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Purpose The present opinion paper is focused on the phytoremediation of organic pollutants and is based on the lectures given by the author during the International Congress of Phytoremediation of Polluted Soils, held at Vigo, Spain, 29–30 July 2014. The aim of this position paper is (1) to highlight some progress made within the last few years (after the end of COST Action 859 in October 2009) in the phytoremediation of selected organic pollutants and (2) to suggest new research and approaches, which seem important and promising in the opinion of the author to make this environmentally friendly remediation technique more attractive and more successful. Results and discussion Depending on the type of soil to be treated, as well as on the xenobiotic contaminants, their concentration and ageing, different approaches can be considered and are briefly presented with some recent and successful applications, but also highlighting their limitations and needs for future developments: phytoextraction of hydrophobic xenobiotic compounds like polychlorobiphenyls (PCBs), phytodegradation of xenobiotics and its possible impacts on primary and secondary metabolism of the plant and phytostimulation of rhizospheric microorganisms by root exudates for the rhizodegradation of petroleum hydrocarbons. Outlook Finally, some promising approaches are suggested for overcoming the bottlenecks and making phytoremediation a reliable, mature and sustainable technology: how to deal with mixed pollution; the potential of endophytic bacteria; possible improvements by soil amendments and co-cropping; validation of laboratory results by field experimentation; evolution of regulations from the total concentration of a pollutant to its bioavailable fraction; and the use of biomass for added-value products, fine chemicals and biofuels in biorefineries or the production of ornamental plants.

Applying amendments to contaminated soil has been considered a successful strategy to control lead (Pb) pollution. In this study, four different types of amendment (calcium hydrogen phosphate, CHP; hydroxyapatite, HA; ordinary Portland cement, OPC; lime, LI) at two treatment levels were used to immobilize Pb in three contaminated soils. The effectiveness of Pb immobilization was assessed by coupling a sequential extraction procedure (fraction) with in vivo mouse model (Pb relative bioavailability, Pb RBA) and in vitro gastrointestinal assays (bioaccessibility). For all four amendments, Pb RBA generally decreased in YNGJ and HNZZ, with a stronger effect at a high treatment level, but less effective in HNJY. In contrast, when in vitro gastrointestinal simulation tests were used, Pb bioaccessibility determined by SBRC and PBET was generally reduced in most cases, especially in soils treated with phosphate amendments. Sequential extraction procedure demonstrated that the addition of 4 amendments generally decreased the proportion of E1 + C2 compared to untreated soils, while increasing R5, O4, or F3. The relationship between Pb fractions and RBA/bioaccessibility indicated that the bioavailable Pb is primarily from the sum of E1 and C2. The finding of this study highlighted reducing E1 + C2 was a primary strategy to further decrease bioavailable Pb in amended soils, and monitoring Pb fractions may provide a concise and alternative method for comprehending the oral bioavailability of Pb to humans. 

PurposeAnthropic activities induce severe metal(loid)s contamination of many sites, which is a threat to the environment and to public health. Indeed metal(loid)s cannot be degraded, and thus accumulate in soils. Furthermore, they can contaminate surrounding ecosystems through run-off or wind erosion. This study aims to evaluate the phytostabilization capacity of Salix viminalis to remediate As and Pb highly contaminated mine site, in a biochar-assisted phytoremediation context and to assess biochar particle size and dose application effects.Materials and methodsTo achieve this, mesocosm experiments were conducted using the contaminated technosol and four different size fraction of one biochar as amendment, at two application rates (2 and 5%). Non-rooted cuttings of Salix viminalis were planted in the different mixtures. In order to characterize the mixtures, soil pore waters were sampled at the beginning and at the end of the experiment and analyzed for pH, electrical conductivity, and metal(loid) concentrations. After 46 days of Salix growth, roots, stems, and leaves were harvested and weighed, and As and Pb concentrations and distributions were measured.Results and discussionSoil fertility improved (acidity decrease, electrical conductivity increase) following biochar addition, whatever the particle size, and the Pb concentration in soil pore water decreased. Salix viminalis did not grow on the non-amended contaminated soil while the biochar amendment permitted its growth, with a better growth with the finest biochars. The metal(loid)s accumulated preferentially in roots.ConclusionsFine biochar particles allowed S. viminalis growth on the contaminated soil, allowing this species to be used for technosol phytostabilization.

