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Phytomining of nickel (Ni) refers to cropping of selected Ni hyperaccumulator plants on Ni-rich serpentine soils. In this study, the effect of different fertilization regimes on the Ni yield of Odo...

AimsMetal hyperaccumulation is a rare phenomenon described for an increasing number of plant taxa. In this study we investigated the root growth responses of the well-known nickel, zinc, cadmium hyperaccumulatorNoccaea caerulescensand of the metal tolerant (non-accumulator)Stellaria media, in order to observe root foraging vs avoidance responses to nickel. MethodsTo allow for observations of root growth and foraging preferences, two accessions ofNoccaea caerulescensand two accessions ofStellaria mediaorginating from high nickel and low nickel habitats were grown in rhizotrons with localized nickel enrichment.ResultsThe root density in the control and nickel-enriched soil areas in the rhizotrons with differentN. caerulescensaccessions had distinct responses: moderate nickel avoidance was recorded for the non-nickel accession, while a clear foraging response was observed inN. caerulescensfrom the nickel accession. In contrast, nickel rooting avoidance was observed for bothS. mediaaccessions and was more pronounced in the non-nickel accession.ConclusionsThis study shows thatN. caerulescensoriginating from different accessions responded differently to soil nickel enrichment, with the nickel accession ofN. caerulescensactively foraging for nickel, suggesting a physiological adaptation and demand for this metal. In contrast, a clear nickel avoidance response by a metal tolerant species, S. media, was observed in this study, a phenomenon which has not been previously described; this suggests that root avoidance responses might play a role in the adaptation of metal tolerant species to Ni-rich soils.

Phytomining of nickel (Ni) is based on the cropping of Ni hyperaccumulators on Ni-rich serpentine soils. The efficiency of this approach is dependent on shoot nickel concentration and harvestable biomass. In a field experiment conducted on an Austrian serpentine site, the phytomining efficiency of the two plant species Odontarrhena chalcidica (syn. Alyssum murale) and Noccaea goesingensis was evaluated. O. chalcidica was planted in three treatments: control, sulphur application (0.46 g S kg−1 soil) and intercropping with the legume Lotus corniculatus. For N. goesingensis the treatments control, high-density planting (110 plants m−2) and intercropping were implemented. Given the experimental set-up, shoot biomass, shoot Ni concentration and thus the total amount of harvested Ni were on average higher for O. chalcidica. The highest Ni yield was achieved with O. chalcidica, reaching 55 kg Ni ha−1 in the sulphur treatment. N. goesingensis showed the maximum yield in the high-density treatment with 36 kg Ni ha−1. However, high-density planting of N. goesingensis and sulphur application to O. chalcidica plots did not significantly increase the Ni yield compared to the control. Intercropping with L. corniculatus tended to decrease the shoot biomass of both species. Planting of the hyperaccumulators led to a decrease of DTPA-extractable Ni and to an increase of soil pH, with the exception of sulphur-amended plots. Likewise, rhizosphere soil pH was higher than bulk soil values. Our data suggest that in particular O. chalcidica is suitable for Ni phytomining on the tested site. Measures to further increase the Ni yield and to optimise crop management will be evaluated in follow-up experiments.

Petroleum contamination and its remediation via plant-based solutions have got increasing attention by environmental scientists and engineers. In the current study, the physiological and growth responses of two diesel-tolerant plant species (tolerance limit: 1500-2000 mg/kg), Italian ryegrass (Lolium multiflorum) and Birdsfoot trefoil (Lotus corniculatus), have been investigated in vegetable oil- and diesel oil-amended soils. A long-term (147-day) greenhouse pot experiment was conducted to differentiate the main focus of the study: physical and chemical effects of oil (vegetable and diesel) in freshly spiked soils via evaluating the plant performance and hydrocarbon degradation. Moreover, plant performance was evaluated in terms of seed germination, plant shoot biomass, physiological parameters, and root biomass. Addition of both diesel oil and vegetable oil in freshly spiked soils showed deleterious effects on seedling emergence, root/shoot biomass, and chlorophyll content of grass and legume plants. Italian ryegrass showed more sensitivity in terms of germination rate to both vegetable and diesel oil as compared to non-contaminated soils while Birdsfoot trefoil reduced the germination rate only in diesel oil-impacted soils. The results of the current study suggest that both physical and chemical effects of oil pose negative effects of plant growth and root development. This observation may explain the phenomenon of reduced plant growth in aged/weathered contaminated soils during rhizoremediation experiments.

While the Galapagos Islands have been renowned for their unique flora and fauna since the time of Charles Darwin, the soils of the isolated island chain have been mostly overlooked and little information on their heavy metal contents is available. The aim of this study was therefore to examine the total heavy metal (Cd, Co, Cr, Cu, Ni, Pb, U, Zn) contents of soils from the agricultural areas on islands Isabela, Santa Cruz and San Cristobal, and identify trends with duration of exposure to weathering processes. Additionally, the mobility of these elements was assessed using ammonium nitrate extraction. In general, levels of Cd, Co, Cr, Cu, Ni and Zn were high compared to other world locations, while Pb levels were low and U levels were similar. Ni, Co, Cr, and to a lesser extent Pb and U tended to accumulate with increasing weathering duration. Soil concentrations of Cd, Zn, Cu, and possibly Pb and U, may have been influenced by use of agrochemicals, particularly on Santa Cruz Island. Mobility of Cd displayed an increasing trend with soil age, while Ni mobility decreased. Many soils had total contents of Cd, Co, Cr, Cu, Ni and Zn above threshold values indicating possible ecological or health risks. Systematic examination of trace element contents in soils from pristine national park areas would further assist in the delineation of background levels and the development of soil quality standards to ensure crop quality, animal and human health on this unique island chain.

