thawing procedures (approx. 20 kb). In contrast, the inclusion of bead beating resulted in more sheared DNA (approx. 6-20 kb), and the longer the bead-beating time, the higher the fraction of low-molecular weight DNA. Clearly, the choice of DNA extraction protocol depends on soil type. We found, however, that for the analysis of indigenous soil bacterial communities the bead-beating procedure was appropriate because it is fast, reproducible, and gives very pure DNA of relatively high molecular weight. And very importantly, with this protocol the highest soil bacterial diversity was obtained. We believe that the choice of DNA extraction protocol will influence not only the determined phylogenetic diversity of indigenous microbial communities, but also the obtained functional diversity. This means that the detected presence of a functional gene—and thus the indication of enzyme activity—may depend on the nature of the applied DNA extraction procedure.     

施用碱稳定固体的酸性土壤的Cu和Zn的形态分布

Fractionation of metals in a granite-derived acid sandy loam soil amended with alkaline-stabilised sewagesIudge biosolids was conducted in order to assess metal bioavailability and environmental mobility soil solution was extracted by a centrifugation and filtration technique. Metal speciation in the soil solution wasdetermined by a cation exchange resin method. Acetic acid and EDTA extracting solutions were used forextraction of metals in soil solid surfaces. Metal distribution in different fractions of soil solid phase was determined using a three-step sequential extraction scheme. The results show that the metals in the soilsolution existed in different fractions with variable lability and metals in the soil solid phase were also presentin various chemical forms with potentially different bioavail ability and environmental mobility Alkaline-stabilised biosolids could elevate solubility of Cu and proportion of Cu in organically complexed fractionsboth in soil liquid and solid phases, and may therefore increase Cu mobility. In contrast, the biosolids lowered the concentrations of water-soluble Zn (labile fraction) and exchangeable Zn and may hence decrease bioavailability and mobility of Zn. However, Fe and Mn oxides bound and organic matter bound fractions are likely to be Zn pools in the sludge-amended soil. These consequences possibly result from the liming effect and metal speciation of the sludge product and the difference in the chemistry between the metals in soil.