Purpose The possibility of using chemical and microbial additives to enhance the phytoextraction of mercury (Hg) and arsenic (As) from a multi-contaminated soil could be very effective, leading to a significant saving in terms of time and costs of the reclamation. The aim of this study was to evaluate the efficacy of the addition of (i) thiosulfate and (ii) metal-tolerant bacteria isolated from the polluted soil having plant growth promotion (PGP) potential to perform As and Hg phytoextraction by Brassica juncea and Lupinus albus . Materials and methods A collection of 13 bacterial isolates able to tolerate As and Hg was obtained from the contaminated soil, identified by partial 16S rRNA gene sequencing and tested in vitro for PGP activities. The most promising strains were further tested in vivo for the evaluation of plant growth ability and rhizocompetence on model plants. Pot experiments were conducted in microcosms, with polluted soil vegetated with B. juncea and L. albus . Ammonium thiosulfate and potassium dihydrogen phosphate were used as mobilizing agents, together with a bacterial consortium composed by the most promising PGP isolates. Results and discussion Thirteen indigenous metal-tolerant bacterial strains were isolated, and their in vitro characterization highlighted their great potential in assisting the phytoremediation process; most of them tolerated both trace elements and showed, at the same time, multiple PGP traits. The results were confirmed in vivo on model plants and lead to the selection of the most promising PGP strains to be applied in microcosm-scale phytoextraction experiments. Thiosulfate addition significantly increased the mobilization of both elements, promoting bioavailability and phytoextraction. When a selected bacterial consortium was supplemented in addition to thiosulfate, the efficacy of the phytoaccumulation was increased up to 85 % for As and up to 45 % for Hg. Conclusions The use of the common fertilizer thiosulfate appeared to have great potential in phytoextraction practices since it was able to facilitate the uptake by plants of both Hg and As. Moreover, the application of a consortium of indigenous PGP bacteria (PGPB) produced a further positive effect on the plant biomass, supporting and enhancing the phytoextraction strategy, thus demonstrating their potential in a microbe-assisted phytoremediation intervention.

PurposeThis study delineated the effect of calcium (Ca) and ethylenediaminetetraacetic acid (EDTA) addition on arsenic (As) accumulation and physiological attributes of Vicia faba L.Materials and methodsTwo separate experiments were performed. In the first experiment, V. faba seedlings, grown under hydroponic conditions, were exposed to three levels of As (25, 125 and 250 μM) in the presence and absence of three levels of EDTA (25, 125, 250 μM) and calcium (CaCl2: 1, 5 and 10 mM). The effect of EDTA and Ca on As accumulation and physiological attributes of V. faba was assessed by determining As contents in roots and shoot, chlorophyll contents, H2O2 contents, and lipid peroxidation in young and old leaves. In the second experiment, V. faba seeds were grown in As-contaminated sand culture using the same treatment plan.Results and discussionThe accumulation and toxicity of As to V. faba plants increased with increasing As levels in nutrient solution. Arsenic exposure enhanced the production of reactive oxygen species (ROS) in both roots and leaves, which resulted in lipid peroxidation and decreased chlorophyll contents. The presence of both EDTA and Ca, in general, significantly decreased As accumulation by V. faba seedlings, Ca being more effective than EDTA. Both the amendments decreased As-induced reactive oxygen species (ROS) production and lipid peroxidation. In the case of chlorophyll contents, EDTA significantly decreased chlorophyll contents, while Ca significantly increased chlorophyll contents compared to As. The effect of all the treatments was more pronounced in roots than leaves and in young leaves compared to old leaves.ConclusionsIt is proposed that EDTA and Ca greatly affect As accumulation and toxicity to V. faba, and the effect varies greatly with their applied levels as well as type and age of plant organs. The germinating seedlings of V. faba may be preferred for risk assessment studies, while transplanting 1-week-old V. faba seedlings to As-contaminated soils can decrease its toxicity.

