title: Fractionation and bioavailability of Cu in soil remediated by EDTA leaching and processed by earthworms (Lumbricus terrestris L.).
abstract: OBJECTIVE
  Soil remediation with ethylenediamine tetraacetic acid (EDTA) leaching is capable of removing only part of the total metal concentration in the soil, mostly the labile, bioavailable metal species (metal bioavailability stripping). However, reintroduction of remediated soil in the environment exposes the soil to various environmental factors, which could potentially shift nonlabile residual metals back to labile bioavailable forms. We studied the effect of autochthonous earthworm species as model biotic environmental factor on the fractionation and bioavailability of Cu residual in soil after remediation.
  
  
  METHODS
  We used soil from a 50-year-old vineyard regularly managed and treated with CuSO(4)*5H(2)O (Bordeaux mixture) as fungicide. Soil containing 400 mg kg(-1) of Cu was leached with total 15 mmol kg(-1) EDTA. Remediated and nonremediated soil was processed by fully clitellated adult specimens of Lumbricus terrestris L., a prevailing autochthonous soil earthworm species. Cu fractionation, phytoavailability, and oral-bioavailability in processed and nonprocessed soil were determined using six-step sequential extraction, extraction with diethylenediamine pentaacetic acid, and in vitro physiologically based extraction test, respectively.
  
  
  RESULTS
  EDTA leaching removed 41% of the pseudototal Cu, mostly from the soil Fe- and Mn-oxides, carbonates, and organic matter. A 2.7-fold decrease in Cu phytoavailability and a 4.4- and 2.8-fold decrease in Cu oral-bioavailability in the stomach and small intestine fractions, respectively, were achieved after remediation. In nonremediated soil, earthworms increased the share of nonlabile Cu in residual soil fraction, while in remediated soil they increased the share of Cu bound to carbonates. A statistically significant 1.1- and 1.7-fold increase in Cu phytoavailability and intestinal oral-bioavailability, respectively, was observed in earthworm processed remediated soil.
  
  
  CONCLUSIONS
  Cu occurs in various soil "pools" of different solubilities with different chemical characteristics and consequently different functions. By removing the labile part of the metals from the soil during remediation, we disrupt the chemical equilibrium; the nonlabile residual metals left in soil after remediation might become more labile in time in tendency to re-establish that equilibrium. Earthworms alter the physical and chemical properties of soil affecting consequently the fractionation of metals. The increase in earthworm's gut pH due to the excretion of ammonia and/or calcium carbonate into the intestine could lead to the transbounding of metals into the carbonate fraction. However, their activity in remediated soil increased Cu phytoavailability and intestinal oral-bioavailability, and it would, therefore, be improper to generalize the influence of earthworms on metal availability in soil.
  
  
  CONCLUSIONS
  The results presented here show that residual Cu in remediated soil is affected by environmental factors such as earthworms, which should be considered in evaluating the effect of Cu polluted soil remediation.
  
  
  CONCLUSIONS
  Information on the behavior of residual metals in soil after its remediation is surprisingly scarce. The development of new effective remediation techniques should imply also the evaluation of postremediation effects on remediated soil. The results presented in this work indicate a possible tool for assessing the effect of biotic environmental factors on residual metals left in soil after its remediation.
authors.affiliation: Biotechnical Faculty, Agronomy Department, Centre for Soil and Environmental Science, University of Ljubljana, Ljubljana, Slovenia.
authors.forename: Metka
authors.initials: M
authors.lastname: Udovic
authors.forename: Domen
authors.initials: D
authors.lastname: Lestan
mesh_terms: D000818:Animals; D001673:Biodegradation, Environmental; D005591:Chemical Fractionation / Q000379:methods; D003300:Copper / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D004492:Edetic Acid / Q000737:chemistry*; D007700:Kinetics; D009835:Oligochaeta / Q000378:metabolism*; D012989:Soil Pollutants / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism
chemical_list: D012989:Soil Pollutants; D003300:Copper; D004492:Edetic Acid