This work aimed to investigate the metal accumulation characteristics as well as biogeochemical changes in the rhizosphere and root foraging strategies of this plant species. Previous reports suggested that Noccaea rotundifolia ssp. cepaeifolia is a Zn, Cd and Pb hyperaccumulator. We used hydroponic, rhizobox and split-pot experiments for studying metal accumulation and related rhizosphere processes. Although this species accumulated up to 1250 mg Pb kg−1 and 27,000 mg Zn kg−1 in shoots, translocation factors

Nickel (Ni) is used in numerous industrial processes, with large amounts of Ni-rich industrial wastes produced, which are largely sent to landfill. Nickel recovery from waste materials that would otherwise be disposed is of particular interest. Nickel phytomining represents a new technology in which hyperaccumulator plants are cultivated on Ni-rich substrates for commercial metal recovery. The aim of this study was to investigate the possibility of Ni transfer from industrial waste into plant biomass, to support recovery processes from bio-ores. Different industrial galvanic sludges (containing 85–150 g kg−1 Ni) were converted into artificial substrates (i.e. technosols) and the Ni hyperaccumulator Odontarrhena chalcidica (formerly Alyssum murale) was cultivated on these Ni-rich matrices. A greenhouse pot experiment was conducted for three months including an ultramafic soil control and testing fertilized (NPK) and unfertilized replicates. The results showed that fertilization was effective in improving plant biomass for all the substrates and that O. chalcidica was capable of viably growing on technosols, producing a comparable biomass to O. chalcidica on the control (ultramafic soil). On all technosols, O. chalcidica achieved Ni shoot concentrations of more than >1000 mg Ni kg −1 and maximum Ni uptake was obtained from one of the technosols (26.8 g kg −1 Ni, unfertilized; 20.2 g kg −1 Ni, fertilized). Nickel accumulation from three of the technosols resulted to be comparable with the control ultramafic soil. This study demonstrated the feasibility of transferring Ni from toxic waste into the biomass of Odontarrhena chalcidica and that phytomining from galvanic sludge-derived technosols can provide similar Ni yields as from natural ultramafic soils.

The remediation of petroleum hydrocarbon-contaminated soils has remained a challenging issue for environmental scientists and engineers. Within the scope of bioremediation; rhizoremediation has emerged as the most suitable method for petroleum-impacted soils. It can be promoted by the proper selection of suitable plant-microbe combinations and its overall efficiency can be enhanced by adding suitable soil amendments. This review article summarizes the available knowledge on rhizoremediation with respect to plant selection, inoculation with specific microbial strains (hydrocarbon-degrading as well as plant growth-promoting microbes) and a variety of soil amendments for enhancing the remediation efficiency. A large set of organic and inorganic soil amendments has been presented for the discussion on potential increase of rhizospheric effects. Finally, we discussed a unique multi-process phytoremediation system (MPPS) for the remediation of petroleum-polluted soils along with its field applications.

Co-contaminations of soils with organic and inorganic pollutants are a frequent environmental problem. Due to their toxicity and recalcitrance, the heterogeneous pollutants may persist in soil. The hypothesis of this study was that degradation of polycyclic aromatic hydrocarbons (PAHs) is enhanced if heavy metals in soil are immobilized and their bioavailability reduced. For metal immobilization and enhanced biodegradation, distinct mineral and organic soil amendments (iron oxides, gravel sludge, biochar) were deployed in an incubation batch experiment. The second part of the experiment consisted of a greenhouse pot experiment applying fast-growing and pollution-tolerant woody plants (willow and black locust). Soil amendments initially immobilized NH4NO3-extractable zinc, cadmium, and lead; after 100 days of incubation, soil amendments showed reductions only for cadmium and a tendency to enhance arsenic mobility. In order to monitor the remediation success, a 13C-phenanthrene (PHE) label was applied. 13C-phospholipid fatty acid analysis (13C-PLFA) further enabled the identification of PHE-degrading soil microorganisms. Both experiments exhibited a similar PLFA profile. Gram-negative bacteria (esp. cy17:0, 16:1ω7 + 6, 18:1ω7c) were the most significant microbial group taking up 13C-PHE. Plants effectively increased the label uptake by gram-positive bacteria and increased the biomass of the fungal biomarker, although their contribution to the degradation process was minor. Plants tended to prolong PAH dissipation in soil; at the end of the experiment, however, all treatments showed equally low total PAH concentrations in soil. While black locust plants tended not to take up potentially toxic trace elements, willows accumulated them in their leaves. The results of this study show that the chosen treatments did not enhance the remediation of the experimental soil.

The in situ stabilization of multielement-contaminated agricultural soils has limited effectiveness when using common single amendments. This study examined the use of drinking water treatment residues (WTR), based on (hydr)oxides of Fe, Al, or Mn, as a cost-effective solution to optimize the immobilization of metals (Cd, Pb, Zn) and As. Trace elements (TE) bioavailability was assessed under semi-controlled conditions in a pot study cultivating winter wheat (Triticum aestivum L. cv. Tiger) until maturity. An Fe-based WTR and a Mn-based WTR, applied at rates of 0.5 and 1% (m/m), were related to effects of lime marl (LM) application. Additionally, a bioassay with earthworms (Dendrobaena veneta) was conducted. Both bioassays were compared with measurements of NH4NO3-soluble, diffusive gradients in thin film (DGT)-available and soil solution TE concentrations, representing well-established surrogates for mimicking the bioavailable element fractions in soil. The application of the Fe-based WTR reduced As accumulation in vegetative wheat tissues (by up to 75%) and earthworms (by up to 41%), which corresponded with the findings from soil chemical analyses and improved plant growth and earthworm body weight. However, As concentrations in cereal grains were not affected, Cd or Pb accumulation by wheat was not mitigated, and Zn uptake was enhanced. By contrast, the Mn-based WTR effected the greatest reduction in Pb uptake, and lowered Cd transfer to wheat grain (by up to 25%). Neither the NH4NO3-soluble nor DGT-available concentrations matched with Cd and Zn accumulation in plants or earthworms, indicating interferences due to competition for binding sites according to the biotic ligand model. The results obtained in this study suggest that a bioassay with key species prior to field application should be mandatory when designing in situ stabilization options. The application of WTR to an agricultural soil strongly affected TE bioavailability to plants and earthworms. Low application rates tended to improve biomass production of biota. Higher application rates involved risks (e.g., P fixation, TE inputs), and none of the amendments tested could immobilize all targeted elements.

We investigated to which extent phytosiderophores (PS), released by grasses for the acquisition of iron, solubilize other metals in contaminated soils, and how this affects metal mobilization and uptake in wheat plants. A plant-based bioassay (‘RHIZOtest’) and batch extraction scheme were carried out for assessing metal mobilisation in soil, PS exudation and metal accumulation in wheat. Increased PS exudation was observed in Fe-deficient wheat, leading to enhanced Zn, Cu, Mn and Ni concentrations in wheat shoots on some soils. In contrast, plant Cd and Pb concentrations were not affected. Likewise, in the batch experiment, strongly increased extractable Cu, Ni and Zn concentrations were observed, in particular when 100 or 1000 μM PS were added. Our results suggest that Fe deficiency can enhance the accumulation of some metals in shoots of grass species. Although our results indicate that the risk of enhanced accumulation of Cd and Pb in Fe deficient wheat shoots is rather small, further experiments conducted on soil for the complete vegetation period would be needed to confirm this observation.

