Sečovlje salterns are an important protected area of biotic diversity in the Mediterranean. They represent an extreme environment with high salinity and drought that severely influence the growth of ...organisms. In the present study, diversity of plant halophytes and their mycorrhizal status were screened at eight different locations, which were mostly dikes and salt ponds, and which were deliberately selected for their distinct properties (e.g. soil salinity ranging from 105 to 2627 μS cm
−1, vegetation type and management practice of the salterns).
Twelve different halophytic plant species were recorded, of which eleven are designated as vulnerable. With few exceptions, they were found at the abandoned (Fontanigge) and sustained (Lera) locations of the Sečovlje salterns, distributed according to their tolerance to the salinity and waterlogging. The highest diversity of halophytes was listed at Fontanigge, in the abandoned, periodically flooded and gradually overgrown salt ponds. All of the examined species were colonised with either arbuscular mycorrhizal fungi (AMF) and/or dark septate endophytes (DSEs). High levels of colonisation were however detected only for species belonging to the Asteraceae and Plantaginaceae families. Higher root colonisation frequencies were generally seen for plants growing in the abandoned parts, when compared to the managed parts, whereas there was little correlation of the colonisation parameters with physicochemical parameters of rhizospheric soil properties.
Molecular analysis by temporal temperature gradient gel electrophoresis (TTGE) of roots of halophytic plant species with confirmed AMF colonisation (arbuscules present) revealed the occurrence of at least six different AMF species, related to
Glomus geosporum,
Glomus caledonium and
Glomus intraradices, and to different
Glomus sp. clades and the
Diversispora clade. This is to the best of our knowledge the first report of AMF and DSE mycorrhizal status of most of the halophyte plant species examined and of the brother scale identification of AMF species based on molecular analyses of roots of diverse halophytes from high saline environments.
The effect of two earthworm species,
Lumbricus rubellus and
Eisenia fetida, on the fractionation/bioavailability of Pb and Zn before and after soil leaching with EDTA was studied. Four leaching steps ...with total 12.5
mmol
kg
−1 EDTA removed 39.8% and 6.1% of Pb and Zn, respectively. EDTA removed Pb from all soil fractions fairly uniformly (assessed using sequential extractions). Zn was mostly present in the chemically inert residual soil fraction, which explains its poor removal. Analysis of earthworm casts and the remainder of the soil indicated that
L. rubellus and
E. fetida actively regulated soil pH, but did not significantly change Pb and Zn fractionation in non-remediated and remediated soil. However, the bioavailability of Pb (assessed using Ruby's physiologically based extraction test) in
E. fetida casts was significantly higher than in the bulk of the soil. In remediated soil the Pb bioavailability in the simulated stomach phase increased by 5.1 times.
Earthworm activity increases heavy metal bioavailability in soil before and after remediation.
Pet reprezentančnih vzorcev močno onesnažene zemljine z območja stare cinkarne v Celju (Slovenija), ki so vsebovali od 7300 do 17200 mg kg-1 celokupnega Pb, od 6000 do 63600 mg kg-1 celokupnega Zn in ...od 24 do 250 mg kg-1 celokupnega Cd, smo remedirali z metodo stabilizacije s 15 ut.% portland cementa. Po 4 mesecih stabilizacije se je mobilnost Pb, Zn in Cd določena s TCLP metodo ("Toxicity Characteristic Leaching Procedure") 5,8 - 17,4, 8,1 - 35,8 oz. 4,3 - 7,3-krat zmanjšala. Potencialna dostopnost Pb, Zn in Cd za rastline, določena z ekstrakcijo z dietilentriaminpentaocetno kislino (DTPA), se je po stabilizaciji 1,6 - 9,8, 1,1 - 2,1 oz. 1,3 - 4,1-krat zmanjšala. Biodostopni delež Pb določen s fiziološko osnovanim ekstrakcijskim testom ("Physiologically Based Extraction Test"; PBET) se je v črevesni frakciji po stabilizaciji 5,9 - 11,3-krat zmanjšal.