PurposeThe subjects of this study were to investigate the remediating potential of the co-cultivation of Pleurotus eryngii and Coprinus comatus on soil that is co-contaminated with heavy metal (cadmium (Cd)) and organic pollutant (endosulfan), and the effects of the co-cultivated mushrooms on soil biochemical indicators, such as laccase enzyme activity and bacterial counts.Materials and methodsA pot experiment was conducted to investigate the combined bioremediation effects on co-contaminated soil. After the mature fruiting bodies were harvested from each pot, the biomass of mushrooms was recorded. In addition, bacterial counts and laccase enzyme activity in soil were determined. The content of Cd in mushrooms and soil was detected by the flame atomic absorption spectrometry (FAAS), and the variations of Cd fractions in soil were determined following the modified BCR sequential extraction procedure. Besides, the residual endosulfan in soil was detected by gas chromatography-mass spectrometry (GC-MS).Results and discussionThe results indicated that co-cultivation of P. eryngii and C. comatus exerted the best remediation effect on the co-contaminated soil. The biomass of mushroom in the co-cultivated group (T group) was 1.57–13.20 and 19.75–56.64% higher than the group individually cultivated with P. eryngii (P group) or C. comatus (C group), respectively. The concentrations of Cd in the fruiting bodies of mushrooms were 1.83–3.06, 1.04–2.28, and 0.67–2.60 mg/kg in T, P, and C groups, respectively. Besides, the removal rates of endosulfan in all treatments exceeded 87%. The best bioremediation effect in T group might be caused by the mutual promotion of these two kinds of mushrooms.ConclusionsThe biomass of mushroom, laccase activity, bacterial counts, and Cd content in mushrooms were significantly enhanced, and the dissipation effect of endosulfan was slightly higher in the co-cultivated group than in the individually cultivated groups. In this study, the effect of co-cultivated macro fungi P. eryngii and C. comatus on the remediation of Cd and endosulfan co-contaminated soil was firstly reported, and the results are important for a better understanding of the co-remediation for co-contaminated soil.

Soil and wastewater samples contaminated by petroleum-related industries were collected from various locations in Saudi Arabia, a country known for its vast oil reserves. The samples were analyzed for their physicochemical properties, including the presence of metals, petroleum hydrocarbons, and aromatic compounds. A total of 264 fungal isolates were analyzed and categorized into eight groups of Aspergillus (194 isolates) and four groups of Penicillium (70 isolates). The potential of these fungal groups to grow in oil or its derivatives was investigated. Two isolates, Aspergillus tubingensis FA-KSU5 and A. niger FU-KSU69, were utilized in two remediation experiments—one targeting wastewater and the other focusing on polluted soil. The FA-KSU5 strain demonstrated complete removal of Fe 3+ , As 3+ , Cr 6+ , Zn 2+ , Mn 2+ , Cu 2+ and Cd 2+ , with bioremediation efficiency for petroleum hydrocarbons in the wastewater from these sites ranging between 90.80 and 98.58%. Additionally, the FU-KSU69 strain achieved up to 100% reduction of Co 2+ , Ba 2+ , B 3+ , V + , Ni 2+ , Pb 2+ and Hg 2+ , with removal efficiency ranging from 93.17 to 96.02% for aromatic hydrocarbons after 180 min of wastewater treatment. After 21 days of soil incubation with Aspergillus tubingensis FA-KSU5, there was a 93.15% to 98.48% reduction in total petroleum hydrocarbons (TPHs) and an 88.11% to 97.31% decrease in polycyclic aromatic hydrocarbons (PAHs). This strain exhibited the highest removal rates for Cd 2+ and As 3+ followed by Fe 3+ , Zn 2+ , Cr 6+ , Se 4+ and Cu 2+ . Aspergillus niger FU-KSU69 achieved a 90.37% to 94.90% reduction in TPHs and a 95.13% to 98.15% decrease in PAHs, with significant removal of Ni 2+ , Pb 2+ and Hg 2+ , followed by Co 2+ , V + , Ba 2+ and B 3+ . The enzymatic activity in the treated soils increased by 1.54- to 3.57-fold compared to the polluted soil. Although the mixture of wastewater and polluted soil exhibited high cytotoxicity against normal human cell lines, following mycoremediation, all treated soils and effluents with the dead fungal biomass showed no toxicity against normal human cell lines at concentrations up to 500 µL/mL, with IC 50 values ≥ 1000 µL/mL. SEM and IR analysis revealed morphological and biochemical alterations in the biomass of A . tubingensis FA-KSU5 and A. niger FA-KSU69 when exposed to petroleum effluents. This study successfully introduces non-toxigenic and environmentally friendly fungal strains play a crucial role in the bioremediation of contaminated environments. Both strains serve as low-cost and effective adsorbents for bio-remediating petroleum wastewater and oil-contaminated soil. Heavy metals and hydrocarbons, the primary pollutants, were either completely removed or reduced to permissible levels according to international guidelines using the dead biomass of FA-KSU5 and FA-KSU69 fungi. Consequently, the environments associated with this globally significant industry are rendered biologically safe, particularly for humans, as evidenced by the absence of cytotoxicity in samples treated with A. tubingensis FA-KSU5 and A. niger FA-KSU69 on various human cell types.