Phytomining could represent an innovative low-cost technology for the selective recovery of valuable trace elements from secondary resources. In this context the potential of phytomining from waste incineration bottom ash was tested in a pot experiment. Fresh bottom ash was acidified, leached to reduce salinity and amended with organic material to obtain a suitable substrate for plant growth. Two hyperaccumulator species, Alyssum serpyllifolium subsp. lusitanicum and Sedum plumbizincicola as well as three metal tolerant species, Brassica napus, B. juncea and Nicotiana tabacum were tested for their phytomining potential on the pre-treated and amended bottom ashes from municipal solid waste and hazardous waste incineration. The hyperaccumulators had severe difficulties to establish on the bottom ash and to produce sufficient biomass, likely due to salinity and Cu toxicity. Nevertheless, concentrations of Ni in A. serpyllifolium and Zn in S. plumbizincicola were high, but total metal removal was limited by the low biomass production and was clearly less than on metalliferous soils. The Brassica species proved to be more tolerant to salinity and high Cu concentrations and produced considerably higher biomass, but total metal removal was limited by rather low shoot concentrations. The observed limitations of the phytomining process along with currently low market prices of Ni and Zn suggest that further optimisation of the process is required in order to make phytomining economically feasible on the tested waste incineration bottom ashes.

International audience; Brownfield surface is expanding in Europe, but as often abandoned or underused, these areas become refuge for microbial, faunal and floral biodiversity. However, brownfield sites are generally contaminated, likely posing severe environmental risks. At a former wood preservation site contaminated with Cu, we evaluated the efficiency of compost and dolomitic limestone incorporation into the soil, followed by revegetation with Cu-tolerant grassy species, as a phytomanagement option to increase vegetation cover and plant diversity while reducing pollutant linkages. 7 years of phytomanagement enhanced natural revegetation through the improvement of soil physicochemical properties, particularly with compost-based amendments. The compost incorporation increased soil Cu solubility; however, no increment in Cu availability and a reduction in Cu-induced phytotoxicity were observed with the compost. The improved soil nutrient availability and the soil phytotoxicity mitigation in compost-amended soils facilitated over the 7 years the growth of beneficial plant colonists, including leguminous species, which can potentially promote essential soil functions. Soil treatments did not affect Cu uptake and translocation by plants and shoot Cu levels indicated no risk for the food chain. Overall, a long-term phytomanagement combining an initial amendment of compost and dolomitic limestone with the cultivation of Cu-tolerant grassy populations can ameliorate such Cu-contaminated soils, by mitigating risks induced by Cu excess, ultimately allowing the development of a meadow that can provide ecological and economic benefits in terms of ecosystem services.Agrostis capillaris; Agrostis gigantea; Metal pollution; Organic amendments; Phytotoxicity; Phytoremediation

International audience; Ultramafic soils are typically enriched in nickel (Ni), chromium (Cr), and cobalt (Co) and deficient in essential nutrients, making them unattractive for traditional agriculture. Implementing agromining systems in ultramafic agricultural soils represent an ecological option for the sustainable management and re-valorisation of these low-productivity landscapes. These novel agroecosystems cultivate Ni-hyperaccumulating plants which are able to bioaccumulate this metal in their aerial plant parts; harvested biomass can be incinerated to produce Ni-enriched ash or “bio-ore” from which Ni metal, Ni ecocatalysts or pure Ni salts can be recovered. Nickel hyperaccumulation has been documented in ∼450 species, and in temperate latitudes these mainly belong to the family Brassicaceae and particularly to the genus Odontarrhena (syn. Alyssum pro parte). Agromining allows for sustainable metal recovery without causing the environmental impacts associated with conventional mining activities, and at the same time, can improve soil fertility and quality and provide essential ecosystem services. Parallel reductions in Ni phytotoxicity over time would also permit cultivation of conventional agricultural crops. Field studies in Europe have been restricted to Mediterranean areas and these only evaluated the Ni-hyperaccumulator Odontarrhena muralis s.l. Two recent EU projects (Agronickel and LIFE-Agromine) have established a network of agromining field sites in ultramafic regions with different edapho-climatic characteristics across Albania, Austria, Greece and Spain. Soil and crop management practices are being developed so as to Kidd et al. Sustainable Agromining Systems for Nickel Recovery optimize the Ni agromining process; field studies are evaluating the potential benefits of fertilization regimes, crop selection and cropping patterns, and bioaugmentation with plant-associated microorganisms. Hydrometallurgical processes are being up-scaled to produce nickel compounds and energy from hyperaccumulator biomass. Exploratory techno-economic assessment of Ni metal recovery by pyrometallurgical conversion of O. muralis s.l. shows promising results under the condition that heat released during incineration can be valorized in the vicinity of the processing facility.

The pollutant input and the risk of contamination by long-term application of recycled P fertilizers (RPFs) in European agricultural soils were estimated by a mass balance approach. Calculations based on literature data were carried out for the potentially toxic elements (PTEs) Cd, Cr, Cu, Ni, Pb, Zn and for the persistent organic pollutants (POPs) PCBs, PAHs and polychlorinated dibenzo-dioxins and -furans. The PTE accumulation estimation during 200 years of fertilizer application, equivalent to 11 kg P ha−1 year−1, the mean P export via harvested products on European stockless organic farms, regarded soil background concentrations and proposed threshold concentrations. For PTEs, inputs were fertilizer application, atmospheric deposition and liming, output processes were leaching and crop harvest. The effect of varying site conditions was assessed by considering two precipitation excess (F) values and two soil pH values. For POPs, fertilizer application and the half-life time were considered. The PTE accumulation risk was low for most RPFs. For the analysed POPs no accumulation was found. The highest accumulation was found for all PTEs at pH 7 and F = 0.1 m year−1, the lowest at pH 5 and F = 0.3 m year−1. A high P concentration in fertilizers resulted in a low PTE flow per unit of P. Composts had the highest PTE accumulation risk due to lowest P contents. Struvite, meat and bone meal, digestate of catering waste, ash and biosolids would be better suited as P fertilizers. The use of fertilizers should be regulated based on their pollutant-to-nutrient ratio.