► Leaching with EDTA and HCl removes PTMs and decreases their accessibility. ► EDTA is a more efficient leachant compared to HCl. ► EDTA increased the pH of the acidic soil. ► HCl significantly ...dissolved carbonates from calcareous soil. ► HCl had a negative impact on soil microbial and enzyme activity in both soils.
The environmental risk of potentially toxic metals (PTMs) in soil can be diminished by their removal. Among the available remediation techniques, soil leaching with various solutions is one of the most effective but data about the impact on soil chemical and biological properties are still scarce. We studied the effect of two common leaching agents, hydrochloric acid (HCl) and a chelating agent (EDTA) on Pb, Zn, Cd removal and accessibility and on physico-chemical and biological properties in one calcareous, pH neutral soil and one non-calcareous acidic soil. EDTA was a more efficient leachant compared to HCl: up to 133-times lower chelant concentration was needed for the same percentage (35%) of Pb removal. EDTA and HCl concentrations with similar PTM removal efficiency decreased PTM accessibility in both soils but had different impacts on soil properties. As expected, HCl significantly dissolved carbonates from calcareous soil, while EDTA leaching increased the pH of the acidic soil. Enzyme activity assays showed that leaching with HCl had a distinctly negative impact on soil microbial and enzyme activity, while leaching with EDTA had less impact. Our results emphasize the importance of considering the ecological impact of remediation processes on soil in addition to the capacity for PTM removal.
Ozone and UV irradiation were used for oxidative decomposition of EDTA-Cu complexes in washing solution obtained during multi-step leaching of Cu (344,1c36.5mg kg super(-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 (62.5mmol kg super(-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 (7.5c0.2) and with low concentrations of Cu and EDTA (0.51c0.22mg L super(-1) and 0.083mM, 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.
Soil washing remediation techniques usually remove only the labile heavy metal (HM) species from the soil, leaving the residual ones in less available/mobile forms, thus disturbing the chemical ...equilibrium among different species of HM in the soil. Re-establishing such equilibrium and shifting HM back to more available/mobile chemical forms could occur after exposing the remediated soil to environmental abiotic (ageing) factors. Contaminated soil from a smelter site (Pb 4600mgkg−1, Zn 1800mgkg−1, Cd 30mgkg−1) was leached with increasing EDTA concentrations (2.5, 5.0, 10.0, 20.0, 40.0 and 4-consecutive steps of 40.0mmolEDTAkg−1 of soil). A gradient of removed HM was reached: from 6% to 73% of initial Pb, from 3% to 23% of initial Zn and from 17% to 74% of initial Cd were removed. Repetitive temperature changes (105°C and −20°C) were used to mimic abiotic factors acting on residual HM after EDTA soil leaching in saturated soil at 10% and 90% of soil water holding capacity. Fractionation using sequential extractions, mobility, and phytoavailability of Pb, Zn and Cd and Pb oral bioavailability were determined for aged and non-aged soil. The ageing treatment consistently lowered HM phytoavailability in the original (non-leached) and all treated (chelant-leached) soils. However, Pb, Zn and Cd behaved differently from each other; Pb mobility increased, Cd mobility decreased, while Zn mobility did not change. The results indicate that abiotic (ageing) processes change the availability/mobility of residual HM in all leaching treatments and should thus be considered in final remediation effectivity evaluation.