The multiyear cultivation of Miscanthus × giganteus Greef et Deu ( M. × giganteus ) at the soils polluted by metal(loid)s were researched. The biomass parameters and concentrations of elements: Ti, Mn, Fe, Cu, Zn, As, Sr, and Mo were determined in the plant’s organs at harvest. The same metal(loid)s were monitored in the plant’s leaves throughout three vegetation seasons. The principal component analysis and general linear model approaches were applied for statistical evaluation followed by Box-Cox transformation. The difference in the distribution of elements in the plant, the content of elements in the soil, various regime of uptake to the plant tissues, and the year of vegetation were analyzed as driving factors of the phytoremediation. The results showed that the leading promoter was the factor of the zone, which was the most essential for Ti, Fe, and Cu and the smallest for Mn. The factor of differences in soil pollution was essential for Zn and Mo, much less for As, Sr, and Mn, limited for Fe, and was not seen for Ti and Cu. The factor of the interrelation effects of the zone and experiment reflected the different regime of uptake for the plant tissues was seen for two elements: more prominent for Cu and smaller for Ti. While analyzing the dynamic of foliar concentrations of the metal(loid)s during 3 years, two groups were defined. Firstly, Fe, Ni, Mn, and Sr showed stable curves with limited distribution of the plant life cycle. Secondly, As, Zn, Cu, and Mo showed different fluctuations in the curves, which can be attributed to essential influence of those elements to the plant life cycle. Further research will be focused on the application of M. × giganteus to the polluted soil in a bigger scale and comparison results of laboratory and field experiments.

The remediation of mixed contaminated soil is challenging as it often requires actions to minimize metal-induced risks while degrading organic contaminants. Here, the effectiveness of different bioremediation strategies, namely, rhizoremediation with native plant species, mycoremediation with Pleurotus ostreatus spent mushroom substrate, and biostimulation with organic by-products (i.e., composted sewage sludge and spent mushroom substrate), for the recovery of a mixed contaminated soil from an abandoned gravel pit was studied. The combination of biostimulation and rhizoremediation led to the most significant increase in soil health, according to microbial indicator values. The application of composted sewage sludge led to the highest reduction in anthracene and polychlorinated biphenyls concentrations. None of the strategies managed to decrease contamination levels below regulatory limits, but they did enhance soil health. It was concluded that the biological remediation treatments improved soil functioning in a short time, before the concentration of soil contaminants was significantly reduced.

PurposeThe purpose of this paper is to study the responses of soil biological parameters as indicator of ecological status on PAH-contaminated soil.Materials and methodsStudies are conducted on the soils and natural grassy vegetation of monitoring plots subjected to Novocherkassk power station (NPS) emissions. Monitoring plots were established at different distances from the NPS (1.0–20.0 km).Results and discussionThe level of polycyclic aromatic hydrocarbons (PAHs) around NPS is the highest at the monitoring plot located at distance 1.6 km to the northwest through the prevailing wind direction. Gradually, decrease of PAHs was observed while increasing the distance from the NPS through the prevailing wind direction. Calculation of correlations between PAH level and biological activity parameters of soils showed lack of dependence with total and every PAH content in all 12 studied monitoring plots. The most significant correlations were found between PAH content and enzyme activity in the monitoring plots situated through the prevailing wind direction from NPS.ConclusionsThe main pollution source in the studied area is NPS. It was found that contamination of soil by PAHs has a direct dependence on the activity of all biological communities in chernozems, as well as the activities of dehydrogenase and the phytotoxicity of soils. Inverse correlations have been revealed between the PAH contamination and abundance of soil bacteria.