Gentle remediation options (GRO), i.e. in situ stabilisation, (aided) phytoextraction and (aided) phytostabilisation, were implemented at ten European sites contaminated with trace elements (TE) from various anthropogenic sources: mining, atmospheric fallout, landfill leachates, wood preservatives, dredged-sediments, and dumped wastes. To assess the performance of the GRO options, topsoil was collected from each field trial, potted, and cultivated with lettuce (Lactuca sativa L.) for 48 days. Shoot dry weight (DW) yield, photosynthesis efficiency and major element and TE concentrations in the soil pore water and lettuce shoots were measured.GRO implementation had a limited effect on TE concentrations in the soil pore water, although use of multivariate Co-inertia Analysis revealed a clear amelioration effect in phytomanaged soils. Phytomanagement increased shoot DW yield at all industrial and mine sites, whereas in agricultural soils improvements were produced in one out of five sites. Photosynthesis efficiency was less sensitive than changes in shoot biomass and did not discriminate changes in soil conditions.Based on lettuce shoot DW yield, compost amendment followed by phytoextraction yielded better results than phytostabilisation; moreover shoot ionome data proved that, depending on initial soil conditions, recurrent compost application may be required to maintain crop production with common shoot nutrient concentrations.

Characterisation of geochemical transformations and processes in soils with special focus on the rhizosphere is crucial for assessing metal(loid) bioavailability to plants during in situ immobilisation and phytostabilisation. In this study, the effects of nano zero-valent iron (nZVI) were investigated in terms of the immobilisation of As, Zn, Pb and Cd in two soil types and their potential uptake by plants using rhizobox experiments. Such system allowed monitoring the behaviour of trace elements in rooted and bulk soil compartments separately. Sunflower (Helianthus annuus L.) and ryegrass (Lolium perenne L.) were tested for As-rich (15.9 g As kg−1) and Zn-rich (4.1 g Zn kg−1) soil samples, respectively. The application of nZVI effectively lowered the uptake of all target risk elements into plant tissues. Efficient immobilisation of As was determined in the As-soil without a significant difference between plant and bulk soil compartments. Similarly, a significant decrease was determined for CaCl2-available fractions of Zn, Pb and Cd in nZVI-treated Zn-soil. The behaviour of As corresponded to changes in Eh, while Zn and Cd showed to be mainly pH-dependent. However, despite the observed stabilisation effect of nZVI, high amounts of As and Zn still remained available for plants. Furthermore, the accumulation of the target risk elements in roots and the overall effect of nZVI transformations in the rhizosphere were verified and visualised by SEM/EDS. The following immobilising mechanisms were suggested: (i) sorption onto both existing and newly formed Fe (hydr)oxides, (ii) formation of secondary Fe-As phases, and (iii) sorption onto Mn (hydr)oxides.

In soil, mixed contamination with potentially toxic trace elements and polycyclic aromatic hydrocarbons (PAHs) may persist for a long time due to strong adsorption to the soil matrix and to its toxicity to microorganism. We conducted an incubation batch experiment to test the effect of soil amendments (biochar, gravel sludge, iron oxides) on the immobilisation of trace elements. To monitor microbial degradation, a 13C-PHE (phenanthrene) label was introduced to soil for 13C-PLFA (phospholipid fatty acid) analysis. Soil amendments increased soil pH, reduced mobility of NH4NO3-extractable trace elements Cd and Zn, and increased mobile Cu. A small consortium of PHE degraders was identified mainly in the microbial groups of gram-negative bacteria and actinomycetes. The degradation process of PHE peaked 9days after incubation start. PAH concentrations remained constant in the soil within the 30-day incubation, except for the easily available 13C-PHE in the amended treatment. In order to test the effect of plants and soil amendments under more realistic conditions, we also conducted an outdoor pot experiment with black locust (Robinia pseudoacacia Nyirsegi). Furthermore, soil amendments increased the mobility of soil Cu and As and decreased the mobility of Cd, Pb and Sb. The uptake of trace elements to leaves was low. Σ 16 U.S. EPA PAHs were significantly reduced only in the combined treatment of black locust and soil amendments after 12months of plant growth. Soil amendment-assisted phytoremediation showed a high efficiency in PAH dissipation and may be a useful remediation technique for mixed contaminated soils.

Environmental problems such as the deterioration of groundwater quality, soil degradation and various threats to human, animal and ecosystem health are closely related to the presence of high concentrations of organic xenobiotics in the environment. Employing appropriate technologies to remediate contaminated soils is crucial due to the site-specificity of most remediation methods. The limitations of conventional remediation technologies include poor environmental compatibility, high cost of implementation and poor public acceptability. This raises the call to employ biological methods for remediation. Bioremediation and microbe-assisted bioremediation (phytoremediation) offer many ecological and cost-associated benefits. The overall efficiency and performance of bio- and phytoremediation approaches can be enhanced by genetically modified microbes and plants. Moreover, phytoremediation can also be stimulated by suitable plant-microbe partnerships, i.e. plant-endophytic or plant-rhizospheric associations. Synergistic interactions between recombinant bacteria and genetically modified plants can further enhance the restoration of environments impacted by organic pollutants. Nevertheless, releasing genetically modified microbes and plants into the environment does pose potential risks. These can be minimized by adopting environmental biotechnological techniques and guidelines provided by environmental protection agencies and other regulatory frameworks. The current contribution provides a comprehensive overview on enhanced bioremediation and phytoremediation approaches using transgenic plants and microbes. It also sheds light on the mitigation of associated environmental risks.

Besides carbon sequestration and improvement of soil properties, biochar (BC) has increasingly been studied as an amendment to immobilise heavy metals in contaminated soils. In a 2-year experiment, we analysed the effects of poplar BC (P-BC, mixed with compost) and gravel sludge with siderite-bearing material (GSFe) on a Cd-, Pb- and Zn-contaminated soil and on metal concentration in Miscanthus × giganteus shoots under greenhouse and field conditions. In the greenhouse, 1% (m/m) P-BC addition reduced NH4NO3-extractable Cd, Pb and Zn concentrations by 75, 86 and 92%, respectively, at the end of the study. In the leachates, P-BC (1%) could significantly reduce Cd and Zn in both years. In the field, P-BC (3%) induced a reduction of extractable Cd by 87% whereas a combination of P-BC + GSFe reduced Pb by 82% and Zn by 98% in the first year and by 83 and 96% in the second year. In contrast, the metal immobilisation in the soil was hardly reflected in the shoots of Miscanthus × giganteus which generally showed metal concentrations close to control. While Cd was not influenced in both years, Pb and Zn were slightly reduced. Our study confirmed that Miscanthus is an efficient metal excluder, corroborating its suitability for the production of renewable biomass on metal-contaminated soils.