To reveal their mycotrophic status, we have here analysed the fungal colonisation of the roots, and the identity of these fungi, of the most abundant halophytic plant species from Sečovlje salterns ...(Slovenia): Aster tripolium, Limonium angustifolium and Salicornia europaea. The highest frequency of fungal colonisation was seen for the roots of A. tripolium, followed by those of L. angustifolium and S. europaea. Hyphae and occasional microsclerotia, the assumed structures of dark septate endophytes, were seen in the roots of all three species, whereas arbuscules, as typical structures of arbuscular mycorrhiza, were seen in the roots of A. tripolium and in one specimen of L. angustifolium. Fungal partial small subunit ribosomal DNA (SSU-rDNA) fragments were amplifi ed from total root DNA extracts for further restriction fragment length polymorphism (RFLP) analyses of the structures of the fungal root communities. Fifteen different RFLP profi les were obtained that grouped into two major clusters. Two RFLP profi les strongly dominated in samples of all three of the plant species. Sequencing revealed that one of these profiles corresponds to the species Cylindrobasidium laeve (Basidiomycota), and the second to Capnobotryella sp. / Phaeotheca fi ssurella, putative dark septate endophytes from the class Dothideomycetes (Ascomycota). Intraspecific RFLP polymorphisms were demonstrated for both species. To our knowledge, this is the first report on dark septate endophyte fungi occurrence in roots of A. tripolium, L. angustifolium and S. europaea.
Remediation by means of soil leaching with ethylenediaminetetraacetic acid (EDTA) is capable of extracting the most labile soil fractions, leaving the residual metals in biologically non-available ...forms. We evaluated the feasibility of the standardized earthworm (
Eisenia fetida) avoidance test for assessing the efficiency of soil remediation of Pb, Zn and Cd polluted soil. Chemical extraction tests (six-step sequential extraction, toxicity characteristic leaching procedure, physiologically based extraction test, diethylenediaminepentaacetic acid extraction) indicated that the mobility, oral bioaccessibility and phytoavailability of Pb, Zn and Cd were consistently reduced. However, the avoidance test showed no significant avoidance of polluted soil in favor of that which had been remediated. Pb, Zn and Cd accumulation in
E. fetida mirrored the decreasing pattern of metal potential bioavailability gained by leaching the soil with increasing EDTA concentrations. The calculated bioaccumulation factors indicated the possibility of underestimating the metal bioavailability in soil using chemical extraction tests.
Tests with indicator organisms should be used for a more meaningful and holistic assessment of metal biological availability in polluted and remediated soil.
Sečovlje salterns are an important protected area of biotic diversity in the Mediterranean. They represent an extreme environment with high salinity and drought that severely influence the growth of ...organisms. In the present study, diversity of plant halophytes and their mycorrhizal status were screened at eight different locations, which were mostly dikes and salt ponds, and which were deliberately selected for their distinct properties (e.g. soil salinity ranging from 105 to 2627 μS cm-1, vegetation type and management practice of the salterns). Twelve different halophytic plant species were recorded, of which eleven are designated as vulnerable. With few exceptions, they were found at the abandoned (Fontanigge) and sustained (Lera) locations of the Sečovlje salterns, distributed according to their tolerance to the salinity and waterlogging. The highest diversity of halophytes was listed at Fontanigge, in the abandoned, periodically flooded and gradually overgrown salt ponds. All of the examined species were colonised with either arbuscular mycorrhizal fungi (AMF) and/or dark septate endophytes (DSEs). High levels of colonisation were however detected only for species belonging to the Asteraceae and Plantaginaceae families. Higher root colonisation frequencies were generally seen for plants growing in the abandoned parts, when compared to the managed parts, whereas there was little correlation of the colonisation parameters with physicochemical parameters of rhizospheric soil properties. Molecular analysis by temporal temperature gradient gel electrophoresis (TTGE) of roots of halophytic plant species with confirmed AMF colonisation (arbuscules present) revealed the occurrence of at least six different AMF species, related to Glomus geosporum, Glomus caledonium and Glomus intraradices, and to different Glomus sp. clades and the Diversispora clade. This is to the best of our knowledge the first report of AMF and DSE mycorrhizal status of most of the halophyte plant species examined and of the brother scale identification of AMF species based on molecular analyses of roots of diverse halophytes from high saline environments.
Background, aim, and scope 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. Materials and methods We used soil from a 50-year-old vineyard regularly managed and treated with CuSO₄•5H₂O (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. Discussion 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. Recommendations and perspectives 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.