Waste incineration bottom ash is considered a secondary resource for valuable trace elements (TE), which is currently neglected in most European countries. Phytomining could potentially recover valuable TE from such waste materials but is still at an exploratory stage with many challenges. The use of bioaugmentation to improve plant growth and TE accumulation of metal-tolerant high biomass plants growing on waste incineration bottom ash was evaluated. Bacterial strains that were previously isolated from rhizosphere, roots and contaminated soil were selected according to their plant growth promoting characteristics and tolerance to the bottom ash substrate. Those selected bacterial strains were tested for their beneficial effects on Nicotiana tabacum and Salix smithiana with regards to phytomining. The rhizobacterial strain Rhodococcus erythropolis P30 enhanced the shoot dry weight of N. tabacum by on average 57% compared to the control plants. Several bacterial inoculants enhanced biomass production and the nutritional status of S. smithiana. Moreover, those bacterial strains previously described to enhance biomass production of N. tabacum and members of the Salicaceae on TE-contaminated soils, also enhanced biomass production of these species on bottom ash. However, bacterial inoculants could not enhance trace element accumulation in plants.

11 páginas.-- 4 figuras.-- 6 tablas.-- 40 referencias, Purpose: This study evaluates the effects of two soil amendments and the growth of two plant species on labile trace element (TE) fractions in two different contaminated soils. Materials and methods: We studied the effects of two organic amendments (biosolid compost and alperujo compost) and two plant species (Medicago polymorpha and Poa annua) on pH, total organic carbon (TOC), and TE availability, by three extraction methods (CaCl2 aqueous solution, soil pore water (SPW), and diffusive gradient in thin film (DGT)), in two contaminated soils with contrasting pH values (Aznalcázar, 6.53, and Vicario, 3.48) in a 118-day pot experiment. The effects of the composts on labile TE fractions were compared with element concentrations in plants. Results and discussion: No relevant effects of amendments and plants were found on the physical and chemical characteristics of the Aznalcázar soil. However, the addition of amendments was essential for plant species growing in the acid Vicario soil. In this soil, amendments and plant growth increased pH and TOC and reduced substantially TE bioavailability. Although absolute values of bioavailable TE contents obtained by the three methods were very different and followed the trend CaCl2 extraction > SPW > DGT, these values follow a similar behavior in the two studied soils and for the two species. Conclusions: The results demonstrate that the application of organic amendments are suitable for remediating acid TE-contaminated soils, for the establishment of a vegetation cover on previously bare soils for reducing wind and water erosion and for reducing labile TE fractions to prevent leaching of pollutants into subsoil or groundwater layers. Moreover, the results obtained in this study pointed out that under microcosm conditions, the three methods tested (CaCl2 extraction, SPW, and DGT) to predict TE bioavailability were highly correlated. © 2015 Springer-Verlag Berlin Heidelberg, AGL2011-23617 was supported by the CICYT of the Ministerio de Ciencia e Innovación of Spain and FEDER (EU). L. Ciadamidaro thanks the CSIC for funding her grant (JAE-PreDoc).

A questionnaire survey was carried out in four European countries to gather end-user's perceptions of using plants from phytotechnologies in combustion and anaerobic digestion (AD). Nine actors of the wood energy sector from France, Germany, and Sweden, and eleven AD platform operators from France, Germany, and Austria were interviewed. Questions related to installation, input materials, performed analyses, phytostabilization, and phytoextraction were asked. Although the majority of respondents did not know phytotechnologies, results suggested that plant biomass from phytomanaged areas could be used in AD and combustion, under certain conditions. As a potential benefit, phytomanaged plants would not compete with plants grown on agricultural lands, contaminated lands being not suitable for agriculture production. Main limitations would be related to additional controls in process' inputs and end-products and installations that might generate additional costs. In most cases, the price of phytotechnologies biomass was mentioned as a driver to potentially use plants from metal-contaminated soils. Plants used in phytostabilization or phytoexclusion were thought to be less risky and, consequently, benefited from a better theoretical acceptance than those issued from phytoextraction. Results were discussed according to national regulations. One issue was related to the regulatory gap concerning the status of the plant biomass produced on contaminated land.

Arsenic and Cu mobility was investigated in the rhizosphere of Lupinus albus L. grown in an iron-amended contaminated soil. White lupin was grown in rhizobags in contaminated soil either left untreated or amended with iron sulphate plus lime (Fe + lime) or biochar (Fe + BC). Porewater was monitored in rhizosphere and bulk soil throughout the experiment and the extractable fraction of several elements and As and Cu plant uptake was analysed after 48 days. The distribution of As, Cu, P and Fe in the lupin rhizosphere was evaluated with chemical images obtained by laser ablation-ICP-MS analysis of diffusive gradients in thin films (DGT) gels. The treatments effectively reduced the soluble and extractable As and Cu fractions in the bulk soil, but they did not affect plant uptake. In all cases, soluble As was slightly enhanced in the rhizosphere. This difference was more pronounced in the Fe + lime-treated rhizosphere soil, where an increase of pH as well as extractable As and Fe concentrations were also observed. Chemical imaging of the lupin rhizosphere also showed slightly higher As- and Fe-DGT fluxes around lupin roots grown in the non-amended soil. Our findings indicate As and Fe co-solubilisation by lupin root exudates, likely as a response to P deficiency. Arsenic mobilisation occurred only in the rhizosphere and was not decreased by the amendments.

Trace element-contaminated soils (TECSs) are one of the consequences of the past industrial development worldwide. Excessive exposure to trace elements (TEs) represents a permanent threat to ecosystems and humans worldwide owing to the capacity of metal(loid)s to cross the cell membranes of living organisms and of human epithelia, and their interference with cell metabolism. Quantification of TE bioavailability in soils is complicated due to the polyphasic and reactive nature of soil constituents. To unravel critical factors controlling soil TE bioavailability and to quantify the ecological toxicity of TECSs, TEs are pivotal for evaluating excessive exposure or deficiencies and controlling the ecological risks. While current knowledge on TE bioavailability and related cumulative consequences is growing, the lack of an integrated use of this concept still hinders its utilization for a more holistic view of ecosystem vulnerability and risks for human health. Bioavailability is not generally included in models for decision making in the appraisal of TECS remediation options. In this review we describe the methods for determining the TE bioavailability and technological developments, gaps in current knowledge, and research needed to better understand how TE bioavailability can be controlled by sustainable TECS management altering key chemical properties, which would allow policy decisions for environmental protection and risk management.

Sunflower plants were grown on soil contaminated with several levels of Cd (from 0.3 to 15 mg Cd kg/soil). Cd and nutrient (Cu, Zn, Fe, Ca, K, Mg) contents were evaluated in root and aboveground biomass during the vegetative growth period (from the emergence of the cotyledon leaves to the flower bud phase). An overall increase of Cd content was found in the plants during the growing cycle. Significant interactions were found between Cd contamination levels and phenological stages for Cu, Fe, Ca and Mg content in the whole plant. Cd levels involved significant increases of such nutrients only at the flower bud phase. At this stage, significant interactions between Cd levels in soil and plant portions (root and shoot) were found only for micronutrients (Zn, Cu, Fe). The translocation of micronutrients from root to the shoot was negatively affected by Cd, which probably interferes with the mechanisms of the element transport in the plant. Root stored about 70% of the total Cd and showed greater imbalance in nutrient content compared to the aboveground portion. Cadmium affects plant nutritive status in a different way depending on Cd contamination level, nutrient considered, plant portion and phenological stage.

Gentle remediation options (GRO) are based on the combined use of plants, associated microorganisms and soil amendments, which can potentially restore soil functions and quality. We studied the effects of three GRO (aided-phytostabilisation, in situ stabilisation and phytoexclusion, and aided-phytoextraction) on the soil microbial biomass and respiration, the activities of hydrolase enzymes involved in the biogeochemical cycles of C, N, P, and S, and bacterial community structure of trace element contaminated soils (TECS) from six field trials across Europe. Community structure was studied using denaturing gradient gel electrophoresis (DGGE) fingerprinting of Bacteria, α - and β - Proteobacteria , Actinobacteria and Streptomycetaceae , and sequencing of DGGE bands characteristic of specific treatments. The number of copies of genes involved in ammonia oxidation and denitrification were determined by qPCR. Phytomanagement increased soil microbial biomass at three sites and respiration at the Biogeco site (France). Enzyme activities were consistently higher in treated soils compared to untreated soils at the Biogeco site. At this site, microbial biomass increased from 696 to 2352 mg ATP kg −1 soil, respiration increased from 7.4 to 40.1 mg C-CO 2 kg −1 soil d −1 , and enzyme activities were 2–11-fold higher in treated soils compared to untreated soil. Phytomanagement induced shifts in the bacterial community structure at both, the total community and functional group levels, and generally increased the number of copies of genes involved in the N cycle ( nirK , nirS , nosZ , and amoA ). The influence of the main soil physico-chemical properties and trace element availability were assessed and eventual site-specific effects elucidated. Overall, our results demonstrate that phytomanagement of TECS influences soil biological activity in the long term.

Human activities lead to increasing concentration of the stable elements cesium (Cs) and strontium (Sr) and their radioactive isotopes in the food chain, where plants play an important part. Here we investigated Plantago major under the influence of long-term exposure to stable Cs and Sr. The plants were cultivated hydroponically in different concentrations of cesium sulfate (between 0.002 and 20 mM) and strontium nitrate (between 0.001 and 100 mM). Uptake of Cs and Sr into leaves was analyzed from extracts by inductively coupled plasma mass spectrometry (ICP-MS). It was increased with increasing external Cs and Sr concentrations. However, the efficiency of Cs and Sr transfer from solution to plants was higher for low external concentrations. Highest transfer factors were 6.78 for Cs and 71.13 for Sr. Accumulation of Sr was accompanied by a slight decrease of potassium (K) and calcium (Ca) in leaves, whereas the presence of Cs in the medium affected only uptake of K. The toxic effects of Cs and Sr were estimated from photosynthetic reactions and plant growth. In leaves, Cs and Sr affected the chlorophyll fluorescence even at their low concentrations. Low and high concentrations of both ions reduced dry weight and length of roots and leaves. The distribution of the elements between the different tissues of leaves and roots was investigated using Energy Dispersive X-Ray microanalysis (EDX) with scanning electron microscope (SEM). Overall, observations suggested differential patterns in accumulating Cs and Sr within the roots and leaves. When present in higher concentrations the amount of Cs and Sr transferred from environment to plants was sufficient to affect some physiological processes. The experimental model showed a potential for P. major to study the influence of radioactive contaminants and their removal from hotspots.

Phytomanagement of metal-polluted soils requires information on plant responses to metal availability in soil, but the predictability of metal accumulation in plant shoots and/or roots may be limited by metal toxicity and inherent shortfalls of the bioavailability assays. We measured the uptake of Cd and Zn in a Salix smithiana clone grown in a pot experiment on soils with different characteristics and metal availabilities, determined by conventional soil single extractions (0.05 M Na2-EDTA and 1 M NH4NO3), soil solution obtained by centrifugation, and diffusive gradients in thin films (DGT). The Cd and Zn phytoavailability after a 2-year phytoextraction by willow was assessed by metal accumulation in the straw of the following barley culture. The phytoextraction efficiency was largest on a moderately polluted acid soil. Biomass and shoot Zn concentrations of S. smithiana were better predicted by DGT-measured Zn concentrations in soil solution (C DGT) than by Zn concentrations in the soil solution and extractable soil fractions. The weaker correlation for Cd in shoots may be related to relative Cd enrichment in the plant tissues. The metal accumulation in barley straw was unaffected or increased after a 2-year phytoextraction. The shoot Zn and Cd removal of the tested Salix clone can be predicted by C DGT concentrations and is highest on either calcareous or moderately polluted acid soils. Single extraction with NH4NO3 and the C DGT value of Cd were not able to predict shoot Cd removal on the tested soils. Only shoot removal of Zn was predicted fairly well by the C DGT value.

Arsenic is a non-threshold carcinogenic metalloid. Thus, human exposure should be minimised, e.g. by chemically stabilizing As in soil. Since iron is a potential As immobiliser, it was investigated whether root iron plaque, formed under aerobic conditions, affects As uptake, metabolism and distribution in Lupinus albus plants. White lupin plants were cultivated in a continuously aerated hydroponic culture containing Fe/EDDHA or FeSO4 and exposed to arsenate (5 or 20 μM). Only FeSO4 induced surficial iron plaque in roots. LA-ICP-MS analysis accomplished on root sections corroborated the association of As to this surficial Fe. Additionally, As(V) was the predominant species in FeSO4-treated roots, suggesting less efficient As uptake in the presence of iron plaque. Fe/EDDHA-exposed roots neither showed such surficial FeAs co-localisation nor As(V) accumulation; in contrast As(III) was the predominant species in root tissue. Furthermore, FeSO4-treated plants showed reduced shoot-to-root As ratios, which were >10-fold lower compared to Fe/EDDHA treatment. Our results highlight the role of an iron plaque formed in roots of white lupin under aerobic conditions on As immobilisation. These findings, to our knowledge, have not been addressed before for this plant and have potential implications on soil remediation (phytostabilisation) and food security (minimising As in crops).

Gentle remediation options (GROs) are risk management strategies or technologies involving plant (phyto-), fungi (myco-), and/or bacteria-based methods that result in a net gain (or at least no gross reduction) in soil function as well as effective risk management. GRO strategies can be customised along contaminant linkages, and can generate a range of wider economic, environmental and societal benefits in contaminated land management (and in brownfields management more widely). The application of GROs as practical on-site remedial solutions is still limited however, particularly in Europe and at trace element (typically metal and metalloid) contaminated sites. This paper discusses challenges to the practical adoption of GROs in contaminated land management, and outlines the decision support tools and best practice guidance developed in the European Commission FP7-funded GREENLAND project aimed at overcoming these challenges. The GREENLAND guidance promotes a refocus from phytoremediation to wider GROs- or phyto-management based approaches which place realisation of wider benefits at the core of site design, and where gentle remediation technologies can be applied as part of integrated, mixed, site risk management solutions or as part of “holding strategies” for vacant sites. The combination of GROs with renewables, both in terms of biomass generation but also with green technologies such as wind and solar power, can provide a range of economic and other benefits and can potentially support the return of low-level contaminated sites to productive usage, while combining GROs with urban design and landscape architecture, and integrating GRO strategies with sustainable urban drainage systems and community gardens/parkland (particularly for health and leisure benefits), has large potential for triggering GRO application and in realising wider benefits in urban and suburban systems. Quantifying these wider benefits and value (above standard economic returns) will be important in leveraging funding for GRO application and soft site end-use more widely at vacant or underutilized sites.

High-biomass tree species holds promise for a clean up of metal contaminated soils. Root and fungal activities modify soil characteristics that are important factors for the phytoextraction process (metal availability and toxicity). In a rhizobox experiment, two clones of Salix × rubens derived from contaminated and non-contaminated sites were tested for growth performance and metal (Cd, Pb and Zn) accumulation on a polluted Calcaric Cambisol. The largest metal concentrations in leaves were 66.7 mg Cd/kg, 12.8 mg Pb/kg and 1090 mg Zn/kg. The results indicate that metal tolerance and accumulation of S. × rubens may be a constitutive rather than an adaptive property. Soil pH did not differ among rhizobox compartments. However, acid neutralization capacity was decreased in rhizosphere. DOC in rhizosphere was increased by 37% and seemed to enhance labile fraction of Pb and Zn, whereas Cd was not affected. The replenishment of labile metals from less labile soil fractions was efficient enough to almost compensate the plant uptake. S. × rubens can effectively induce chemical changes in the rhizosphere is very promising for a clean up of metal-polluted soils.

Heavy metal contamination affects large areas of Europe and worldwide. Hot spots of pollution are located close to industrial sites, around large cities and in the vicinity of mining and smelting plants. Agriculture in these areas faces major problems due to heavy metal transfer into crops and subsequently into the food chain. This paper gives an overview on simple but effective countermeasures to reduce the transfer of heavy metals to edible parts of crops. Since crop species and varieties largely differ in their heavy metal uptake, choosing plants with low transfer factors (e.g., legumes, cereals) may reduce metal concentration in edible parts significantly. Cultivating crops with higher

During the past decades a number of field trials with gentle remediation options (GRO) have been established on trace elements (TE) contaminated sites throughout Europe. Each research group selects different methods to assess the remediation success making it difficult to compare efficacy between various sites and treatments. This study aimed at selecting a minimum risk assessment battery combining chemical and ecotoxicological assays for assessing and comparing the effectiveness of GRO implemented in seven European case studies. Two test batteries were pre-selected; a chemical one for quantifying TE exposure in untreated soils and GRO-managed soils and a biological one for characterizing soil functionality and ecotoxicity. Soil samples from field studies representing one of the main GROs (phytoextraction in Belgium, Sweden, Germany and Switzerland, aided phytoextraction in France, and aided phytostabilisation or in situ stabilization/phytoexclusion in Poland, France and Austria) were collected and assessed using the selected test batteries. Extractable TE concentrations generally decreased more significantly in soils managed by in situ stabilisation combined with phytoexclusion, phytostabilisation or phytoextraction than in soils only managed with phytoextraction. Pseudo-total TE concentrations did not change in the phytomanaged sites, except for one case, which was attributed to the dilution by the amendments. Among the single chemical extractions, the NH4NO3- and EDTA-extractions showed most frequently the differences in the extracted TE concentrations between the treated and untreated soils, while the most frequent correlations with the biological responses occurred for NH4NO3, followed by NaNO3-extractable TE. Pseudo-total (aqua regia extractable) concentrations showed weak correlation with the biological responses. Out of the bioindicators (plants, earthworms, and nematodes), dwarf beans, especially through root mass, followed by shoot length, and stress enzyme activities, were the most responsive indicators to the soil treatments. Even though the selective chemical extractions did not always show statistically significant changes in TE extractability, dwarf beans and stress enzymes developed a stronger response to the tested GRO options. Generally, the plant growth decreased with higher extractable TE concentrations in soil, while bean stress enzymes reacted in the opposite way, i.e. increased with increasing TE extractability. It is suggested that a minimum risk assessment battery to compare or monitor the sites phytomanaged by GROs might consist of the NH4NO3-extraction and the dwarf bean Plantox test including the stress enzyme activities. Validation of this assessment battery as a potential indicator of successful remediation should be further implemented for a broader range of soils and long-term treatments.

International audience; Gentle Remediation Options (GRO) are risk management strategies or techniques for contaminated sites that result in no gross reduction in soil functionality (or a net gain) as well as risk management. Intelligently applied GROs can provide: (a) rapid risk management via pathway control, through containment and stabilisation, coupled with a longer term removal or immobilisation/isolation of the contaminant source term; and (b) a range of additional economic (e.g. biomass generation), social (e.g. leisure and recreation) and environmental (e.g. CO2 sequestration) benefits. In order for these benefits to be optimised or indeed realised, effective stakeholder engagement is required. This paper reviews current sector practice in stakeholder engagement and its importance when implementing GRO and other remediation options. From this, knowledge gaps are identified, and strategies to promote more effective stakeholder engagement during GRO application are outlined. Further work is required on integrating stakeholder engagement strategies into decision support systems and tools for GRO (to raise the profile of the benefits of effective stakeholder engagement and participation, particularly with sector professionals), and developing criteria for the identification of different stakeholder profiles/categories. Demonstrator sites can make a significant contribution to stakeholder engagement via providing evidence on the effectiveness of GRO under varying site contexts and conditions. Effective and sustained engagement strategies however will be required to ensure that site risk is effectively managed over the longer-term, and that full potential benefits of GRO (e.g. CO2 sequestration, economic returns from biomass generation and “leverage” of marginal land, amenity and educational value, ecosystem services) are realised and communicated to stakeholders.

The use of fast growing trees is a common practice for phytoremediation of contaminated soils. Plant roots can change trace element bioavailability in soils. We studied the effect of Populus alba on trace element bioavailability on two contaminated soils (one with neutral pH and other with acid pH) comparing two methods (0.01M CaCl2-extractable in soil and concentration in soil pore water SPW), trace element accumulation in leaves and plant development over 36months. Results were compared to those obtained with a non-contaminated soil. The experiment was carried out in containers (95L of volume and 1m height). Half of the containers for each soil were planted with P. alba saplings and the others remained without plant. In neutral soils plant growth did not influence soil pH; the greatest effect due to plant growth was found in acid soil. Values of pH obtained by SPW showed a similar trend compared to those obtained after soil KCl extraction. Bioavailability of trace elements determined by both methods followed the same behavior in the three studied soils. Both methods for determining trace element bioavailability in soil were accurate to predict plant uptake. In non-contaminated soil, plants tended to increase micronutrients (Cu, Mn and Zn) availability. However, in case of contaminated soil, the growth of P. alba did not increase trace element availability. Moreover, results on height and diameter of the trunk of the trees, during 36months, demonstrated that the presence of total trace elements in soil did not affect plant development., AGL2008-00985 is a support from the CICYT of the Ministerio de Ciencia e Innovación of Spain and the FEDER (EU). L. Ciadamidaro thanks CSIC for funding his grant (JAE-PreDoc).

Complément de : "European Union ERANET SNOWMAN project SUMATECS (Sustainable Management of Trace Element Contaminated Sites)"; International audience; A range of tools have been proposed to support decision making in contaminated land remediation. From a European perspective it is clear, however, that there are considerable national differences in the decision support process, and more generally in the extent to which this process supports the selection of less invasive, alternative remediation options such as phytoremediation, in situ immobilisation etc. (referred to here as “gentle” remediation technologies). In this paper we present results from the recently completed European Union ERANET SNOWMAN project SUMATECS (Sustainable Management of Trace Element Contaminated Sites), and critically review available decision support tools in terms of their fitness for purpose for the application of gentle remediation technologies. Stakeholder feedback indicates a lack of knowledge amongst stakeholders of currently available decision support tools. We propose that decision support which focuses on gentle remediation is more strongly incorporated into existing, well-established (national) decision support tools / decision-frameworks, to promote more widespread use and uptake.

The understanding of the influence of toxic elements on root anatomy and element distribution is still limited. This study describes anatomical responses, metal accumulation and element distribution of rooted cuttings of Salix caprea after exposure to Cd and/or Zn. Differences in the development of apoplastic barriers and tissue organization in roots between two distinct S. caprea isolates with divergent Cd uptake and accumulation capacities in leaves might reflect an adaptive predisposition based on different natural origins. Energy-dispersive X-ray spectroscopy (EDX) revealed that Cd and Zn interfered with the distribution of elements in a tissue- and isolate-specific manner. Zinc, Ca, Mg, Na and Si were enriched in the peripheral bark, K and S in the phloem and Cd in both vascular tissues. Si levels were lower in the superior Cd translocator. Since the cuttings originated from stocks isolated from polluted and unpolluted sites we probably uncovered different strategies against toxic elements., Highlights ► We describe responses in roots of S. caprea exposed to Cd and Zn. ► Apoplastic barrier development varied among isolates from differently polluted sites. ► EDX analyses revealed variations of element distributions in root tissues. ► Si weight% was lower in the isolate with a higher Cd translocation capacity. ► S. caprea isolates possessed different strategies to respond to Cd and Zn., S. caprea altered element distribution and translocation, apoplastic barrier development and root anatomy upon Cd and/or Zn exposure.

The effects of soil cadmium (Cd) contamination on Cd accumulation and distribution, growth and physiological responses of sunflower plants were investigated. Plants were subject to six levels of soil contamination (from 2.5 to 15 mg Cd kg/soil) with an untreated control, from the emergence of the cotyledon leaves until the harvest, when plants were at the flower bud stage. An overall increase of Cd concentration was found in all tissues of the plants (roots, stem, young, mature and old leaves) by increasing the Cd contamination in the soil. Regardless of treatments, Cd concentration in roots always exceeded those in the aboveground dry matter with a low translocation from roots to shoots. At early stage of growth, Cd concentration in plants was higher than at the flower bud stage. Soil Cd contamination did not affect plant growth, relative water content and gas exchange parameters. Negative and significant correlation was only found between Cd concentration in the young leaves and chlorophyll concen tration at the end of vegetative growing stage. Roots and old leaves are the main metal sinks suggesting a defense or tolerance mechanism of the plants to avoid toxic levels in physiologically most active apical tissues. These results should be tested in open field to verify the suitability of sunflower in the area of phytotechnologies.

We conducted two laboratory experiments to examine the effectsoffertilisationandagitation(aeration)oncrudeoil degradation in two soils with differentialnutrient(nitrogen,phosphorus)availability.Twosoilsthathadbeenspiked with crude oil two years before were mixed with nitrogen and/or phosphorus at three differentlevelsandsubsequently incubated 28 days (Exp. 1). In experiment 2 we investigated the effectofrepeatedagitation(manualmixing) on hydrocarbon degradation with and without fertilisation. One of the soils was also freshly spiked to as-sess the impact of ageing. Heptane-extractable hydrocarbon concentrations were determined in both experiments and substrate-induced respiration in Exp. 2. Thegenerallysmallchangesofhydrocarbonconcentrationsduring28days of incubation in Exp. 1 are likely attributed to low bio-accessibility of hydrocarbons as a consequence of long-term ageing. Fertilization of nitrogen, phosphorus or combinations thereof was ineffectiveinmosttreatmentsofExp. 1, which may be explained by limited oxygen supply due to the high proportions of clay and silt. However, agi-tation enhanced HC biodegradation in the sandy-loamy soil by about 15% (Exp. 2) compared to non-agitated treat-ments. In contrast, we observed no effectofagitationinthesandysoil.Keywords: biodegradation; crude oil; hydrocarbon; fertiliser; contaminated soils; incubation